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Biomedical subjects

H Jiang

Publications and source records attributed to H Jiang.

At least 397 records · Page 22Linked to original sources

The gene 32 single-stranded DNA-binding protein is not bound stably to the phage T4 presynaptic filament.

A central reaction in homologous recombination is synapsis, which involves invasion of duplex DNA by a homologous single strand. A key intermediate in this process is the presynaptic filament, a protein-DNA complex composed of a "strand transferase" polymerized along the invading single strand. In this report, the organization and mechanism of assembly of the bacteriophage T4 presynaptic filament are explored. Three T4 proteins, encoded by the uvsX, uvsY and 32 genes, are involved in this process. It is demonstrated that a well-defined series of events involving multiple protein-DNA and protein-protein interactions is required to mediate a transition from an initial gene 32-DNA complex to a mature presynaptic filament in which the UvsX and UvsY proteins are in contact with the DNA and each other, while most or all of the gene 32 protein is removed from the complex.

Bacteriophage T4↗

Suppression of human ribosomal protein L23A expression during cell growth inhibition by interferon-beta.

Interferons inhibit cell growth in normal and tumor-derived cells. The molecular basis of interferons antiproliferative activity remains to be defined. Using subtraction hybridization, a human melanoma differentiation associated gene, mda-20, has been identified that is down-regulated by treatment with interferon. Sequence analysis indicates that mda-20 is human ribosomal protein L23a (rp L23a). The mRNA levels of rp L23a and growth are diminished in a variety of human tumor cell lines following treatment with human fibroblast interferon, interferon-beta (IFN-beta). Expression of rp L23a is also reduced in human melanoma cells treated with human leukocyte (IFN-alpha) and immune (IFN-gamma) interferons, but not by growth inhibition resulting from serum starvation. These findings suggest that growth suppression alone is not sufficient to reduce rp L23a expression. Instead, reduced rp L23a mRNA results from biochemical changes mediated by interferons. Ectopic expression of an antisense rp L23a sequence in human HeLa cervical carcinoma cells results in a reduction in colony formation indicating a direct antiproliferative effect by inhibiting rp L23a expression. The mechanism underlying inhibition in rp L23a expression in IFN-beta-treated cells may involve antisense rp L23a RNA. These results suggest that rp L23a may be one of the target molecules involved in mediating growth inhibition by interferon.

Adenocarcinoma↗

Characterization and functional analysis of 12 naturally occurring reactive site variants of serpin-1 from Manduca sexta.

Serpin gene-1 from the tobacco hornworm, Manduca sexta, encodes, through alternative exon usage, 12 reactive site variants (Jiang, H., Wang, Y. and Kanost, M. R., (1994) J. Biol. Chem. 269, 55-58; Jiang, H., Wang, Y., Huang, Y., Mulnix, A. B., Kadel, J., Cole, K., and Kanost, M. R. (1996) J. Biol. Chem. 271, 28017-28023). These 43-kDa proteins differ from each other only in their COOH-terminal 39-46 residues, which include the reactive site. To test the hypothesis that these proteins are proteinase inhibitors of diverse selectivities and to begin to elucidate their physiological functions, we expressed the 12 serpin-1 variants in Escherichia coli. Seven of the variants inhibited mammalian serine proteinases, with association rate constants comparable with those of human serpins. Serpin-1A, with a P1 Arg residue, inhibited both trypsin and plasmin. Serpin-1B (P1 Ala) and serpin-1F (P1 Val) inhibited porcine pancreatic elastase and human neutrophil elastase. Serpin-1H, -1K, and -1Z, all with a Tyr residue at the P1 position, inhibited chymotrypsin and cathepsin G. Serpin-1I (P1 Leu) inhibited both elastase and chymotrypsin. Nine of the serpin variants were active as inhibitors of microbial serine proteinases, including subtilisin Carlsberg, proteinase K, and two proteinases secreted by an entomopathogenic fungus, Metarhizium anisopliae. In addition, one of the serpin variants, serpin-1J, strongly inhibited activation of M. sexta hemolymph phenoloxidase, a pathway involving a serine proteinase cascade. This pathway is a component of the defensive response of insects to microbial infection. These results suggest that the products of M. sexta serpin gene-1 may be important in regulating both exogenous and endogenous serine proteinases in hemolymph.

