Biomedical subjects

N Zhao

Publications and source records attributed to N Zhao.

At least 37 records · Page 2Linked to original sources

PubMed2000

cDNA cloning, expression pattern, genomic structure and chromosomal location of RAB6KIFL, a human kinesin-like gene.

Kinesin-like proteins are microtubule-associated motors that play important roles in intracellular transport and cell division. We report here the characterization of a new human kinesin-like protein, Rabkinesin6 (RAB6KIFL). The composite cDNA sequence is 2957bp, and encodes a protein of 890 amino acids with a predicted molecular weight of 100kDa. It has high homology (93% similarity) with the mouse kinesin-like protein, Rab6kifl, indicating that it is the human homologue of the mouse gene. RAB6KIFL has all of the structural characteristics required to function as a microtubule-associated motor. Unlike the mouse gene which is ubiquitously expressed, RAB6KIFL expression appears to be tissue specific. It is widely expressed in fetal tissues, abundantly expressed in adult thymus, bone marrow and testis, and is expressed at low levels in heart, placenta and spleen. The RAB6KIFL gene is mapped to human chromosome 5, band q31, spans approximately 8.5kb of genomic DNA, and contains 19 exons. RAB6KIFL maps within the smallest commonly deleted segment in myeloid leukemias characterized by a deletion of 5q; however, we detected no mutations of RAB6KIFL in malignant myeloid disorders with loss of 5q. The description of this human kinesin-like protein may provide a better understanding of the diversity of this large family of proteins.

Acute Disease↗

PubMed2000

Photochemistry of substituted benzyldiphenylphosphine oxides

Photochemical reactions (lambda irr = 254 nm) of substituted benzyldiphenylphosphine oxides 1a-e have been investigated in benzene and acetonitrile. alpha-Cleavage from the singlet excited state is proposed as the primary process, and products formed both before and after escape of the primary intermediates from the solvent cage result. Radicals 2 and 3 are observed by nanosecond flash photolysis following excitation of 1a at 266 nm in acetonitrile. Acetone sensitization of 1a,b fails to improve the efficiency of product formation. The quantum yields of disappearance of 1a,b are unaffected by O2, and mechanisms for product formation from 1a in the presence of oxygen are proposed.

Journal Article↗

PubMed2000

Spectroscopic study of trypsin, heat and triton X-100-induced denaturation of the chlorophyll-binding protein CP43.

Trypsin-, heat- and Triton X-100-induced denaturation of CP43, the core antenna complex of photosystem II purified from spinach, has been investigated using absorption, fluorescence and circular dichroism spectroscopy. Triton X-100 was found to bring about considerable dissolution of pigments from the protein to the monomeric state in solution and destruction of the interactions among the chlorophyll, carotene and protein. Heat induced significant unfolding of the protein secondary structure and loss of excitonic interactions of the pigments, but no apparent dissolution of the pigments from CP43. Trypsin caused structural changes in the extrinsic part of the protein but no change of the native state of the pigments. Trypsin, heat and Triton X-100 treatments increased the light sensitivity of chlorophyll in CP43 to different extents. The results suggest that the protein and beta-carotene can protect the chlorophyll from light-induced destruction in CP43.

Binding Sites↗

PubMed2000

Analysis of libraries encoded with GC tags: compound elution, tag decode analysis, and statistical sampling analysis.

Libraries encoded with electrophoric tags present a unique challenge with respect to library quality control and characterization. Libraries are prepared on Tentagel resin in 200-fold redundancy wherein each resin particle contains one compound per one tag set. The amount of compound present on the bead is ca. 200-500 pmole while tag levels are estimated at 0.5-1 pmol/bead. Several quality control protocols have been developed in order to accurately estimate bead yield and purity for the entire library, ensure high tag fidelity, and to determine the overall performance of individual synthons. This review provides a unique, collective portrait of Pharmacopeia's approach in assessing the quality of libraries prepared using its molecular encoding technology.

Chromatography, Liquid↗

PubMed2000

Npt2 gene disruption confers resistance to the inhibitory action of parathyroid hormone on renal sodium-phosphate cotransport.

