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Biomedical subjects

J Mao

Publications and source records attributed to J Mao.

At least 91 records · Page 5Linked to original sources

Molecular characterization, sequence analysis, and taxonomic position of newly isolated fish iridoviruses.

Within the past decade, iridoviruses have been identified as the causative agents of systemic disease in a variety of commercially and recreationally important fish. Here we examine nine iridoviruses from fish, reptiles, and amphibians and demonstrate that all isolates were more similar to frog virus 3, the type species of the genus Ranavirus, than to lymphocystis disease virus, the type species of the genus Lymphocystivirus. Comparison of viral protein synthesis profiles, restriction endonuclease digestion patterns, and the amino acid sequence of the major capsid protein indicated that iridoviruses isolated from the same geographic region were similar, if not identical, whereas viruses from different areas were distinct. Moreover, using primers complementary to the conserved major capsid protein, we found that both PCR and RT-PCR successfully amplified virus-specific nucleic acid from all nine isolates. These studies demonstrate that the piscine iridoviruses examined here were members of the genus Ranavirus, and suggest that surveys of pathogenic "fish viruses" may need to include neighboring amphibian and reptilian populations. In addition, the results indicate that PCR readily identified vertebrate iridoviruses and suggest that PCR will be useful in the diagnosis of fish disease.

Amino Acid Sequence↗

The correction of Class II, division 1 malocclusion with bionator headgear combination appliance.

The purpose of this study was to evaluate skeletal and dental effects of bionator headgear combination appliances on patients in development period with Class II, division 1 malocclusion. The comparison of computerized X-ray cephalometric measurements between the 26 treated children and 26 untreated children was made. The results showed that ANB angle was significantly reduced and horizontal mandibular growth development tended to be normal in the treated group. It was suggested that the bionator headgear combination appliance can restrain the maxillary growth early and promote the forward mandibular growth which contribute the functional jaws correction.

Activator Appliances↗

The inhibition of nitric oxide-activated poly(ADP-ribose) synthetase attenuates transsynaptic alteration of spinal cord dorsal horn neurons and neuropathic pain in the rat.

Transsynaptic alteration of spinal cord dorsal horn neurons characterized by hyperchromatosis of cytoplasm and nucleoplasm (so-called 'dark' neurons) occurs in a rat model of neuropathic pain induced by chronic constriction injury (CCI) of the common sciatic nerve. The incidence of dark neurons in CCI rats has been proposed to be mediated by glutamate-induced neurotoxicity. In the present study, we examined whether the inhibition of the nitric oxide (NO)-activated poly(ADP-ribose) synthetase (PARS), a nuclear enzyme critical to glutamate-induced neurotoxicity, would both reduce the incidence of dark neurons and attenuate behavioral manifestations of neuropathic pain in CCI rats. Dark neurons were observed bilaterally (with ipsilateral predominance) within the spinal cord dorsal horn, particularly in laminae I-II, of rats 8 days after unilateral sciatic nerve ligation as compared to sham operated rats. The number of dark neurons in the dorsal horn was dose-dependently reduced in CCI rats receiving once daily intrathecal (i.t.) treatment with the PARS inhibitor benzamide (200 or 400 nmol, but not 100 nmol benzamide or saline) for 7 days. Consistent with the histological improvement, thermal hyperalgesia, mechanical hyperalgesia, and low threshold mechano-allodynia also were reliably reduced in CCI rats treated with either 200 or 400 nmol benzamide. Neither dark neurons nor neuropathic pain behaviors were reliably affected by i.t. administration of either 800 nmol novobiocin (a mono(ADP-ribose) synthetase) or 800 nmol benzoic acid (the backbone structure of benzamide), indicating a selective effect of benzamide. Intrathecal treatment with an NO synthase inhibitor NG-nitro-L-arginine methyl ester (40 nmol, but not its inactive D-isomer) utilizing the same benzamide treatment regimen resulted in similar reductions of both dark neurons and neuropathic pain behaviors in CCI rats. These results provide, for the first time, in vivo evidence indicating that benzamide is neuroprotective and that the PARS-mediated transsynaptic alteration of spinal cord dorsal horn neurons contributes to behavioral manifestations of neuropathic pain in CCI rats. These observations may have general implications beyond treatment of neuropathic pain in that PARS-mediated neuronal alterations may play a significant role in glutamate-mediated neurotoxicity under many other circumstances.

ADP Ribose Transferases↗

Expression, purification and characterization of the rat luteal 20 alpha-hydroxysteroid dehydrogenase.