Animals↗

Identification of the location, extent, and pathway of sensory neurologic feedback after mechanical stimulation of a lateral spinal ligament in chickens.

STUDY DESIGN: This study traced the location, extent, and pathway of sensory feedback after the mechanical stretching of a lateral spinal ligament in young chickens. The pathway was traced by locating the sites of Fos protein production in neuronal cell bodies at various sites in the nervous system. OBJECTIVES: To trace the location, extent, and pathway of sensory feedback after the mechanical stretching of a lateral spinal ligament in young chickens. SUMMARY OF BACKGROUND DATA: The innervation of ligaments is thought to form part of a protective feedback mechanism to provide stability for joints. The precise pathway and extent of the feedback for spinal ligaments is currently unknown. Such information would provide a clear focus for future studies, especially for diseases such as scoliosis where it has been suggested that there is abnormality in perception of sensory feedback. METHODS: The intertransverse ligament on the right side at T3-T4 in 4-week-old chickens was exposed by blunt dissection. After Fos production resulting from the surgery had been stopped, the ligament was stretched mechanically and repeatedly for 60 minutes using a 300-g weight. Various areas of the nervous system then were sectioned and processed immunohistochemically to identify areas of Fos production in nerve cell bodies. The presence of Fos indicated neurons that had been stimulated by the stretching the ligament, including interneurons along the feedback pathway. RESULTS: Fos protein was identified in nerve cell bodies in the dorsal root ganglia and intermediate gray matter of the spinal cord at the level of stimulation as well as at several spinal cord levels above and below the site of stimulation. Identification was made on the ipsilateral and the contralateral sides, although the extent of Fos production was less on the contralateral side. Fos presence also was identified in sympathetic ganglia at these sites. Nerve cell bodies in the combined nucleus cuneatus and gracilis in the medulla oblongata, the vestibular nuclei, and the thalamus also contained Fos-positive particles. CONCLUSIONS: Stretching a single lateral ligament of the spine produces a barrage of sensory feedback from several spinal cord levels on both sides of the spinal cord. This sensory information also is transferred to higher levels in the brain, including the nucleus gracilis and cuneatus, the vestibular nuclei, and the thalamus. These sites of Fos production suggest the locations of pathways for this sensory information, which include the dorsal columns and the spinocerebellar tracts. The information obtained from this study provides a clear focus for future studies in this area, particularly for diseases such as scoliosis where it is thought that incorrect perception of sensory information from the ligaments might be a major contributing factor.

Animals↗

A multiple echo pulse sequence for diffusion tensor imaging and its application in excised rat spinal cords.

A new imaging sequence for rapid determination of the apparent self-diffusion tensor of water was developed and tested on fixed excised rat spinal cords. To reduce the time required to determine the tensor, the sequence utilized a new single-shot approach with multiple spin echoes. An assumption of cylindrical symmetry in the sample was made, thus requiring the measurement of only four of the six unique elements of the tensor. This assumption was found experimentally to be valid, and the results obtained using the new sequence were found to be quantitatively the same as results obtained using a standard spin-echo sequence.

Animals↗

Significance of proliferating cell nuclear antigen expression in liver metastasis of colorectal cancer.

