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Biomedical subjects

Z Li

Publications and source records attributed to Z Li.

At least 901 records · Page 50Linked to original sources

KRC transcripts: identification of an unusual alternative splicing event.

Mouse KRC is a large zinc finger protein that binds to the kappaB motif of gene transcription and to the recognition signal sequences for the somatic recombination of the immunoglobulin and T-cell receptor gene segments. The mouse KRC gene is more than 70 kilobases (kb) in size, and contains at least seven exons, with the largest transcript being approximately 9.5 kb. Multiple differentially spliced transcripts of KRC were identified in thymus and brain, which would result in the production of multiple KRC protein isoforms with different N-termini and number of DNA binding domains. Alternative splicing events leading to the production of these multiple transcripts have been elucidated. Of particular interest are the exclusions in some transcripts of sequences from a gigantic exon of 5487 base pairs (bp), or from an exon of 176 bp. Both potentially deleted exons code for zinc finger motifs that are essential components of the N-terminal and C-terminal DNA binding domains, respectively. Another intriguing phenomenon found in some KRC transcripts is the skipping of a 459 bp fragment within the gigantic exon that would code for the N-terminal DNA binding domain. Bacterial fusion proteins derived from this fragment bind specifically to KRC target DNAs. Apparently, distinct alternative splicing events could eliminate the N-terminal DNA binding domain of KRC.

Alternative Splicing↗

Glucose repression on RIM1, a gene encoding a mitochondrial single-stranded DNA-binding protein, in Saccharomyces cerevisiae: a possible regulation at pre-mRNA splicing.

The mitochondrial single-stranded DNA-binding protein (SSB) encoded by anuclear gene, RIM1, is a homolog of Escherichia coli SSB. The addition of glucose decreased the amount of RIM1-mRNA in cells growing in a glycerol medium, but increased the amount of the immature RIM1-mRNA. The changes in the amounts of both mature and immature RIM1-mRNAs were dependent on SRN1/REG1/HEX2, a gene relating to pre-mRNA-splicing and glucose repression. These observations suggest that the expression of the mitochondrial SSB is regulated, at least in part, by pre-mRNA splicing under the control of glucose repression.

Culture Media↗

Localization of the Drosophila checkpoint control protein Bub3 to the kinetochore requires Bub1 but not Zw10 or Rod.

We report here the isolation and molecular characterization of the Drosophila homolog of the mitotic checkpoint control protein Bub3. The Drosophila Bub3 protein is associated with the centromere/kinetochore of chromosomes in larval neuroblasts whose spindle assembly checkpoints have been activated by incubation with the microtubule-depolymerizing agent colchicine. Drosophila Bub3 is also found at the kinetochore regions in mitotic larval neuroblasts and in meiotic primary and secondary spermatocytes, with the strong signal seen during prophase and prometaphase becoming increasingly weaker after the chromosomes have aligned at the metaphase plate. We further show that the localization of Bub3 to the kinetochore is disrupted by mutations in the gene encoding the Drosophila homolog of the spindle assembly checkpoint protein Bub1. Combined with recent findings showing that the kinetochore localization of Bub1 conversely depends upon Bub3, these results support the hypothesis that the spindle assembly checkpoint proteins exist as a multiprotein complex recruited as a unit to the kinetochore. In contrast, we demonstrate that the kinetochore constituents Zw10 and Rod are not needed for the binding of Bub3 to the kinetochore. This suggests that the kinetochore is assembled in at least two relatively independent pathways.

Amino Acid Sequence↗

The Des-1 protein, required for central spindle assembly and cytokinesis, is associated with mitochondria along the meiotic spindle apparatus and with the contractile ring during male meiosis in Drosophila melanogaster.