The enzyme, rat ovarian 20 alpha-hydroxysteroid dehydrogenase (20 alpha HSD), plays a central role in luteolysis and parturition. It catalyzes the reduction of progesterone, leading to the formation of progestationally inactive steroid, 20 alpha-hydroxypregn-4-ene-3-one (20 alpha-hydroxyprogesterone). Recently, we reported the cloning, sequencing, and deduced amino acid sequence of the rat luteal 20 alpha HSD. To further investigate whether phosphorylation and/or glycosylation affect the activity of 20 alpha HSD and to study its kinetic and biochemical properties, we established both bacterial and insect expression systems for obtaining large quantities of enzyme. The recombinant (rec) 20 alpha HSD expressed as glutathione-S-transferase-20 alpha HSD fusion protein was purified from bacterial lysates by affinity binding to glutathione-Sepharose beads followed by thrombin digestion, whereas the rec enzyme expressed in baculovirus-insect cell system was purified to apparent homogeneity by ion exchange chromatography, followed by dye affinity chromatographies. Both rec preparations of 20 alpha HSD demonstrated a single polypeptide chain of 37 kDa with similar K(m) values for 20 alpha-hydroxyprogesterone and NADP, although the corresponding maximum velocity values were slightly lower for the rec 20 alpha HSD expressed in the insect cells. The rec 20 alpha-HSD showed preference for progesterone/20 alpha-hydroxyprogesterone. 17 alpha-Hydroxyprogesterone was only 30% as effective. The enzyme also used various substrates specific for aldo-keto reductases, although with much less efficiency. The rec enzyme preparations showed an absolute requirement for NADP(H). In vitro phosphorylation of rec bacterial enzyme with either protein kinase A or protein kinase C had no demonstrable effect on its activity. Finally, no differences in enzyme activity were noted between glycosylated (expressed in insect cells) and nonglycosylated (expressed in bacteria) forms of the enzyme. In conclusion, these studies demonstrate that rat luteal 20 alpha HSD can be prepared in large amounts from either bacterial or insect expression systems in a catalytically active form. Indirect evidence also suggests that the catalytic activity of 20 alpha HSD may be independent of phosphorylation and glycosylation states of the enzyme protein, i.e. posttranslational modification of 20 alpha HSD may not be required for the maximal expression of enzyme activity.

20-Hydroxysteroid Dehydrogenases↗

The t(10;11)(p13;q14) in the U937 cell line results in the fusion of the AF10 gene and CALM, encoding a new member of the AP-3 clathrin assembly protein family.

The translocation t(10;11)(p13;q14) is a recurring chromosomal abnormality that has been observed in patients with acute lymphoblastic leukemia as well as acute myeloid leukemia. We have recently reported that the monocytic cell line U937 has a t(10;11)(p13;q14) translocation. Using a combination of positional cloning and candidate gene approach, we cloned the breakpoint and were able to show that AF10 is fused to a novel gene that we named CALM (Clathrin Assembly Lymphoid Myeloid leukemia gene) located at 11q14. AF10, a putative transcription factor, had recently been cloned as one of the fusion partners of MLL. CALM has a very high homology in its N-terminal third to the murine ap-3 gene which is one of the clathrin assembly proteins. The N-terminal region of ap-3 has been shown to bind to clathrin and to have a high-affinity binding site for phosphoinositols. The identification of the CALM/AF10 fusion gene in the widely used U937 cell line will contribute to our understanding of the malignant phenotype of this line.

Adaptor Proteins, Vesicular Transport↗

Cloning, sequence analysis, and expression of the major capsid protein of the iridovirus frog virus 3.

The nucleotide sequence of the gene encoding in the major capsid protein (MCP) of frog virus 3 (FV3) has been determined and compared to other iridovirus capsid genes. Nucleotide sequence and S1 nuclease analysis showed that the FV3 MCP gene encoded a transcript of 1452 nucleotides containing a 12 nucleotide AU-rich 5' nontranslated region (NTR) and a 50-nucleotide 3' NTR whose terminus was predicted to fold into a hairpin of moderate stability. An open reading frame initiating from the the 5'-most AUG codon encoded a protein of 463 amino acids with a predicted molecular weight of 49,860. Expression of the putative FV3 MCP gene in vitro confirmed that it encoded the major capsid protein of FV3 and supported the suggestion that translation initiated at AUG-1. Pairwise amino acid alignments detected a high degree of sequence identity between the FV3 MCP and other iridoviruses. These results indicate that iridoviruses possess an evolutionarily related major capsid protein and provide information useful not only in studies of viral gene expression, but also in characterizing newly isolated iridoviruses.

Amino Acid Sequence↗

Identification and molecular confirmation of a small chromosome 10q duplication [dir dup(10)(q24.2-->q24.3)] inherited from a mother mosaic for the abnormality.