PTH inhibition of renal sodium-phosphate (Na-Pi) cotransport is associated with the endocytic retrieval of the type II Na-Pi cotransporter, Npt2, from the renal brush border membrane into the late endosomal/lysosomal compartment. The aim of the present study was to determine whether mice homozygous for the disrupted Npt2 gene (Npt2-/-) exhibit decreased renal Pi reabsorption in response to PTH. We demonstrate that PTH has no effect on the serum Pi concentration, fractional excretion of Pi, or Na-dependent Pi transport in renal brush border membrane vesicles in Npt2-/- mice. In contrast, PTH elicits a fall in the serum Pi concentration, an increase in urinary Pi excretion, a decrease in brush border membrane Na-Pi cotransport, and a corresponding reduction in the relative abundance of Npt2 protein in wild-type mice (Npt2+/+). Both Npt2-/- and Npt2+/+ mice exhibit a significant rise in the urinary cAMP/creatinine ratio in response to PTH, indicating that generalized resistance to PTH cannot account for the absence of the PTH response in Npt2-/- mice. In addition, we demonstrate that Pi-depleted normal mice respond to PTH with a decrease in renal brush border membrane Na-Pi cotransport and Npt2 protein, indicating that Pi deficiency per se does not account for PTH resistance in Npt2-/- mice. Taken together, our data provide compelling evidence that Npt2 gene expression is crucial for PTH effects on renal Pi handling.

Animals↗

PubMed2000

Susceptibility gene for familial acute myeloid leukemia associated with loss of 5q and/or 7q is not localized on the commonly deleted portion of 5q.

The molecular mechanism for the occurrence of leukemia in multiple members of a family has not been fully elucidated but data support the contribution of highly penetrant mutations in leukemia susceptibility genes. We have investigated the genetic etiology of an unusual three-generation family with apparent autosomal dominant transmission of acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) accompanied by somatic loss of the long arm of chromosome 5 and/or loss of heterozygosity (LOH) analysis and fluorescence in situ hybridization (FISH) of leukemia cells have been performed, confirming acquired hemi- and homozygous deletion of the long arm of chromosome 5. However, the chromosome lost in the observed LOH event is from the affected parent, in contradiction to the expectation for a two-hit hypothesis involving a tumor suppressor gene. Furthermore, genetic linkage has been performed at 5q31-33 as well as other loci (21q22 and 16q21-23.2) previously implicated in familial leukemia. In this family, linkage analysis excludes loci at 5q31-33 and 21q22, but localization to 16q21-23.2 cannot be excluded. We observed a maximum multipoint LOD score of 1.19 between marker D16S265 and D16S503 at 16q22 (P = 0.03), suggesting possible linkage to this locus. Considering this family and the previous 16q-linked family together, the linkage of a leukemia susceptibility gene to 16q22 achieved an LOD score of 3.63 at D16S265 with theta = 0. Thus, somatic deletion of the long arm of chromosome 5 appears as a necessary but surprisingly noncausative event for onset of AML and MDS in this family, thereby confirming a multistep etiology in which chromosome 5 plays an important secondary role.

Acute Disease↗

PubMed1999

Two proteins share immunological epitopes on the tumor-associated antigen 17-1A.

The mouse monoclonal antibody (mAb) 17-1A which recognizes the tumor-associated antigen 17-1A (also called EGP-40 or EpCAM) was successfully used in adjuvant therapy for colorectal carcinoma. In the 17-1A antigen analysis, we isolated not only a protein of 33 kDa (P33) which was reported as the tumor associated antigen 17-1A, but also a protein of 65 kDa (P65) using affinity chromatography from cell lysates of HCT, and another protein of 50 kDa (P50) from lysates of human colorectal tumor tissues. The mAbs 17-1A and M79 (mAb M79 recognizes a different epitope on the 17-1A antigen) both could bind P33 and P50, but only M79 bound to P65 in an enzyme-linked immunosorbant assay (ELISA). These results indicate that P33 and P50 share at least two epitopes, and a common immunological epitope exists among P33, P50 and P65, suggesting that the two new proteins (P50 and P65) are related to the tumor-associated antigen 17-1A.

