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Biomedical subjects

T Xu

Publications and source records attributed to T Xu.

At least 19 recordsLinked to original sources
PubMed1996

Transfer of a gene encoding the anticandidal protein histatin 3 to salivary glands.

Mucosal candidiasis, the most common opportunistic fungal infection in human immunodeficiency virus (HIV)-infected patients, is an early sign of clinically overt acquired immunodeficiency syndrome (AIDS) and an important cause of morbidity, particularly in HIV-infected children. The appearance of azole-resistant strains of Candida albicans had made clinical management of candidiasis increasingly difficult. We propose a novel approach to the management of candidal infections that involves the use of naturally occurring antifungal proteins, such as the histatins. Histatins are a family of small proteins that are secreted in human saliva. We have constructed recombinant adenovirus vectors that contain the histatin 3 cDNA. These vectors are capable of directing the expression of histatin 3 in the saliva of rats at up to 1,045 micrograms/ml, well above the levels found in normal human saliva. The adenovirus-directed histatin demonstrated a 90% candidacidal effect in the timed-kill assay against both fluconazole-susceptible and fluconazole-resistant strains of C. albicans and inhibited germination by 45% in the same strains. These studies suggest that a gene transfer approach to overexpress naturally occurring antifungal proteins may be useful in the management of mucosal candidiasis.

Adenoviridae↗
PubMed1996

The effect of mild trypsin digestion of F1 on energy coupling in the mitochondrial ATP synthase.

Mild trypsin digestion of isolated bovine-heart mitochondrial F1-ATPase removed the first 15 residues from the N-terminus of subunit alpha under conditions in which other F1 subunits were apparently untouched. When the trypsinized F1 (TF1) was reconstituted with the F0 sector in the mitochondrial membrane (USMP), the ATP hydrolase activity acquired oligomycin sensitivity but ATP hydrolysis was decoupled from proton pumping. TF1 added to USMP did not block the proton channel in F0 as the native F1 did. AMP-PNP inhibited proton conductivity in reconstituted F1-USMP but this effect was lost in reconstituted TF1-USMP. These results indicate that the N-terminus of the F1 alpha subunit plays a critical role in the conformational communication between F1 and F0.

Adenosine Triphosphate↗
PubMed1996

Candidacidal activity of human salivary histatin recombinant variants produced by site-directed mutagenesis.

Histatin 5 (Hst5) is a 24-amino acid (aa) member of the Hst family that is found in human salivary secretions and exhibits candidacidal activity. Hst5 contains a 13-aa region that alone is capable of killing fungal pathogens and is referred to as the functional domain. To investigate the role of specific aa located within the functional domain, the pRSET bacterial expression system was used to produce recombinant Hst5 (re-Hst5) and several re-variants that were generated by site-directed mutagenesis. The vector pRSETC expresses genes of interest as fusion proteins attached to the carboxy end of an N-terminal His6 tag that binds to nickel (Ni2+). The re-variants were generated using the polymerase chain reaction (PCR) and had Gly substituted for either the His, Glu or Lys/Arg within the functional domain. PCR products that encoded either the wild-type or variant forms of re-Hst5 were inserted into pRSETC and produced as fusion proteins which were affinity purified from cell lysates by Ni(2+)-Sepharose chromatography. Fusion proteins were digested with CNBr and re-Hsts were purified by reversed-phase high performance liquid chromatography (RP-HPLC). Re-Hsts were tested in bioassays to measure the ability to kill both Candida albicans (C. albicans) blastoconidia and spheroplasts which were generated by removal of the cell wall. In both assays, re-Hst5 displayed dose-dependent candidacidal activity that was nearly identical to that of native Hst5 purified from human salivary secretions. Re-Hst5 variants with either Glu or Lys/Arg substitutions demonstrated significantly lower candidacidal activity in both assays, while the variant with His mutated showed essentially no activity at physiological concentrations. These results indicate that acidic and basic aa within the functional domain contribute to candidacidal activity and that the His are essential for candidacidal activity. Additionally, since C. albicans spheroplasts were also susceptible to Hsts, the cell wall is not an essential component in the Hst mechanism of candidacidal action.

Amino Acid Sequence↗
PubMed1996

KUZ, a conserved metalloprotease-disintegrin protein with two roles in Drosophila neurogenesis.

