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Biomedical subjects

R Fu

Publications and source records attributed to R Fu.

At least 55 records · Page 3Linked to original sources
PubMed1996

Divergent double chirp pulses for refocusing quadrupolar interactions.

To excite and refocus single quantum coherences in a three-level system with a quadrupolar interaction, it is possible to use frequency-modulated pulses with two diverging frequency components that are swept simultaneously through the two single-quantum transitions. This is achieved by time-dependent amplitude-modulation of the carrier frequency, so as to generate two sidebands with frequencies that are diverging (or converging) symmetrically with respect to the carrier frequency. Simulations and experiments show that such modulated pulses can be used to refocus single quantum coherences to form quadrupole echoes, using much weaker RF amplitudes than in conventional echo sequences. To visualize the evolution of the coherences in a three-level system, a graphical representation of the density operator components is proposed.

Magnetic Resonance Spectroscopy↗
PubMed1995

Increased synovial expression of the adhesion molecules CD66a, CD66b, and CD31 in rheumatoid and osteoarthritis.

Leukocyte-endothelial interaction mediated by adhesion molecules may play a role in the ingress of inflammatory cells into the rheumatoid (RA) synovial tissue (ST). A number of these molecules have been shown to be up-regulated in the inflamed compared to normal ST. We studied the distribution of two members of the CD66 carcinoembryonic antigen adhesion molecule family, as well as that of CD31, an antigen structurally related to CD66, on various cell types in the RA compared to osteoarthritic (OA) and normal ST. Immunoperoxidase histochemistry was carried out using monoclonal antibodies to CD66a, CD66b, and CD31. This study was performed on ST from 10 patients with RA, 10 with OA, and 4 normal individuals. CD66a, and CD66b were expressed on RA and OA ST myeloid cells but not on normal ST lining cells and interstitial macrophages, suggesting that these antigens may be specific markers of diseased compared to normal ST macrophages (P < 0.05). CD31 was present on more RA and OA than on normal ST macrophages. Also, CD31 was present on most RA, OA, and normal ST endothelial cells. Our results indicate that the expression of CD66a, CD66b, and CD31, members of the immunoglobulin superfamily of adhesion receptors, is up-regulated on cells of myeloid origin in the inflamed compared to normal ST. These results suggest that the CD66 antigens and CD31 may be involved in the adhesive events in the inflamed synovium.

Antigens, CD↗
PubMed1995

Increased synovial expression of transforming growth factor (TGF)-beta receptor endoglin and TGF-beta 1 in rheumatoid arthritis: possible interactions in the pathogenesis of the disease.

The ingress of inflammatory cells into the rheumatoid (RA) synovial tissue (ST) plays a role in the pathogenesis of this disease. Transforming growth factor beta (TGF-beta) may play a role in this process. We have investigated the distribution of endoglin, a newly described receptor for TGF-beta 1 and -beta 3, in RA compared to osteoarthritis (OA) or normal ST. Immunohistochemical analysis was carried out using an anti-TGF-beta 1 monoclonal antibody (mAb) as well as 10 mAbs raised against various epitopes of endoglin. This study was performed on ST from 10 patients with RA, 10 with OA, and 4 normal individuals. TGF-beta 1 expression was significantly up-regulated on RA compared to OA and normal ST lining cells, interstitial macrophages, and endothelial cells (P < 0.05). All anti-endoglin mAbs uniformly reacted with endothelial cells in RA, OA, and normal STs. However, 3 out of 10 anti-endoglin mAbs reacted with significantly more RA versus normal ST lining cells (P < 0.05), as well as RA compared to OA and normal macrophages (P < 0.05). There was a positive correlation between TGF-beta 1 and endoglin reactivity on the synovial lining layer and subsynovial macrophages (P < 0.05). These results indicate that TGF-beta 1 and certain epitopes of endoglin, a TGF-beta 1 and -beta 3 receptor, are up-regulated on myeloid elements in RA compared to normal ST. Endoglin is also present on ST endothelia, and its expression may also be increased on OA compared to normal ST lining cells. These findings implicate endoglin in the pathogenesis of RA.