Animals↗

PubMed1999

TTID: A novel gene at 5q31 encoding a protein with titin-like features.

A deletion of the long arm of chromosome 5 is a recurring abnormality in malignant myeloid disorders. In previous studies, we identified an approximately 1-Mb segment in 5q31 that was deleted in all patients examined. As part of a positional cloning project to identify transcribed sequences in this region, we identified and characterized the TTID gene. This gene contains 10 exons that extend over 19 kb. The composite cDNA is approximately 2.3 kb and encodes a protein of 498 amino acids, with a predicted molecular mass of 55 kDa. The C-terminal half of this putative protein contains an internally repeated domain of 43 amino acids, which resembles the N-terminal half of an immunoglobulin domain from the immense skeletal muscle protein titin. The TTID gene is expressed in multiple muscle tissue types as well as in thyroid gland and bone marrow. We evaluated the gene as a candidate tumor suppressor gene by searching for mutations in malignant myeloid disorders with abnormalities of chromosome 5. However, we detected no inactivating mutations. A single nucleotide change (G to A) was identified at nucleotide position 1889 in the untranslated region of the mRNA, which may represent a polymorphism. Therefore, TTID is unlikely to be the candidate tumor suppressor gene involved in malignant myeloid disorders.

Amino Acid Sequence↗

PubMed1999

Thermal conformational changes of bovine fibrinogen by differential scanning calorimetry and circular dichroism.

The thermal denaturation of bovine fibrinogen has been investigated using differential scanning calorimetry (DSC) and circular dichroism (CD) spectroscopy. Differential scanning calorimetry measurements were carried out while changing the scan-rate. The transition at 57 degrees C was found to be irreversible and highly scan-rate dependent, suggesting that the denaturation is, at least in part, under kinetic control. The secondary structural changes at various temperatures were monitored by far-ultraviolet CD spectroscopy. These results show that the DSC transition for the thermal denaturation of bovine fibrinogen can be interpreted in terms of a kinetic process, N --> F, where k is a first-order kinetic constant that changes with temperature according to the Arrhenius equation. An important transition peak was observed at 78.8 degrees C which is attributed to the C-terminal parts of the Aalpha chains of fibrinogen.

Animals↗

PubMed1999

Double-blind, randomized study of nalmefene and naloxone in emergency department patients with suspected narcotic overdose.

STUDY OBJECTIVES: To compare the efficacy, safety, and withdrawal symptoms in emergency department patients with suspected narcotic overdose treated with nalmefene, an opioid antagonist with a 4- to 10-hour duration of action, with those treated with naloxone. METHODS: Adults in 9 centers who would otherwise receive naloxone for altered consciousness levels were randomly assigned to receive intravenous study drug (1 mg nalmefene, or 2 mg nalmefene or 2 mg naloxone, double-blinded) every 5 minutes as needed for up to 4 doses in a 4-hour study. Outcomes were 20-minute and 4-hour posttreatment changes in respiratory rates, Neurobehavioral Assessment Scale scores, Opioid Withdrawal Scale scores, and incidences of adverse events. RESULTS: Opioid positivity was recorded for 30 of 63 (1-mg nalmefene), 23 of 55 (2-mg nalmefene), and 24 of 58 (naloxone) cases, 75% of whom also had nonopioid central nervous system depressants. Most patients received only 1 dose of study drug. Similar, clinically meaningful improvements in respiratory rates and Neurobehavioral Assessment Scale scores were seen with all treatments. No statistical differences in efficacy or withdrawal outcomes were seen between treatment groups, and no significant overall time-treatment interactions occurred, in either the entire patient group or among opioid-positive cases (P >.21, all comparisons). Adverse events occurred in 30.9% (2 mg nalmefene), 15.9% (1 mg nalmefene), and 15.5% (naloxone) of patients (P >.08); none were associated with morbidity. CONCLUSION: In this study of patients with varied potential causes of altered consciousness, nalmefene (1 mg and 2 mg) and naloxone (2 mg) appeared to be efficacious, safe, and to yield similar clinical outcomes.