During neurogenesis in Drosophila both neurons and nonneuronal cells are produced from a population of initially equivalent cells. The kuzbanian (kuz) gene described here is essential for the partitioning of neural and nonneuronal cells during development of both the central and peripheral nervous systems in Drosophila. Mosaic analyses indicated that kuz is required for cells to receive signals inhibiting the neural fate. These analyses further revealed that the development of a neuron requires a kuz-mediated positive signal from neighboring cells. The kuz gene encodes a metalloprotease-disintegrin protein with a highly conserved bovine homolog, raising the possibility that kuz homologs may act in similar processes during mammalian neurogenesis.

Amino Acid Sequence↗
PubMed1996

Cholecystokinin octapeptide reverses the kappa-opioid-receptor-mediated depression of calcium current in rat dorsal root ganglion neurons.

Although the cholecystokinin octapeptide (CCK-8) is reported to antagonize the kappa-opioid-receptor-mediated analgesic effect in spinal cord, its mechanism and sites of action remain obscure. In the present study, the whole-cell patch-clamp recording technique was employed to examine the effect of kappa-opioid agonist U50488H on voltage-gated calcium channels and the interaction between the CCK-8 and U50488H in acutely isolated rat dorsal root ganglion neurons. The results indicate that the calcium currents elicited in dorsal root ganglion neurons can be depressed by U50488H, an effect readily reversed by the kappa-opioid receptor antagonist Nor-BNI or by the antiopioid peptide CCK-8. The effect of the CCK-8 can be abolished by the CCK-B receptor antagonist, L365,260. While CCK-8 showed a potent opioid-reversal effect, it by itself exerted a slight inhibitory effect on calcium current. This novel observation in the dorsal root ganglion neurons indicates that CCK-8 can antagonize the kappa-opioid-receptor-mediated depressant effect on voltage-gated calcium current, and this antagonizing effect appears to be mediated via CCK-B receptor.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗
PubMed1996

Reducing frailty and falls in older persons: an investigation of Tai Chi and computerized balance training. Atlanta FICSIT Group. Frailty and Injuries: Cooperative Studies of Intervention Techniques.

OBJECTIVE: To evaluate the effects of two exercise approaches, Tai Chi (TC) and computerized balance training (BT), on specified primary outcomes (biomedical, functional, and psychosocial indicators of frailty) and secondary outcomes (occurrence of falls). DESIGN: The Atlanta FICSIT (Frailty and Injuries: Cooperative Studies of Intervention Techniques), a prospective, randomized, controlled clinical trial with three arms (TC, BT, and education [ED]. Intervention length was 15 weeks, with primary outcomes measured before and after intervention and at 4-month follow-up. Falls were monitored continuously throughout the study. SETTING: Persons aged 70 and older living in the community. PARTICIPANTS: A total of 200 participants, 162 women and 38 men; mean age was 76.2. MEASUREMENTS: Biomedical (strength, flexibility, cardiovascular endurance, body composition), functional (IADL), and psychosocial well-being (CES-D scale, fear of falling questionnaire, self-perception of present and future health, mastery index, perceived quality of sleep, and intrusiveness) variables. RESULTS: Grip strength declined in all groups, and lower extremity range of motion showed limited but statistically significant changes. Lowered blood pressure before and after a 12-minute walk was seen following TC participation. Fear of falling responses and intrusiveness responses were reduced after the TC intervention compared with the ED group (P = .046 and P = .058, respectively). After adjusting for fall risk factors, TC was found to reduce the risk of multiple falls by 47.5%. CONCLUSIONS: A moderate TC intervention can impact favorably on defined biomedical and psychosocial indices of frailty. This intervention can also have favorable effects upon the occurrence of falls. Tai Chi warrants further study as an exercise treatment to improve the health of older people.

Accidental Falls↗
PubMed1996

ATP-sensitive potassium channels regulate stimulated ANF secretion in isolated rat heart.