Antibodies, Monoclonal↗
PubMed1995

Macrophage inflammatory protein-1 beta: a C-C chemokine in osteoarthritis.

The aim of this study was to determine whether the cytokine macrophage inflammatory protein-1 beta (MIP-1 beta) is present and functionally active in the arthritic joint. We used immunoassays and bioassays to assess the presence and function of MIP-1 beta using samples obtained from 62 arthritic patients. MIP-1 beta levels were increased in synovial fluids (SFs) from patients with osteoarthritis (OA) (18.0 +/- 8.9 ng/ml) (SD) compared to patients with rheumatoid arthritis (RA) 6.1 +/- 2.9 ng/ml) or other forms of arthritis (10.4 +/- 7.0 ng/ml) (P < 0.05). Levels of OA SF MIP-1 beta were significantly greater than OA or normal serum levels of MIP-1 beta. Anti-MIP-1 beta neutralized 28% of the chemotactic activity for monocytes found in OA SFs. Isolated OA synovial tissue fibroblasts did not constitutively produce MIP-1 beta but could be induced to express this chemokine upon exposure to tumor necrosis factor-alpha, interleukin-1 beta, or lipopolysaccharide. Synovial tissue immunohistochemical staining revealed that the main immunopositive cells in OA were the lining cells as well as vascular smooth muscle and endothelial cells. A minority of macrophages were immunopositive as well. In this study, we identify MIP-1 beta as a unique cytokine increased in OA compared to RA SF. We conclude that MIP-1 beta may play a role in the ingress of monocytes into the OA joint.

Arthritis↗
PubMed1995

[A 3-year follow-up study of 50 children with acute lymphoblastic leukemia].

Fifty children with acute lymphoblastic leukemia were followed up for an average period of 3 3/12 years. The results showed that 19 (38%) cases were given up after diagnosis. Eight patients only received treatment irregularly for 1 year; among them, 2 discontinued therapy by themselves for 2-3 years and survived free of events, and 6 were lost, Twenty-three received treatment regularly; among them, 3 died of infection, 11 were in continuously complete remission and 9 experienced relapse. The total disease-free survival rate for mean 3 3/12 years is 42% (13/31), exclusive of the 19 given up cases. Statistical analysis indicates a higher relapse rate in those cases with lower initial glucocorticoid receptors (GCR) of peripheral blood lymphocytes.

Child↗
PubMed1995

Demonstration of in vitro cultured exoerythrocytic schizonts and hypnozoites of Plasmodium vivax (southern China isolate) by an immunoperoxidase antibody technique.

By indirect immunoperoxidase staining, different forms of exoerythrocytic (EE) stage of Plasmodium vivax (Southern China isolates) are revealed in d8 cultured material. The mature schizonts are elongated in shape measuring 42-48 microns in diameter, immature schizonts 14-28 microns and hypnozoites 4-7 microns. EE schizonts are stained dark-brown only after conjugated by monoclonal antibody (McAb) 4B2 specific against erythrocytic stages of P. vivax while hypnozoites are only stained after conjugated by McAb 2F2 against sporozoite. These results show that the antigenic components of these two forms of EE plasmodia are quite different. The ratio of EE schizont and hypnozoite found within hepatoma cells (HepG2-A16) is 1.5 to 1. Referred to the clinical manifestations of the isolate, among 5 volunteers not radically cured, two had long incubation period (283 d and 304 d, respectively) and three relapsed 235, 260 and 365 days after the primary attack. These data are unanimous with the comparatively large ratio of hypnozoites in the cultured material.

Animals↗
PubMed1995

[Research on the factors influencing the sporogonic multiplication of Plasmodium vivax in the mosquito vector].