We describe a family in which two siblings exhibited developmental delay, reduced muscle tone and mild muscle weakness. Cytogenetic evaluation demonstrated that both children had a tandem duplication of a small portion of the long arm of chromosome 10 [46,XX or XY,dir dup(10)(q24.2-->q24.3)], inherited from their clinically normal mother, who was found to be mosaic for the duplicated chromosome 10. Fluorescence in situ hybridization approaches, including total chromosome painting and the use of regional specific cosmid probes, were used to confirm the chromosome 10q origin of the duplicated material. This is the smallest confirmed duplication of this portion of chromosome 10 reported to date.

Child↗

Power efficient on-resonance saturation pulses for magnetization transfer in magnetic resonance imaging.

A family of new on-resonance saturation pulses for magnetization transfer in MRI is proposed. These pulses can be represented as a product of a shaped function and a cosine function. The shaped function can have many different forms, one of which is a Gaussian function. The experimental results on a 1.0 T whole body scanner show that the new on-resonance pulses are more efficient for magnetization transfer than either on-resonance binomial sequence pulses or off-resonance Gaussian pulses at the same power level.

Humans↗

Vaginal hysterectomy with and without morcellation: the University of New Mexico hospital's experience.

OBJECTIVE: To compare intraoperative and postoperative complications of vaginal hysterectomy with and without morcellation. METHODS: Between August 1990 and January 1995, 345 women underwent vaginal hysterectomy without laparoscopic assistance at the University of New Mexico and associated hospitals. Medical records available for 340 of these women were reviewed for demographic data, perioperative complications, change in hematocrit, duration of procedure, hospital stay, and uterine size and weight. Some of the records were incomplete with respect to uterine size, uterine weight and hospital stay. The subjects were divided into two groups, 281 who had vaginal hysterectomies without morcellation and 59 who had vaginal hysterectomies with morcellation. The perioperative courses of the two groups were compared. RESULTS: Although women undergoing morcellation were significantly older (mean 44 versus 41 years, P = .004), there were no statistically significant differences in other surgical or anesthetic risk factors, including parity, obesity, hormonal status, and preexisting medical disease. Mean uterine weight was significantly greater in those undergoing morcellation (222 versus 110 g, P < .001). Morcellation significantly increased the duration of the procedure (mean 140 versus 168 minutes, P = .004) but did not increase the duration of hospitalization or rate of perioperative complications (18.6 versus 17.4%, odds ratio 1.09, 95% confidence interval 0.49-2.35). CONCLUSION: Uterine morcellation at the time of vaginal hysterectomy is safe and facilitates the vaginal removal of moderately enlarged and well-supported uteri without increasing perioperative morbidity.

Adult↗

Oral administration of dextromethorphan prevents the development of morphine tolerance and dependence in rats.

Combined oral administration of morphine sulfate (MS) and the over-the-counter antitussive drug and N-methyl-D-aspartate receptor antagonist dextromethorphan (DM) prevented the development of tolerance to the antinociceptive effects of MS (15, 24, or 32 mg/kg) in rats. This combined oral treatment regimen also attenuated signs of naloxone-precipitated physical dependence on morphine in the same rats. A wide range of ratios of MS to DM (2:1, 1:1, and 1:2) were effective for preventing the development of morphine tolerance and dependence. In addition, we provide evidence that under certain circumstances DM increases the acute antinociceptive effects of MS. All of these results indicate that oral treatment that combines DM with opiate analgesics may be a powerful approach for simultaneously preventing opiate tolerance and dependence and enhancing analgesia in humans.

Administration, Oral↗

A common region of 10p deleted in DiGeorge and velocardiofacial syndromes.

DiGeorge (DGS, MIM 188400) and velocardiofacial (VCFS, MIM 192430) syndromes may present many clinical problems including cardiac defects, hypoparathyroidism, T-cell immunodeficiency and facial dysmorphism. They are frequently associated with deletions within 22q11.2, but a number of cases have no detectable molecular defect of this region. A number of single case reports with deletions of 10p suggest genetic heterogeneity of DGS. Here we compare the regions of hemizygosity in four patients with terminal deletions of 10p (one patient diagnosed as having hypoparathyroidism and three as DGS) and one patient with a large interstitial deletion (diagnosed as VCFS). Fluorescence in situ hybridization (FISH) analysis demonstrates that these patients have overlapping deletions at the 10p13/10p14 boundary. A YAC contig spanning the shortest region of deletion overlap (SRO) has been assembled, and allows the size of SRO to be approximated to 2 Mb. As with deletions of 22q11, phenotypes vary considerably between affected patients. These results strongly support the hypothesis that haploinsufficiency of a gene or genes within 10p (the DGSII locus) can cause the DGS/VCFS spectrum of malformation.

Abnormalities, Multiple↗

SP75 is encoded by the DP87 gene and belongs to a family of modular Dictyostelium discoideum outer layer spore coat proteins.