Perfused hearts (n = 127) were exposed to acute hypoxia (10% O2 for 12 or 20 min) or left atrial stretch (increase in atrial pressure) in the presence or absence of 100 mumol/l ATP-sensitive potassium channel blocker (tolbutamide) or openers (pinacidil and diazoxide). Hypoxia alone elicited a prolonged atrial natriuretic factor (ANF) release, peaking at 74% over baseline (P < 0.01); with tolbutamide, ANF secretion peaked at 132% over baseline (P < 0.01). Pinacidil and diazoxide abolished the ANF response to hypoxia (P < 0.01). Atrial stretch alone (1 mmHg) transiently (2 min) increased ANF by 56% (P < 0.05); with tolbutamide, ANF increased transiently by 124% and showed a prolonged increase of 52% (P < 0.05). With tolbutamide, graded stretch (0.5-2.3 mmHg) induced a bell-shaped transient (2-min) increase of ANF release [-3% at 0.5 mmHg, 124% (P < 0.05) at 1.0 mmHg, 80% (P < 0.05) at 1.48 mmHg, and 14% at 2.22 mmHg] and a saturating prolonged ANF response. Tolbutamide increased the ANF response nonsignificantly at lower doses (30 mumol/l) and had no effect at 1 mumol/l. Pinacidil abolished the stretch-induced ANF release. These results suggest that ATP-sensitive potassium channels are extremely potent modulators of stimulated ANF secretion.

Adenosine Triphosphate↗
PubMed1995

The 10-Hz rhythm in the sympathetic nerve activity of cats, rats and rabbits.

Since the 10-Hz rhythmic activity in the sympathetic nerves was reported only in cats, we examined whether the activity of the same frequency could be observed in rats and rabbits as well as in the cats. Histograms of inter-burst-peak intervals of discharges of the renal nerves revealed that the 100 ms interval activity, the reverse of the 10-Hz, was observed in all the three mammals, of which the baroreceptor afferents were intact or inactivated. Further, the same frequency activity could be evoked by intermittent electrical stimulation of the dorsolateral funiculus of the cervical cord in spinal animals. It was suggested that the 10-Hz rhythmic activity in the sympathetic nerves was a common phenomenon throughout mammals and this activity was produced in the spinal cord. The physiological significance of the 10-Hz activity in the sympathetic nerves was discussed.

Afferent Pathways↗
PubMed1995

Recombinant histatins: functional domain duplication enhances candidacidal activity.

Histatin 3 (Hst3) is a 32-amino-acid (aa) His-rich protein with antimicrobial activity found in human salivary secretions. To explore further the structure/function relationship of Hst, we utilized a bacterial system for the efficient production of recombinant Hst3 (re-Hst3) and Hst variants. Previously, we demonstrated that the middle portion of Hst3 (aa 13-24) contains the functional domain responsible for killing Candida albicans. Using PCR and splice overlap extension, a Hst variant (re-Hst3rep) was made in which the functional domain was repeated in tandem. Using the pRSET bacterial expression system, re-Hst3 and the variant re-Hst3rep were produced as chimeric fusions and were isolated from bacterial sonicates by affinity chromatography. Affinity purified fusion proteins were digested with CNBr and re-Hst were separated from their fusion partners by reverse-phase high-performance liquid chromatography. The activity of re-Hst3 and re-Hst3rep was compared to that of native Hst3 from human salivary secretions in the C. albicans killing assay. The LD50 values for candidacidal activity of native Hst3, re-Hst3 and re-Hst3rep were 7.2, 6.8 and 4.1 nmol/ml, respectively. At lower concentrations re-Hst3rep was five times more active than native Hst3 or re-Hst3 and at even lower concentrations re-Hst3rep exhibited significant candidacidal activity while native Hst3 and re-Hst3 were inactive. These results demonstrate an expression system for production of biologically active functional Hst and Hst variants and shows that repetition of the functional domain of Hst3 enhances candidacidal activity.

Amino Acid Sequence↗
PubMed1995

The spinally mediated 10-Hz rhythm in the sympathetic nerve activity of cats.