PURPOSE: The study contained herein was aimed at finding some possible pathologic factors that have significance for the prediction of liver metastasis in colorectal cancer. METHOD: Resected specimens of colorectal cancer from 23 patients with liver metastasis and 30 patients without liver metastasis were subjected to pathologic study, including microscopic characteristics and proliferating cell nuclear antigen immunohistochemistry assay. RESULTS: Strongly positive expression of proliferating cell nuclear antigen was present in 65.21 percent (15/23) of the liver metastasis group, whereas it was found in only 20 percent (6/30) of the group without liver metastasis (P < 0.005). Deeper invasion to the muscularis propria or serosa and less infiltration of lymphocytes surrounding the tumor were more frequently found in the liver metastasis group than in the other group (P < 0.025). CONCLUSION: Extent of proliferating cell nuclear antigen expression, depth of invasion, and reaction of lymphocyte infiltration of the primary tumor could have predictive significance of colorectal cancer in liver metastasis.

Adult↗

Sequence and expression of mouse proteasome activator PA28 and the related autoantigen Ki.

Using reverse transcription polymerase chain reaction, we cloned and sequenced full-length mouse cDNAs for the two homologous subunits of the proteasome activator PA28 (PA28alpha and PA28beta), as well as for the related protein Ki. These proteins are highly conserved among species. Northern blot analysis of PA28a, PA28b, and Ki mRNA demonstrated broad tissue distribution. Although single transcripts were detected for PA28a and PA28b, two different sized transcripts were detected for mouse Ki, suggesting either alternative splicing or alternate polyadenylation sites. The levels of these transcripts increased in response to interferon-gamma (IFN-gamma) treatment in mouse H6 hepatoma cells, although PA28a and PA28b were induced to a greater extent than Ki, and the effect of IFN-gamma stimulation on Ki expression was transient. Southern blot analysis suggests that both PA28a and PA28b are multiple-copy genes, while Ki is a single-copy gene.

Amino Acid Sequence↗

Transient effects of MK-801 administration on secretion of luteinizing hormone and prolactin in ovariectomized and estradiol-treated sheep.

The neuroendocrine role of endogenous ligands for the excitatory amino acid receptor subtype known as the NMDA receptor was investigated by administering the NMDA receptor antagonist MK-801 to ovariectomized (ovx) and estradiol-treated sheep. Repetitive administration of MK-801 at intravenous (iv) dosages of 0.1 mg/kg to untreated ovx ewes did not affect the episodic profiles of luteinizing hormone (LH) release, but each injection of MK-801 abruptly stimulated release of prolactin (PRL) demonstrating the effectiveness of the dosage. Injection of estradiol-17beta (50 microg/ewe) to ovx ewes produced the expected biphasic LH response; an initial suppression followed by a surge-like LH increase together with an elevated basal secretion of PRL. Injections of MK-801 occurring 9 and 11 h after estradiol-17beta injection rapidly and transiently increased serum LH in a very unexpected response. However, these same MK-801 injections also resulted in decreased serum LH 14-17 h after estradiol-17beta by delaying the onset of the surge-like release of LH. Estradiol-17beta administration itself elevated basal release of PRL to serum concentrations observed after repetitive MK-801, and further injection of MK-801 no longer transiently increased serum PRL as it had done prior to estradiol-17beta treatment. In summary, antagonizing the endogenous excitatory amino acid ligands of the NMDA receptor with MK-801 did not alter either the timing or magnitude of the putative episodic discharges of gonadotropin-releasing hormone (GnRH) which in turn cause the episodic releases of pituitary LH in ovx sheep. Estrogen administration created a transient neuroendocrine environment in which antagonism of these endogenous ligands was stimulatory to abrupt discharge of GnRH and thereby to acute release of LH. Antagonism of NMDA receptors in untreated ovx ewes stimulated release of PRL suggesting that the endogenous NMDA ligands were probably stimulatory to the release of a PRL-inhibiting neurohormone such as dopamine.

Animals↗

Distinct tissue and cellular distribution of two major isoforms of calcineurin.