Volunteers with no previous malaria history infected with northern Guangdong isolate of Plasmodium vivax and mosquito vector, Anopheles stephensi, were the objects and materials used in the research. Totally there were 16 volunteers, half of them were infected by mosquito-bite and another half by blood-inoculation. Blood was drawn when parasitemia reached a certain level and the erythrocytic forms developed into large trophozoites. Mosquitoes fed on infected blood from 5 and 4 donors of the two groups gave high gland infection rate and high gland indices respectively, but the highest gland indices were got from volunteers infected by mosquito-bite. The time of blood-feeding is important, only those blood samples taken from the people 3 to 8 days after the first onset gave good sporozoite harvest and the gland infection was greatly reduced on the 9th day. During this period (d3-8), the density of gametocyte had obvious influence on the intensity of mosquito infection. When the sex ratio of gametocyte (female:male) was 4:1 or less, the result would be good. For the purpose of reflecting more perfectly the circumstance of the sporogonic multiplication of plasmodium in the mosquito vector, we suggest the concept of gut/gland infection intensity which equals the product of the infection rate and the index of oocyst/positive gland.

Adult↗
PubMed1995

[Effect of environmental temperature, cryopreservation and aging on Plasmodium vivax sporozoites developing into exoerythrocytic stages].

After Anopheles stephensi mosquitoes with salivary infection of Plasmodium vivax were put in environments with temperatures of 30 +/- 1 degrees C, 26 +/- 1 degrees C or 13 +/- 1 degrees C for 5 d, their glands were aseptically dissected and sporozoites were collected and inoculated into HepG2-A16 cell monolayers. Seven days post-inoculation the cultured materials were harvested and the exoerythrocytic schizonts and hypnozoites were observed under the microscope by using immunoperoxidase staining technique. The results showed that the sporozoite developing rate of 30 +/- 1 degrees C group and 13 +/- 1 degrees C group was significantly lower than that of 26 +/- 1 degrees C group (0.33%, 0.35% and 0.75% respectively). The proportion of hypnozoites in the total number of EE forms was the highest in the low temperature group (62.5%) compared with 26 +/- 1 degrees C and 30 +/- 1 degrees C group (40.1% and 42.7% respectively). Suggesting that the low environmental temperature first affected the viability of tachysporozoites or the phenotype of sporozoites and thus resulted in heightened hypnozoite rate. This is parallel to the epidemiological data that in the regions of high latitute vivax malaria with long incubation period was more frequently observed. When the sporozoites within the body of mosquito were cryopreserved at -70 degrees C or in liquid nitrogen for 24 h or 5 d respectively, the proportion of hypnozoite increased 87.4% and 82.4%, respectively. However, cryopreservation did not inactivate all of the tachysporozoites, indicating that the resistance to ultralow temperature in bradysporozoite was much greater than that in tachysporozoites. Aging of sporozoites decreased their developing rate and the exoerythrocytic (EE) schizonts were found to grow sluggishly and asynchronously, indicating that the size of EE schizont and the age of sporozoites are in negative correlation. Meantime, proportion of the hyponozoite decreased significantly.

Aging↗
PubMed1995

[Observation on hypnozoite of different isolates of Plasmodium vivax in cultured materials].

The hypnozoites of different isolates of P. vivax from Shenzhen and Shixin, Guangdong Province, Changsha, Hunan Province and Yincheng, Hubei Province of China in cultured materials were observed employing immunoperoxidase staining method. The percentages of hypnozoites among exoerythrocytic stages were 40.1, 43.5, 50.6 and 57.1%, respectively, indicating that the percentage of hypnozoites and the latitude are in positive correlation. The equation is y = -6.68 + 2.05x. When the difference in latitude of Plasmodium vivax isolate source was more than 5 degrees, a significant difference in the percentage of hypnozoites in various geographic isolates was found. However, the average diameters of hypnozoites and schizonts among various geographic isolates showed no significant difference. The results showed that in the regions north to the Yangtze River, the proportion of hypnozoite in the liver stage of P. vivax was larger than those in the southern China, being consistent with the clinical manifestations.