Spermatogenesis in Drosophila melanogaster serves as an excellent model system for the isolation and analysis of genes required in the control of chromosome segregation and cytokinesis. We report here the isolation and molecular characterization of a novel P-element induced allele of the des-1 gene, which leads to male sterility as a consequence of the failure of central spindle assembly in meiotic spermatocytes and the formation of aberrant meiotic end products characteristic of cytokinesis failure. We have raised affinity-purified antibodies against a Des-1 fusion protein, and localized the Des-1 protein in Drosophila spermatocytes. We show that the Des- protein is colocalized with mitochondria throughout male meiosis, becoming intimately associated with mitochondria along the spindle apparatus during anaphase and telophase, and with the Nebenkern, or mitochondrial derivative, of the meiotic end products. In addition, a significant association of Des-1 with the contractile ring is observed during anaphase and telophase of meiosis. These observations, together with the presence of six potential transmembrane domains in the Des-1 protein, raise the possibility that Des-1 may act as part of an anchoring mechanism that links membrane-bounded cellular compartments to components of the cytoskeleton.

Animals↗

Complete physical map of the common deletion region in Williams syndrome and identification and characterization of three novel genes.

Williams syndrome (WS) is a contiguous gene deletion disorder caused by haploinsufficiency of genes at 7q11.23. We have shown that hemizygosity of elastin is responsible for one feature of WS, supravalvular aortic stenosis (SVAS). We have also implicated LIM-kinase 1 hemizygosity as a contributing factor to impaired visual-spatial constructive cognition in WS. However, the common WS deletion region has not been completely characterized, and genes for additional features of WS, including mental retardation, infantile hypercalcemia, and unique personality profile, are yet to be discovered. Here, we present a physical map encompassing 1.5 Mb DNA that is commonly deleted in individuals with WS. Fluorescence in situ hybridization analysis of 200 WS individuals shows that WS individuals have the consistent deletion interval. In addition, we identify three novel genes from the common deletion region: WS-betaTRP, WS-bHLH, and BCL7B. WS-betaTRP has four putative beta-transducin (WD40) repeats, and WS-bHLH is a novel basic helix-loop-helix leucine zipper (bHLHZip) gene. BCL7B belongs to a novel family of highly conserved genes. We describe the expression profile and genomic structure for each of these genes. Hemizygous deletion of one or more of these genes may contribute to developmental defects in WS.

Amino Acid Sequence↗

Rice ragged stunt oryzavirus genome segment S4 could encode an RNA dependent RNA polymerase and a second protein of unknown function.

The complete nucleotide sequence of genome segment S4 of rice ragged stunt oryzavirus (RRSV, Thai-isolate) was determined. The 3823 bp sequence contains two large open reading frames (ORFs). ORF1, spanning nucleotides 12 to 3776, is capable of encoding a protein of M(r) 141,380 (P4a). The P4a amino acid sequence predicted from the nucleotide sequence contains sequence motifs conserved in RNA-dependent RNA polymerases (RDRPs). When compared for evolutionary relationships with RDRPs of other reoviruses using the amino acid sequences around the conserved GDD motif, P4a was shown to be more related to Nilaparvata lugens reovirus and reovirus serotype 3 than to rice dwarf phytoreovirus, bovine rotavirus or bluetongue virus. The ORF2, spanning nucleotides 491 to 1468, is out of frame with ORF1 and is capable of encoding a protein of 36,920 (P4b). Coupled in vitro transcription-translation from cloned ORF2 in wheat germ extract confirmed the existence of ORF2 but in vivo production and possible function of P4b is yet to be determined.

Amino Acid Sequence↗

Anti-Ro/SS-A positivity and heat shock protein antibodies in patients with normal-pressure glaucoma.