The protein phosphatase calcineurin is known to be an essential intracellular signal transducer involved in the TCR-mediated signal transduction pathway and is the common target of the immunosuppressive drugs cyclosporin A (CsA) and FK506. The catalytic subunit of calcineurin exists in multiple isoforms, but their functional differences are not known. It has been assumed that the alpha isoform of calcineurin is the relevant isoform mediating TCR signaling. Recently, calcineurin alpha was knocked out in mice, but no defect in the TCR-mediated IL-2 production was observed, suggesting that another isoform of calcineurin mediates the TCR signal transduction pathway. We have generated specific polyclonal antibodies against the alpha and the beta2 isoforms of calcineurin and examined their distribution in murine tissues and immune cells by immunohistochemical staining and Western blot analysis. We found that the beta2 isoform of calcineurin is predominant in T and B lymphocytes as well as in thymus compared to the alpha isoform, suggesting that the beta2 isoform may play a key role in TCR signaling. Furthermore, we observed that the two isoforms exhibit distinct expression patterns in both kidney and thymus, indicating that the two isoforms of calcineurin have distinct cellular functions. Together, these findings raise the possibility that the nephrotoxicity associated with CsA and FK506 can be reduced by designing novel inhibitors of calcineurin that target specific isoforms of the enzyme.

Amino Acid Sequence↗

Positional, directional and speed selectivities in the primary auditory cortex of the cat.

Responses of high-frequency primary auditory cortex (A1) neurons of the cat to noise stimulation were obtained in a quasianechoic chamber using a static and an apparently moving stimulus presented at similar azimuths. Simulated motion toward right or left as well as different simulated velocities were used. Under static stimulation, most units were contralateral-preferring followed by ipsilateral- and midline-preferring. Some were omnidirectional and a few were unclassifiable. Width of tuning was similar for contralateral-, ipsilateral- and midline-preferring units. Overall, about 25% were finely tuned (< 20 degrees) and the remaining were broadly tuned (> or =20 degrees). All cells sampled with static stimulation responded to apparent motion. About one quarter of the units were sensitive to the direction of the simulated moving noise in that they responded at least twice as much to one direction as to the other. Almost all directional contralateral-preferring units responded more when the apparent motion was directed toward ipsilateral azimuths, whereas all directional ipsilateral-preferring units responded preferentially to contralaterally oriented motion. In some units, up to five apparent speeds were tested. About half the units were not speed-selective (46%). The other cells were tuned to a preferential speed (40%), decreased their response as the apparent speed increased (10%) or displayed direction-dependent speed selectivity (4%). These results indicate that moving-sound sources are processed by some A1 single units.

Acoustic Stimulation↗

A software system for interactive MR signal processing.

This paper describes a new software platform called "V" which is designed for interactive processing of magnetic resonance data. The package currently contains a variety of functions for image reconstruction and spectral data analysis. More importantly, it provides a framework to facilitate the exchange and application of processing algorithms among researchers in the field. We discuss the architecture of the package, give an example to demonstrate its operation and provide instructions for obtaining the software.

Algorithms↗

Molecular cloning of cDNAs for two pro-phenol oxidase subunits from the malaria vector, Anopheles gambiae.

Phenol oxidase exists in insect hemolymph as a zymogen, pro-phenol oxidase (pro-PO), which is activated by specific proteolysis in response to infection or wounding. Phenol oxidase catalyses the synthesis of quinones that polymerize to form melanin deposits, which encapsulate parasites and help to seal wounds. Antibodies to pro-PO from Manduca sexta bound to 76, 72, and 71 kDa polypeptide bands from hemolymph of Anopheles gambiae larvae. This antiserum was used to screen a cDNA library from A. gambiae fourth-instar larvae. Full-length clones were isolated for two different pro-POs, designated A. gambiae proPO-p1 and proPO-p2, which are 67% identical in nucleotide sequence and 66% identical in deduced amino acid sequence. The A. gambiae pro-PO sequences are more similar to pro-PO from Drosophila melanogaster than to lepidopteran or crustacean pro-PO sequences in the GenBank database. Like the other arthropod pro-POs, the A. gambiae pro-PO sequences lack a signal peptide and have two conserved regions predicted to bind two copper atoms in the active site of the enzyme. The availability of these pro-PO cDNAs should be useful in examining the biochemical differences between A. gambiae strains that are refractory or susceptible to Plasmodium infection, and differ in their ability to encapsulate the parasites.