Animals↗
PubMed1995

Effect of monoclonal antibodies on the entry and development of Plasmodium vivax sporozoite in cultured cells.

We have observed the effect of monoclonal antibodies against sporozoites (2F2,NV3,2E10) and blood stages (4B2,8E3) of P. vivax on entry and subsequent development of P. vivax sporozoites in HepG2--A16 cells in vitro. The results demonstrated that inhibitory effects of monoclonal antibodies on the attachment and entry were found to be related to the antibody concentration. At 25 micrograms/ml, the percentages of inhibition of sporozoite invasion were 100% (2F2), 76%(NVS3) and 10.5% (2E10). Even if the invasion was not totally inhibited, the presence of abnormal exoerythrocytic schizonts suggested that continued effect of antibodies after sporozoite penetration still existed. No significant effects of 4B2 (4.5%) and 8E3 (3.4%) were recorded as compared with normal mouse serum. These findings indicate that inhibition of sporozoite invasion assay may be useful for determining the protective effect of anti-sporozoite antibodies in vitro.

Animals↗
PubMed1994

DcrA, a c-type heme-containing methyl-accepting protein from Desulfovibrio vulgaris Hildenborough, senses the oxygen concentration or redox potential of the environment.

The amino acid sequence of DcrA from Desulfovibrio vulgaris Hildenborough, a strictly anaerobic, sulfate-reducing bacterium, indicated homology with the methyl-accepting chemotaxis proteins from enteric bacteria (A. Dolla, R. Fu, M. J. Brumlik, and G. Voordouw, J. Bacteriol. 174:1726-1733, 1992). The homology is restricted to the cytoplasmic C-terminal signaling domain. The periplasmic N-terminal sensor domain was found to contain a unique sequence, CHHCH, corresponding to a consensus c-type heme binding site. A pretreated, DcrA-specific polyclonal antiserum, generated against DcrA protein overproduced in Escherichia coli, was used for immunoprecipitation of 35S-labeled DcrA from D. vulgaris and Desulfovibrio desulfuricans G200(pJRFR2), a transconjugant that overexpresses functional DcrA. Labeling of the latter with the heme precursor 5-amino-[4-14C]levulinic acid, followed by immunoprecipitation, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and fluorography, confirmed the presence of c-type heme, while labeling with L-[methyl-3H]methionine in the absence of protein synthesis confirmed that DcrA is a methyl-accepting protein. The base liability of the incorporated radioactivity indicated methyl ester formation like that occurring in the methyl-accepting chemotaxis proteins of enteric bacteria. L-[methyl-3H]methionine labeling of D. desulfuricans G200(pJRFR2) under different conditions indicated that methyl labeling of DcrA decreased upon addition of oxygen and increased upon subsequent addition of the reducing agent dithionite. These results indicate that DcrA may serve as a sensor of oxygen concentration and/or redox potential.

Amino Acid Sequence↗
PubMed1994

[In vitro cultivation of the exoerythrocytic stage of Plasmodium vivax (southern China isolate)].

An in vitro culture system for the exoerythrocytic (EE) stage of Plasmodium vivax was first developed in our laboratory in China. Anopheles stephensi mosquitoes were infected by membrane feeding with heparinized blood from a volunteer. After 14-18 days, the mosquito salivary glands were aseptically dissected in culture medium and ground in a tissue grinder to form sporozoite suspension. Sporozoites were counted and added to the cultures of monolayer hepatoma cells at the number of 4.9 x 10(4)-1.3 x 10(5) per cover glass. Sometimes, 8-10 pairs of infected gland were added directly to the cultured cells. On day 7, EE schizonts of P. vivax were found in cultures. In addition, normal erythrocytes (type o) were added to the cultures on day 10 at a concentration of 10(8) per dish. Fourteen days later, erythrocytes in culture supernatant were collected and thin blood films were made. Numerous intra-erythrocytic P. vivax parasites were identified on the films after Giemsa staining. Most intra-erythrocytic forms were rings and early large trophozoites. Schuffer's dots were present in many of the infected cells which were pale and obviously enlarged. These results indicated that the in vitro hepatic cycle of P. vivax was established.