PURPOSE: To describe the clinical and laboratory findings in 10 patients with normal-pressure glaucoma and anti-Ro/SS-A positivity by enzyme-linked immunosorbent assay (ELISA) and to determine whether that positivity may be related to an autoimmune mechanism for the optic neuropathy. METHODS: In this prospective study, we evaluated ocular and systemic clinical findings of 10 patients with normal-pressure glaucoma and anti-Ro/SS-A positivity by ELISA, including sicca complex features. Oüchterlony immunodiffusion was performed to confirm the presence of antibodies for Ro/SS-A, and the presence of other serum antibodies and their possible cross-reactivities with Ro/SS-A were investigated. RESULTS: None of the 10 patients with normal-pressure glaucoma and anti-Ro/SS-A positivity (by ELISA) had clinical or laboratory signs of Sjögren syndrome or other connective tissue diseases. Only one of 10 patients had evidence of anti-Ro/SS-A antibodies by Oüchterlony immunodiffusion. All patients demonstrated serum immunoreactivity to bacterial heat shock protein 60 (hsp60) by Western blotting. Cross-reactivity between bacterial hsp60 and Ro/SS-A was demonstrated by Western blotting. Immunoreactivity to bacterial hsp60 by ELISA was significantly elevated in the sera of patients with normal-pressure glaucoma. Furthermore, patients with either normal-pressure or primary open-angle glaucoma had increased serum immunoreactivity to human hsp60. CONCLUSIONS: Anti-Ro/SS-A positivity by ELISA in 10 patients with normal-pressure glaucoma was associated with a high level of serum immunoreactivity to bacterial hsp60, which may indicate that their glaucomatous optic neuropathy involves an as yet unidentified autoimmune mechanism. The identification of autoantibodies that react with human hsp60 in patients with normal-pressure and primary open-angle glaucoma may signify a common finding associated with the glaucomatous optic neuropathy process in some patients and appears to be unrelated to intraocular pressure levels.

Adult↗

Blood flows and metabolic components of the cardiome.

This is a plan for the first stage of The Cardiome Project. The cardiome is the representation, in quantitative, testable form, of the functioning of the normal heart and its responses to intervention. The goal is to integrate the efforts of many years into a comprehensive understandable scheme. Past efforts have spanned the fields of transport within blood vessels, the distributions of regional coronary blood flows, permeation processes through capillary and cell walls, mediated cell membrane transport, extra- and intracellular diffusion, cardiac electrophysiology, the uptake and metabolism of the prime substrates (fatty acid and glucose), the metabolism of the purine nucleosides and nucleotides (mainly adenosine and ATP), the regulation of the ionic currents and of excitation-contraction coupling and finally the regulation of contraction. The central theme is to define the coronary flows and metabolic components of a computer model that will become a part of a three-dimensional heart with appropriate fibre shortening and volume ejection. The components are: (a) coronary flow distributions with appropriate heterogeneity, (b) metabolism of the substrates for energy production, (c) ATP, PCr and energy metabolism and (d) calcium metabolism as it relates to excitation-contraction coupling. The modeling should provide: (1) appropriate responses to regional ischemia induced by constriction of a coronary artery, including tissue contractility loss and aneurysmal dilation of the ischemic region; (2) physiological responses to rate changes such as treppe and changes in metabolic demand and (3) changes in local metabolic needs secondary to changes in the site of pacing stimulation and shortening inactivation or stretch activation of contraction.

Adenosine↗

Modification of pentasaccharide core of surface N-glycans during differentiation of HL-60 cells.

All-trans retinoic acid (ATRA) or phorbol-12-myristate-13-acetate (PMA) are two representative inducers which induce the differentiation of HL-60 cells to myelocytes and monocytes respectively. Structural modifications of the pentasaccharide core portion of N-glycans on cell surface glycoproteins during differentiation of HL-60 cells induced by these two agents and the enzymatic mechanism of the structural modifications were studied. Both ATRA and PMA enhanced the incorporation of [3H]-labeled mannose into N-glycans of glycoproteins from cell surface. By using E-PHA and WGA lectin affinity chromatography to analyze [3H]-N-glycans prepared from the surface of control, ATRA- and PMA-treated cells, it was found that the content of bisecting GlcNAc (bis-GlcNAc) attached to the core of N-glycans decreased in ATRA-treated cells, but increased in PMA-treated cells. Since GlcNAc T-III is the enzyme responsible for the synthesis of bis-GlcNAc, the change of bis-GlcNAc was consistent with the alteration of GlcNAc T-III activities in ATRA- and PMA-treated cells. Core alpha-1,6 fucose of surface N-glycans decreased during differentiation which was determined by AAL affinity chromatography. This was also compatible with the reduced activity of alpha-1,6 Fuc T. the enzyme catalyzing the synthesis of core Fuc, in ATRA- and PMA-treated cells. The increase of GlcNAc T-III activity and bis-GlcNAc content in N-glycans may be related to cell adhesion observed in monocytic differentiation.