To examine the origin of the so-called '10-Hz rhythm' in the sympathetic nerves, the mass discharges of the white ramus of the third thoracic segment (T3WR) and the inferior cardiac nerve (ICN) and the activities of single postganglionic neurons in the stellate ganglion were recorded in spinal cats. During the chemical or electrical stimulation of the spinal cord, the time of peak of discharges in the sympathetic nerves was analyzed. Both intrathecal administration of N-methyl-D-aspartic acid (NMDA; 3-10 mM) and continuous high frequency (80-200 Hz) electrical stimulation of the dorsolateral funiculus at the second cervical level increased activity of the sympathetic nerves in a similar fashion. In these conditions, modes of the inter-peak interval histograms (IPIH) were about 100 ms (range; 90-130 ms), the inverse of about 10 Hz, but no correlation was observed in autocorrelograms of these peaks of discharges. Therefore, this 100-ms interval activity might have some significance for the 10-Hz rhythm. In order to make this point clear, we stimulated the dorsolateral funiculus with intermittent trains of electrical pulses (0.2-ms duration, 10-35 pulses of 80-200 Hz frequency, in every 300-800 ms). While intermittent trains of pulses were applied, multiple peaks of discharges were evoked in the sympathetic nerves. IPIHs of the nerves were multimodal. The first mode (shortest interval) was about 100 ms. The first mode depended on none of the stimulus parameters but the probability of the about 100-ms interval activity depended on the interval of trains of pulses and the stimulus intensity. With this intermittent stimulation, the autocorrelogram of the peaks revealed the 100-ms interval rhythm. To confirm that the peak of discharges in the ICN was composed of synchronized spikes of postganglionic fibers, single neuronal activities of postganglionic neurons were recorded during the intermittent stimulation. Inter-spike interval histograms showed almost same profile as the IPIHs of the ICN. These results can be explained if the following two assumptions are valid; (i) There are mechanisms that limit minimum firing interval of most preganglionic neurons to about 100 ms. (ii) Simultaneously a interneuron in the spinal cord resets the spike generation of multiple preganglionic neurons. Similarity of the spike activities of the sympatho-excitatory reticulospinal neurons to the intermittent stimulation can explain the 10-Hz rhythm in the peripheral sympathetic nerves in intact spinal cord animals. It is not necessary to postulate the specific 10-Hz rhythm generator in the brain stem for the sympathetic nervous system.

Action Potentials↗
PubMed1995

Functional comparison of native and recombinant human salivary histatin 1.

Histatin 1 is a histidine-rich phosphoprotein present in human parotid saliva that possesses candidacidal activity and functions in mineralization by adsorbing to hydroxyapatite. The objective of the present study was to develop a system for recombinant production of histatin 1 and to examine the role of phosphorylation in the functional activities of this molecule. Native histatin 1 (containing a phosphoserine at residue 2) was purified from parotid saliva, whereas a bacterial expression system was used to produce a recombinant form of histatin 1 (re-Hst1) that lacked phosphorylated serine. Histatin 1 cDNA was inserted into the vector pGEX-3X, which expresses foreign genes as soluble fusion proteins attached to the carboxyl-terminus of glutathione S-transferase (GST). The GST/re-Hst1 fusion protein was isolated from cell lysates by affinity chromatography on glutathione (GSH)-Sepharose and digested with cyanogen bromide to separate re-Hst1 from the GST fusion partner. The digest was subjected to reversed-phase high-performance liquid chromatography on a C18 column, and re-Hst1 was eluted as a well-defined peak. The yield of re-Hst1 was 4 mg/L of bacterial culture. Amino-terminal sequencing and amino acid analysis confirmed the final product as re-Hst1. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that native histatin 1 and re-Hst1 had the same apparent molecular weights, while cationic PAGE showed that re-Hst1 was more basic. Phosphate analysis indicated 1 mol phosphate/mol of native histatin 1, while re-Hst1 lacked any detectable phosphate. Re-Hst1 demonstrated candidacidal activity comparable to that of native histatin 1, but displayed substantially lower binding to hydroxyapatite. These results show that phosphorylation of histatin 1 at residue 2 contributes significantly to its ability to bind to hydroxyapatite.

Antifungal Agents↗
PubMed1995

Identifying tumor suppressors in genetic mosaics: the Drosophila lats gene encodes a putative protein kinase.

We have identified recessive overproliferation mutations by screening and examining clones of mutant cells in genetic mosaics of the fruitfly Drosophila melanogaster. This type of screen provides a powerful approach for identifying and studying potential tumor suppressors. One of the identified genes, lats, has been cloned and encodes a putative protein kinase that shares high levels of sequence similarity with three proteins in budding yeast and Neurospora that are involved in regulation of the cell cycle and growth. Mutations in lats cause dramatic overproliferation phenotypes and various developmental defects in both mosaic animals and homozygous mutants.

Amino Acid Sequence↗
PubMed1995

[Lymphatic interventional radiology in the treatment of lymphatic neoplasmas].