Animals↗
PubMed1993

The hmc operon of Desulfovibrio vulgaris subsp. vulgaris Hildenborough encodes a potential transmembrane redox protein complex.

The nucleotide sequence of the hmc operon from Desulfovibrio vulgaris subsp. vulgaris Hildenborough indicated the presence of eight open reading frames, encoding proteins Orf1 to Orf6, Rrf1, and Rrf2. Orf1 is the periplasmic, high-molecular-weight cytochrome (Hmc) containing 16 c-type hemes and described before (W. B. R. Pollock, M. Loutfi, M. Bruschi, B. J. Rapp-Giles, J. D. Wall, and G. Voordouw, J. Bacteriol. 173:220-228, 1991). Orf2 is a transmembrane redox protein with four iron-sulfur clusters, as indicated by its similarity to DmsB from Escherichia coli. Orf3, Orf4, and Orf5 are all highly hydrophobic, integral membrane proteins with similarities to subunits of NADH dehydrogenase or cytochrome c reductase. Orf6 is a cytoplasmic redox protein containing two iron-sulfur clusters, as indicated by its similarity to the ferredoxin domain of [Fe] hydrogenase from Desulfovibrio species. Rrf1 belongs to the family of response regulator proteins, while the function of Rrf2 cannot be derived from the gene sequence. The expression of individual genes in E. coli with the T7 system confirmed the open reading frames for Orf2, Orf6, and Rrf1. Deletion of 0.4 kb upstream from orf1 abolished the expression of Hmc in D. desulfuricans G200, indicating this region to contain the hmc operon promoter. The expression of two truncated hmc genes in D. desulfuricans G200 resulted in stable periplasmic c-type cytochromes, confirming the domain structure of Hmc. We propose that Hmc and Orf2 to Orf6 form a transmembrane protein complex that allows electron flow from the periplasmic hydrogenases to the cytoplasmic enzymes that catalyze the reduction of sulfate. The domain structure of Hmc may be required to allow interaction with multiple hydrogenases.

Amino Acid Sequence↗
PubMed1993

Blood glutathione status during exercise: effect of carbohydrate supplementation.

Blood glutathione status and activities of antioxidant enzymes have been investigated during prolonged exercise with or without carbohydrate (CHO) supplementation. Eight subjects cycled at approximately 70% of maximal oxygen uptake to fatigue [134 +/- 19 (SE) min] on the first occasion (control, CON) and at the same work load and duration on the second occasion but with CHO ingestion during exercise. Blood reduced glutathione (GSH) concentration increased from 0.55 +/- 0.05 mM at rest to 0.77 +/- 0.09 mM after 120 min of exercise during CON (P < 0.01) but remained constant during CHO exercise. Blood glutathione disulfide (GSSG) levels were unchanged during CON and CHO exercise. Blood GSH + GSSG content and GSH/GSSG ratio were also significantly (P < 0.05) elevated during CON but not during CHO exercise. The increases in GSH and GSH + GSSG in CON were associated with decreases in plasma glucose and insulin levels. Activities of blood GSH peroxidase, GSSG reductase, and glucose-6-phosphate dehydrogenase were significantly increased during the CHO exercise, whereas only GSSG reductase activity was elevated during the CON ride. It is concluded that blood GSH increases during prolonged exercise and that CHO supplementation may prevent blood GSH increase possibly because of its inhibitory effects on hepatic hormonal releases, which stimulate GSH output.

Adult↗
PubMed1993

Opioid antagonist diprenorphine microinjected into parabrachial nucleus selectively inhibits vasopressin response to hypovolemic stimuli in the rat.