Carbohydrate Sequence↗

An experimental comparison of different kinds of laryngeal muscle reinnervation.

In this study, we attempted to determine which method was the best for reinnervating the laryngeal adductor muscles by comparing nerve suture, nerve implantation, and nerve-muscular pedicle (NMP) transfer, as well as the length of time that could elapse after denervation and still allow for successful reinnervation with the ansa cervicalis. Reinnervation was performed in 36 dogs, at 6-, 8-, 10-, 12- and 18-month intervals after denervation via the three methods of muscle reinnervation described above. We noted some return of adduction in the cases using nerve suture before a 10-month interval after denervation, and with nerve implantation and NMP transfer before the 8-month intervals. The variable adduction was caused by reinnervation of the adductor muscles from the ansa cervicalis, as demonstrated by laryngeal spontaneous and evoked electromyography, the strength of muscle contraction, and histologic findings. Adduction was not observed in the cases after the above-mentioned intervals but partial improvement of the bulk and strength of the reinnervated vocal cord was still achieved. An analysis of the experimental results showed that nerve suture was superior to nerve implantation and the NMP technique. Little difference was noted between nerve implantation and the NMP technique.

Animals↗

Purification and cardiovascular activity of [Met1, Met5]-bradykinin from the plasma of a sturgeon (Acipenseriformes).

The sturgeons (Order Acipenseriformes) are extant representatives of a group of primitive Actinopterygian (ray-finned) fish that probably shared a common ancestor with present-day teleosts. Incubation of heat-denatured plasma from a sturgeon (a hybrid of the shovelnosed sturgeon Scaphirhynchus platorynchus and the pallid sturgeon Scaphirhynchus albus) with either trypsin or porcine pancreatic kallikrein generated bradykinin-like immunoreactivity. The primary structure of sturgeon bradykinin was established as Met-Pro-Pro-Gly-Met-Ser-Pro-Phe-Arg. This amino acid sequence contains two amino acid substitutions (Arg1 --> Met and Phe5 --> Met) compared with mammalian bradykinin. Bolus injections of synthetic sturgeon bradykinin in doses as low as 1 pmol/kg into the dorsal aorta of unanesthetized sturgeon resulted in an immediate and significant fall in arterial blood pressure with a maximum depressor response at 300 pmol/kg. Thus, the cardiovascular response of the sturgeon to bradykinin resembles more closely the response of mammals rather than the predominantly pressor response seen in teleost fish. Sturgeon bradykinin produced a strong and concentration-dependent (EC50 = 4.7 +/- 0.7 x 10(-10) M) relaxation of rings of vascular tissue from the sturgeon ventral aorta that had been pre-contracted with acetylcholine. The data indicate that sturgeon tissues are particularly responsive to native bradykinin and suggest that the kallikrein-kinin system may have evolved before the appearance of the neopterygians (gars, bowfin and teleosts).

Amino Acid Sequence↗

A gross and microscopic study of cerebral injuries accompanying maxillofacial high-velocity projectile wounding in dogs.

PURPOSE: This study investigated the pathologic characteristics and some related factors of cerebral damage associated with maxillofacial high-velocity missile wounds. MATERIALS AND METHODS: Sixty dogs, divided into two groups, were wounded randomly by steel spheres weighting 1.03 g at impact velocities of 1,400 m/s (46 dogs) and 800 m/s (14 dogs). Six dogs served as controls. The maxillofacial wounds and cerebral injuries were examined grossly. The distance between the center of wound and the base of skull and the largest diameter of the wound were measured, and the incidence of the brain injury in the two groups was compared. The cerebral specimens, dissected at 1 and 6 hours after trauma, were observed by light and electron microscopy. RESULTS: Cerebral hyperemia, contusion, spotty hemorrhage, and intracerebral hematoma were found in some of the dogs. In the 1,400 m/s velocity missile wound group, the incidence of the brain injury was 71.7% (33 of 46), and in the 800 m/s group it was 7.1% (1 of 14) on macroscopic examination. Microscopic observation showed intracerebral microhemotomas and degeneration and necrosis of the nervous cells. CONCLUSION: High-velocity projectile maxillofacial wounds can induce associated brain injury.