We treated 22 cases of lymphatic neoplasma with chemotherapy via lymphatic ducts under the control of X-ray monitor, a method we called interventional trans-lymphatic chemotherapy. The method includes: (1) lymphography, (2) trans-lymphatic infusion with anticancer agents, (3) lavage of lymphatic vessel, (4) treatment of complications. Six cases of primary lymphoma and 16 cases of lymph-metastatic carcinoma were treated this way. The short-term results were excellent. Main symptoms and signs disappeared in 19 cases (19/22), and markedly improved in 2. X-ray features of 18 cases showed that enlarged lymph nodes returned to normal after the therapy. Follow-up for 28 months showed no recurrence.

Adult↗
PubMed1995

[The changing of orofacial structure by bite opening with Begg technique].

Cephalometric study of 30 deep-bite cases treated with extraction of four first premolars and Begg stage I appliance shows that the bite opening consists of not only intrusion of the upper and lower incisors but also extrusion of the upper and lower molars, furthermore, the lower incisors depressed more than the upper ones and the lower molars are elevated more than the upper ones. The moving type of teeth, the changing of base bone and soft tissue profile are also discussed in this article.

Adolescent↗
PubMed1995

Cholecystokinin octapeptide reverses mu-opioid-receptor-mediated inhibition of calcium current in rat dorsal root ganglion neurons.

Cholecystokinin octapeptide (CCK-8) is reported to antagonize the analgesic effect produced by mu- and kappa- but not delta-opioid agonist in spinal cord. However, the mechanisms of interaction remain obscure. In the present study, whole-cell patch-clamp recording was performed on acutely isolated rat dorsal root ganglion (DRG) neurons to evaluate the effects of the highly specific mu-opioid agonist ohmefentanyl and the delta-opioid agonist DPDPE on voltage-gated calcium channels and the possible interaction between CCK-8 receptor and mu- or delta-opioid receptor. The results indicated that ohmefentanyl, but not DPDPE, can suppress the voltage-gated calcium currents elicited in DRG neurons, an effect readily reversed by naloxone or by the antiopioid peptide CCK-8. The effect of CCK-8 can in turn be abolished by the CCK-B receptor antagonist L365,260. CCK-8 used by itself has no enhancing effect, but rather a depressant effect, on calcium currents. However, used simultaneously with ohmefentanyl, CCK-8 shows a clear-cut reversal of depression of the mu-opioid. We conclude that the depressant effect produced by mu-opioid on voltage-gated calcium current in DRG neurons can be antagonized by CCK-8 through CCK-B receptor located in the same neuron. The delta-opioid DPDPE has no direct effect on the voltage-gated calcium current in DRG neurons.

Animals↗
PubMed1995

[Influence of changing Ca++ concentration in neiguan (PC 6) on the effect of acupuncture treating experimental arrhythmia of rabbits].

We have made two kinds of experimental arrhythmia of rabbits by injecting aconitine and stimulating hypothalamus. Acupuncture could improve arrhythmia, after Ca++ in Neiguan (PC6) was chelated with EDTA solution the effect of acupuncturing Neiguan (PC6) was abolished. It indicates that Ca++ may be the key factor of acupuncture effect and one of the important material bases of the functional activity of meridians and collaterals.

Aconitine↗
PubMed1994

A member of the Notch group of interacting loci, deltex encodes a cytoplasmic basic protein.

Prior genetic studies have suggested a functional relationship between the product of the deltex gene and those of three of the so-called "neurogenic" loci, Notch, Delta and mastermind. To gain further insight into this relationship, we have proceeded with a molecular characterization of deltex. We report that deltex encodes a maternally and zygotically expressed transcript that conceptually translates to a basic protein of novel sequence. Immunolocalization of the protein reveals an apparently ubiquitous distribution in embryonic and imaginal tissues. Because our detection methods also reveal a very low level of protein accumulation within the cytoplasm of cells, we have used transgenic flies to confirm this observation by ectopically expressing deltex under the control of a heat shock gene promoter. The resulting overexpression rescues deltex mutant defects but does not produce any obvious phenotypic abnormalities in otherwise wild-type flies. Finally, we examine genetically several Supressor of deltex mutations for evidence of functional integration with deltex and other neurogenic genes. We demonstrate that in addition to suppressing all adult morphological defects of deltex alleles, these suppressors also are capable of suppressing most synergistic effects involving deltex and Notch, Delta and mastermind.

Amino Acid Sequence↗