Subcutaneous injection of the potent, nonselective opioid antagonist diprenorphine inhibits the vasopressin response to acute hypovolemia. To determine if this inhibition is due to antagonism of opioid receptors in brain pathways that mediate volume control, we determined the vasopressin response to different stimuli when diprenorphine or other opiates were injected into the cerebral ventricles, the nucleus tractus solitarius (NTS), or the lateral parabrachial nucleus (PBN) of rats. We found that the vasopressin response to hypovolemia was inhibited by injection of diprenorphine into the cerebral ventricles at a dose too low to be effective when given subcutaneously. This response also was inhibited when a 20-fold lower dose of diprenorphine was injected into the PBN but not when it was injected into the NTS. The inhibitory effect of diprenorphine in the PBN was not attributable to a decrease in osmotic or hypovolemic stimulation and did not occur with osmotic or hypotensive stimuli. Injecting the PBN with equimolar doses of the mu antagonist naloxone, the delta antagonist ICI-154,129 or the kappa-1 agonist U-50,488H had no effect on basal or volume-stimulated vasopressin. We conclude that the inhibition of vasopressin by diprenorphine is due partially to action at a novel class of opioid receptors that transmit volume stimuli through the PBN.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗
PubMed1992

Nucleotide sequence of dcrA, a Desulfovibrio vulgaris Hildenborough chemoreceptor gene, and its expression in Escherichia coli.

The amino acid sequence of DcrA (Mr = 73,000), deduced from the nucleotide sequence of the dcrA gene from the anaerobic, sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough, indicates a structure similar to the methyl-accepting chemotaxis proteins from Escherichia coli, including a periplasmic NH2-terminal domain (Mr = 20,700) separated from the cytoplasmic COOH-terminal domain (Mr = 50,300) by a hydrophobic, membrane-spanning sequence of 20 amino acid residues. The sequence homology of DcrA and these methyl-accepting chemotaxis proteins is limited to the COOH-terminal domain. Analysis of dcrA-lacZ fusions in E. coli by Western blotting (immunoblotting) and activity measurements indicated a low-level synthesis of a membrane-bound fusion protein of the expected size (Mr = approximately 137,000). Expression of the dcrA gene under the control of the Desulfovibrio cytochrome c3 gene promoter and ribosome binding site allowed the identification of both full-length DcrA and its NH2-terminal domain in E. coli maxicells.

Amino Acid Sequence↗
PubMed1992

Responses of glutathione system and antioxidant enzymes to exhaustive exercise and hydroperoxide.

Glutathione (gamma-glutamylcysteinylglycine) is one of the major antioxidants in the body. The present study investigated the changes of glutathione status, oxidative injury, and antioxidant enzyme systems after an exhaustive bout of treadmill running and/or hydroperoxide injection in male Sprague-Dawley rats. Concentrations of total and reduced glutathione in deep vastus lateralis muscle were significantly increased (P less than 0.01) after exhaustive exercise with either hydroperoxide (t-butyl hydroperoxide) or saline injection, whereas hydroperoxide alone had no significant effect. Exhaustive exercise increased muscle glutathione disulfide content by 75 and 60% (P less than 0.05), respectively, in hydroperoxide and saline groups. Concentrations of glutathione-related amino acids glutamate, cysteine, and aspartate were significantly increased in the same muscle after exhaustion. Hepatic glutathione status was not affected by either hydroperoxide injection or exercise. Glutathione peroxidase, glutathione reductase, superoxide dismutase, and catalase activities were significantly elevated after exhaustive exercise with or without hydroperoxide injection in muscle but not in liver. Hydroperoxide and exhaustive exercise enhanced lipid peroxidation in muscle and liver, respectively. It is concluded that exhaustive exercise can impose a severe oxidative stress on skeletal muscle and that glutathione systems as well as antioxidant enzymes are important in coping with free radical-mediated muscle injury.

Amino Acids↗