Animals↗

The rationale and method for constructing internal control DNA used in pertussis polymerase chain reaction.

The inclusion of an appropriate internal control DNA in polymerase chain reaction (PCR) is a rapid and simple method for the detection of PCR failure. Two PCR coamplification internal control DNAs (ICD I and ICD II) with the same primer-binding sequences as the target DNA for the detection of Bordetella pertussis and Bordetella parapertussis were produced using an overlap extension technique and a PCR MIMIC construction kit, respectively. The ICD II was further evaluated in a prospective clinical study in 360 patients with a clinical diagnosis of pertussis. From 360 nasopharyngeal swabs the internal control was positive in 318 (88%) samples, but was negative in 42 (12%). After phenol-chloroform extraction an additional 10 internal controls became positive. For the detection of PCR failure, the use of internal control DNA is highly recommended for PCR-based identification of B. pertussis and B. parapertussis organisms from nasopharyngeal swabs and aspirates.

Bordetella↗

Nitric oxide as an autocrine regulator of sodium currents in baroreceptor neurons.

Arterial baroreceptors are mechanosensitive nerve endings in the aortic arch and carotid sinus that play a critical role in acute regulation of arterial blood pressure. A previous study has shown that nitric oxide (NO) or NO-related species suppress action potential discharge of baroreceptors. In the present study, we investigated the effects of NO on Na+ currents of isolated baroreceptor neurons in culture. Exogenous NO donors inhibited both tetrodotoxin (TTX) -sensitive and -insensitive Na+ currents. The inhibition was not mediated by cGMP but by NO interaction with channel thiols. Acute inhibition of NO synthase increased the Na+ currents. NO scavengers (hemoglobin and ferrous diethyldithiocarbamate) increased Na+ currents before but not after inhibition of NO synthase. Furthermore, NO production in the neuronal cultures was detected by chemiluminescence and immunoreactivity to the neuronal isoform of NO synthase was identified in fluorescently identified baroreceptor neurons. These results indicate that NO/NO-related species function as autocrine regulators of Na+ currents in baroreceptor neurons. Modulation of Na+ channels may represent a novel response to NO.

Animals↗

Is functional dyspepsia just a subset of the irritable bowel syndrome?

To determine whether functional dyspepsia and irritable bowel syndrome are different entities, epidemiological data, factor analysis studies, physiological data and associated psychological symptoms were reviewed. Between 30% and 60% of patients with either diagnosis also meet the criteria for the other diagnosis, a level greater than expected to occur by chance but not sufficient to infer an identity. Most factor analysis studies identify independent clusters of symptoms corresponding to functional dyspepsia and irritable bowel syndrome. Visceral hypersensitivity is seen throughout the gastrointestinal tract in both disorders, but the motility patterns seen in association with functional dyspepsia (principally antral hypomotility and delayed gastric emptying) differ from the motility patterns seen in irritable bowel syndrome. Psychological symptoms are similar in these two disorders but are not believed to be aetiological for either of them. Thus, based on a factor analysis of gastrointestinal symptoms and differences in intestinal motility, functional dyspepsia and irritable bowel syndrome appear to be different entities.

Colonic Diseases, Functional↗

Selected symptoms associated with sexual and physical abuse history among female patients with gastrointestinal disorders: the impact on subsequent health care visits.