Amino Acid Sequence↗

Subunit composition of pro-phenol oxidase from Manduca sexta: molecular cloning of subunit ProPO-P1.

Phenol oxidase (PO) is known to play an important role in defense mechanisms in insect immunity. It is present as a zymogen in insect hemolymph, and can be activated by a specific proteolytic reaction that is stimulated by microbial cell wall components. The pro-phenol oxidase (pro-PO) purified from the larval hemolymph of Manduca sexta contains two polypeptides in equal amounts as revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A cDNA for one of the polypeptides, now designated proPO-p2, has been isolated (Hall et al. (1995) Proc. Natl. Acad. Sci. USA, 92, 7764-7768). We purified pro-PO from plasma of M. sexta and characterized its subunit composition. A cDNA for M. sexta proPO-p1 was isolated from a larval hemocyte cDNA library. M. sexta proPO-p1 is 78% identical in amino acid sequence to Bombyx mori proPO-p1, but only 50% to M. sexta or B. mori proPO-p2. Immunofluorescence labelling and in situ hybridization showed that the pro-PO is synthesized in a single hemocyte type, the oenocytoids. Analysis of pro-PO by size exclusion high-pressure liquid chromatography (HPLC) revealed that pro-PO exists as monomeric, dimeric, trimeric or multimeric structures depending on the ionic strength. All of these isoforms of the protein have phenol oxidase activity upon activation with a detergent, cetylpyridinium chloride. In analysis by non-denaturing PAGE, the majority of the purified pro-PO was present as two dimers of distinct mobility (fast and slow forms). Both forms contain proPO-p1 and proPO-p2, suggesting that they are heterodimers. Individual larvae can contain the slow form, the fast form, or both, which suggests that the slow and fast forms of proPO are allelic variants. These results indicate that there are two pro-PO genes in M. sexta, which are coordinately expressed in oenocytoids, and whose products form predominantly heterodimers in plasma.

Amino Acid Sequence↗

Structure-binding relation of philanthotoxins from nicotinic acetylcholine receptor binding assay.

Philanthotoxins are noncompetitive inhibitors of the nicotinic acetylcholine receptor and the various glutamate receptors. Analogues carrying photoaffinity labels, fluorine atoms for solid-state NMR studies of ligand/receptor interaction, and large head groups such as porphyrins and planar bulky aromatic rings (BIG analogues) for clarifying mode of entry and orientation of analogues in receptors have been synthesized, assayed against the nicotinic acetylcholine receptor, and brief comments are given for the assay results.

Amino Acids↗

Effects of antisense p21 (WAF1/CIP1/MDA6) expression on the induction of differentiation and drug-mediated apoptosis in human myeloid leukemia cells (HL-60).

The p21MDA6 gene product induces cell cycle arrest in p53-null human leukemic cells exposed to differentiation stimuli. We employed an HL-60 cell line stably transfected with a p21MDA6 antisense construct to compare the effects of p21MDA6 dysregulation on the response of myeloid leukemia cells to differentiating and cytotoxic agents. Antisense-expressing cells (HL-60/AS5) treated with 5 nM PMA for 24 h exhibited attenuated induction of p21MDA6 compared to empty vector controls (HL-60/V2). This phenomenon was accompanied by a reduction in the percentage of cells undergoing G1 arrest (67.6 +/- 4.7 vs 82.9 +/- 1.3; P < or = 0.01) and expressing the monocytic maturation marker cd11b (35.5 +/- 2.8 vs 50.5 +/- 2.4; P < or = 0.005). Although HL-AS5 and HL-60/V2 cells did not exhibit obvious differences in the phosphorylation status of the retinoblastoma protein (pRB), in E2F complex formation, or in p27klp1 induction following PMA exposure, inhibition of activity of cyclin-dependent kinase-2 was attenuated in the antisense-expressing line. A 24-h exposure to 5 nM PMA also reduced the cloning efficiency of HL-60/V2 cells to a significantly greater extent than HL-60/AS5 cells (ie to 30.1 +/- 7.0 vs 57.2 +/- 5.6 of controls; P < or = 0.01). In contrast to the disparate responses to PMA, HL-60/AS5 and HL-60/V2 cells treated with the antimetabolite 1-beta-D-arabinofurano-sylcytosine (Ara-C; 10 microM for 6 h) displayed equal susceptibility to G1 arrest, apoptosis, and inhibition of clonogenicity, phenomena unaccompanied by p21MDA6 and p27klp1 induction, or pRB dephosphorylation. These observations indicate that dysregulation of p21MDA6 in p53-null human myeloid leukemia cells interferes with PMA-related G1 arrest, CDK-2 inhibition, differentiation, and loss of clonogenic survival in the absence of obvious alterations in pRB phosphorylation status or E2F complex formation. They also provide functional evidence that p21MDA6 induction does not appear to be required for Ara-C-induced apoptosis, G1 arrest, or the resulting reduction in the self-renewal capacity of HL-60 cells.