Highly purified spore coats of Dictyostelium discoideum each contained about 5 x 10(6) protein molecules as determined by amino acid composition analysis. By two-dimensional gel electrophoresis the coats were found to contain nine major-abundance and numerous minor protein species, most of which were highly enriched relative to the adjacent interspore matrix. Protein was nearly quantitatively eluted by denaturants and 2-mercaptoethanol, showing that it was not irreversibly cross-linked. Because a reducing agent is required together with denaturants to elute most proteins if their free thiol groups have been prealkylated, it was concluded that the D. discoideum spore coat proteins are disulfide cross-linked into the matrix. One major coat protein, SP75, was partially sequenced and found to be encoded by the previously identified DP87 gene; this finding was supported by additional physical, genetic, biochemical and microscopic evidence. The five major proteins for which genes have been cloned were associated with the outer layer of the coat. In coats missing one or more of four of these proteins as a result of gene disruption, there were physical changes but, with one exception, the other major coat proteins appeared to be incorporated normally. Sequence analysis showed that these five outer layer coat proteins are homologous and consist of alternating sequence motifs related to epithelial mucin repeats, basic proline repeats found in salivary acidic proline-rich proteins, the NH2-terminal subdomain of epidermal growth factor modules and other cysteine repeats. Based on these and other observations, outer layer coat proteins are predicted to organize indeterminately to form a cell surface microenvironment supportive of cellulose morphogenesis during spore coat formation.

Amino Acid Sequence↗

Characterization of a flow-sorted human chromosome 10 cosmid library by FISH.

Fluoresence in situ hybridization (FISH) was used to localize cosmids to regions of human chromosome 10. A total of 301 cosmids were selected randomly from a flow-sorted human chromosome 10 cosmid library constructed from human x hamster cell line 762-8A and arrayed in microtiter storage dishes. Over 70% (211/301) of the cosmids mapped to unique regions of chromosome 10. About 7% (22/301) produced multiple hybridization signals indicative of chimeric clones or sequences repeated at low copy number. Three cosmids (3/301, or 1%) hybridized to the centromeric regions of chromosome 10 and one or more other human chromosomes. About 19% (59/301) consisted mostly or entirely of hamster DNA inserts, and about 2% (6/301) appeared to be nonrecombinants.

Animals↗

Experience in prevention of serious complications of laparoscopic cholecystectomy.

OBJECTIVE: To study the causes and prevention of the complications of laparoscopic cholecystectomy (LC). PATIENTS AND METHODS: Based on experience with 2 428 cases, the following should be paid attention to when dissecting and separating adhesions around the gallbladder and of the Calot's triangle. The best method for the prevention of mistaking the common bile duct (CBD) for the cystic duct is to find the junction of the cystic infundibulum and duct, separate the gallbladder wall along the infundibulum, and transect the cystic duct at the junction with the infundibulum. If dense adhesions around the gallbladder or of the Calot's triangle are met with, LC should be abandoned and open the cholecystectomy (OC) should be used instead. In separating the Calot's triangle, blunt dissection should be used to avoid burning the extrahepatic bile duct (EHBD), and blind hemostasis should be avoided. If the cystic artery lies in the upper part and the back of the cystic duct, the cystic duct should be dissected out, clipped and cut first, then the cystic artery be dealt with. If the cystic artery is in the front part of the pedicle of the gallbladder, the artery should be separated, clipped and cut first. Injury to the adjacent organs may be avoided by using electric coagulating hook correctly and avoiding accidental damage to the viscera, and keeping from viscera injury due to current chemotaxis in the closed cavity of the body. RESULTS: A total of 2427 patients were cured. One patient died of frequent episodes of supraventricular tachycardia and pneumonia on the 21st day after LC. CONCLUSION: If LC surgeons follow the above said principles of LC technique. LC is very safe for patients with benign diseases of the gallbladder.

Adolescent↗

The coiled coil stem loop miniprotein as a presentation scaffold.

The coiled coil helical dimer found in naturally occurring proteins is conformationally stable and can tolerate significant sequence variation on the solvent-exposed surfaces of the helices. We are interested in exploring the use of a de novo designed coiled coil stem loop miniprotein (CCSL) as a template for presenting (1) helical and loop sequences from heterologous proteins and (2) constrained libraries of peptides. Towards this end, we synthesized a 56 residue prototype CCSL and verified its structure by extensive biophysical characterization. CCSL variants with altered sequences in the solvent-exposed helical positions were found to fold similarly to the prototype design. Based on the results with the CCSL produced by peptide synthesis, we assembled a synthetic cDNA for the CCSL prototype and expressed the CCSL miniprotein on filamentous phage. This genetic construction can be used to introduce random peptide libraries into regions within the scaffold loop and helices in order to identify key side chains of native proteins involved in binding, to establish structural models for their pharmacophores and to identify novel peptide recognition mimics of macromolecular ligands and their receptors.

Amino Acid Sequence↗