BACKGROUND: Despite a growing literature pointing to the deleterious health effects of sexual and physical abuse history, few studies provide evidence about which medical symptoms are most affected. The aim of this paper is to determine the impact of sexual and physical abuse history on a selected set of medical symptoms, and to test how such abuse, medical symptoms and functional disability may affect subsequent health care visits. METHODS: We studied 239 women from a referral-based gastroenterology clinic; follow-up data were available on 196 of these women. All women were interviewed about sexual and physical abuse history. RESULTS: Women with abuse history, particularly those with severe abuse, were much more likely to report somatic symptoms related to panic (e.g. palpitations, numbness, shortness of breath), depression (e.g. difficulty sleeping, loss of appetite), musculoskeletal disorders (e.g. headaches, muscle aches), genito-urinary disorders (e.g. vaginal discharge, pelvic pain, painful intercourse), skin disturbance (e.g. rash) and respiratory illness (e.g. stuffy nose). Furthermore, we found that the severity of abuse history, somatic symptoms and functional disability predicted 30% of the variance in health care visits during the subsequent year, and that the effect of abuse severity on visits was explained by abused women having more somatic symptoms and functional disability. CONCLUSIONS: Patients' reports of abuse history, somatic symptoms and functional disability appear to be important factors in explaining the number of health care visits among a clinic sample of women with gastrointestinal disorders.

Adolescent↗

Mechanical alterations in smooth muscle from mice lacking desmin.

Mice with a null mutation introduced in the desmin gene were used to study the mechanical role of intermediate filaments in smooth muscle cells. Vas deferens (VD), urinary bladder (UB) and portal vein (PV) preparations were obtained from adult animals lacking desmin (Des -/-) and from age- and weight-matched wild-type animals (Des +/+). Active force per cross-sectional area was decreased in the smooth muscle of the Des -/- compared with Des +/+ mice (VD to 42%; UB to 34%). Quantitative gel electrophoresis suggests a marginally lower cellular content of myosin, but the organization of the contractile apparatus appeared unchanged by electron microscopy. A similar reduction in stress was measured in Des -/- skinned fibres showing that altered activation mechanisms were not involved. The results indicate that the reduced active force is caused by low intrinsic force generation of the contractile filaments or subtle modifications in the coupling between the contractile elements and the cytoskeleton. The relationship between length and passive stress was less steep in the Des -/- samples and a second length force curve after maximal extension revealed a loss of passive stress. The maximal shortening velocity was reduced in Des -/- skinned VD and UB preparations by approximately 25-40%. This was associated with an increased relative content of the basic essential myosin light chain, suggesting that alterations in the contractile system towards a slower, more economical muscle had occurred. PV preparations showed no difference in mechanical properties in Des +/+ and Des -/- animals, a result that was consistent with the predominance of vimentin instead of desmin in this vascular tissue. In conclusion, the results show that, although intermediate filaments in smooth muscle are not required for force generation or maintenance of passive tension, they have a role in cellular transmission of both active and passive force.

Actins↗

The ltk gene family encodes novel receptor-like kinases with temporal expression in developing maize endosperm.

We describe the isolation and characterization of maize cDNAs that are transcribed from a small gene family and encode a novel group of receptor-like kinases (RLKs). The distinctive extracellular domain of these novel RLKs includes a unique number and arrangement of leucine-rich repeats (LRRs), a proline-rich region (PRR), a putative protein degradation target sequence (PEST), and a serine-rich region (SRR). The intracellular domain contains a putative serine/threonine protein kinase. To distinguish them from other reported RLKs, these novel RLKs were termed leucine-rich repeat transmembrane protein kinases (LTKs). Based on analysis of available deduced protein sequences, LTK1 and LTK2 were predicted to be 92.1% identical, while LTK2 and LTK3 were predicted to be 97.5% identical. Though the three LTK proteins showed high homology, the region that most distinguished LTK1 from LTK2 and LTK3 was found in the extracellular domain, in the SRR. To differentiate between expression of the individual ltk genes, we used the reverse transcriptase polymerase chain reaction (RT-PCR) in combination with restriction enzyme analysis. While ltk1 transcripts were constantly present in all tissues tested, ltk2 and ltk3 transcripts were only detected in the endosperm. Furthermore, transcript levels for both ltk1 and ltk2 showed modulation during endosperm development, peaking at 20 days after pollination. These results suggest that members of the ltk gene family mediate signals associated with seed development and maturation.

Amino Acid Sequence↗