Apoptosis↗

Case-control assessment of diet and lung cancer risk in African Americans and Mexican Americans.

In this case-control study we determined whether dietary differences underlie some of the ethnic and sex differences in US lung cancer rates. We examined the relationship between diet and lung cancer development in 137 lung cancer cases (93 African Americans and 44 Mexican Americans) and 187 controls (78 African Americans and 109 Mexican Americans). Cases reported a higher daily mean total fat intake (p < 0.001), whereas controls had a higher daily mean intake of dietary fiber (p < 0.001) and fruits (p = 0.02). Ethnic differences in diet were also observed: Mexican Americans consumed less total fat (p < 0.02) and more fiber (p < 0.001) and vegetables (p = 0.08) than African Americans. Additionally, men consumed more total fat (p = 0.08) and less fiber (p = 0.001), fruits (p < 0.001), and vegetables (p = 0.002) than women. Multivariable analysis, after adjustment for the effects of pack-years of smoking, age, total energy intake, sex, and ethnicity, demonstrated a positive association between high total fat consumption and lung cancer risk (p < 0.01) and an inverse association between high fruit consumption and lung cancer risk (p = 0.05). In conclusion, our findings support the hypothesis that diet, particularly high fat consumption and low fruit and vegetable consumption, contributes (independent of cigarette smoking) to the excess lung cancer risk in African-American men, who have the highest lung cancer rates in the United States.

Black or African American↗

Buspirone treatment of psychiatrically hospitalized prepubertal children with symptoms of anxiety and moderately severe aggression.

Open-label buspirone was studied in 25 prepubertal psychiatric inpatients (age 8.0 +/- 1.8 years, 76% boys) presenting with anxiety symptoms and moderately aggressive behavior. Patients with severe aggression, requiring rapid treatment with mood stabilizers or neuroleptics, were excluded. A 3-week titration (maximum 50 mg daily) preceded a 6-week maintenance phase at optimal dose. Buspirone was discontinued in 6 children (25%): 4 developed increased aggression and agitation, and 2 developed euphoric mania. For the 19 patients who completed the study, mean optimal dose was 28 mg daily. Among completers, depressive symptoms were reduced 52% by Week 6 on Children's Depression Inventory (p < or = 0.001). Decreased aggressivity was reflected in a 29% reduction on Measure of Aggression, Violence, and Rage in Children [MAVRIC] ratings (p < or = 0.02) and in 86% less time in seclusion or physical restraints (p < or = 0.02). Clinical Global Assessment scores improved (CGAS 41 vs. 54, p < or = 0.01). Only 3 children improved sufficiently to continue buspirone after the study. Residual aggressivity and global functioning remained problematic. Buspirone may pose behavioral risks in treating moderate aggressivity in 24% of children with anxiety; in the others, the therapeutic effects on aggression, anxiety, and depression were limited but significant.

Aggression↗