Adult↗

PubMed1999

Effects of chlorophyll availability on phycobilisomes in Synechocystis sp. PCC 6803.

Inactivation of the chlL gene in Synechocystis sp. PCC 6803 resulted in negligible chlorophyll content when the mutant was grown in darkness. Upon phycocyanin excitation at 580 nm, the 77K fluorescence spectrum of dark-grown cells showed three peaks at 648 nm, 665 nm, and 685 nm, this last being the largest. This reflects the functional presence of major components of phycobilisomes, including phycocyanin, allophycocyanin, and the terminal emitter, and efficient energy transfer between these components. As expected, no fluorescence emission peaks corresponding to chlorophyll in the photosystems were observed. Intact phycobilisomes could be isolated from the dark-grown chlL-deletion mutant. However, the phycobilisomes had a lower efficiency of energy transfer than did those isolated from the light-grown mutant, probably because of a decreased phycobilisome stability in the absence of chlorophyll. Exposing the dark-grown chlL-deletion mutant to light triggered the biosynthesis of chlorophyll. For the first 6 h in the light, upon phycocyanin excitation at 580 nm, the 77K fluorescence emission spectrum of greening cells was identical to that of dark-grown cells that lacked significant amounts of chlorophyll. With increased chlorophyll synthesis, gradual energy transfer from phycobilisomes to the two photosystems can be demonstrated.

Bacterial Proteins↗

PubMed1999

Utility of cardiac troponin I, creatine kinase-MB(mass), myosin light chain 1, and myoglobin in the early in-hospital triage of "high risk" patients with chest pain.

OBJECTIVE: To evaluate the use of cardiac troponin I (cTnI), creatine kinase-MB(mass) (CK-MB(mass)), myosin light chain 1 (MLC 1), and myoglobin in identifying "high risk" patients with chest pain who will experience serious cardiac events (SCEs) in hospital. DESIGN: Prospective study. SETTING: University affiliated medical centre in Philadelphia, USA. PATIENTS: 208 patients with chest pain, at > 7% risk of acute myocardial infarction (MI), but without new ST segment elevation on their presenting ECG. INTERVENTIONS: cTnI, CK-MB(mass), MLC 1, and myoglobin concentrations were obtained on admission (0 hour) and at 4, 8, 16, and 24 hours. MAIN OUTCOME MEASURES: The sensitivity, specificity, positive and negative predictive value, and pre- and post-test probabilities of patients suffering an SCE in hospital were determined. SCEs included cardiac death, acute MI, cardiac arrest, life threatening cardiac arrhythmia, cardiogenic shock, and urgent coronary revascularisation. RESULTS: Admission concentrations of all markers were poor predictors of SCEs in hospital but improved substantially at subsequent timepoints. cTnI and CK-MB(mass) were consistently the most useful prognostic indicators. If both were negative at 0, 4, and 8 hours, then 99% (95% confidence interval 96% to 100%) of patients remained free from SCEs. The only SCEs not thus predicted were revascularisation procedures and associated complications. Additional tests after 8 hours, or the inclusion of additional markers, did not improve predictive accuracy further. CONCLUSIONS: Patients with high risk clinical features on admission who have negative cTnI and CK-MB(mass) concentrations at 0, 4, and 8 hours later have a favourable in-hospital prognosis and could be considered for early triage out of coronary care units.

Biomarkers↗

PubMed1999

[Growth behavior of cells of ameloblastoma and odontogenic keratocyst].

OBJECTIVE: To compare the growth behavior of epithelium and fibroblast from ameloblastoma and keratocyst and to discuss the recurrence reason for the keratocyst. METHODS: Epithelial cells and fibroblast from ameloblastoma, keratocyst, dentigerous cyst, radicular cyst and oral mucosa were cultured and observed by phase-contrast microscope. RESULTS: The epithelial cells of ameloblastoma grew faster, with an irregular area around the cell mass and some small satellite-shaped cell mass. The epithelial cells of all the cysts and mucosa grew slower with the cell masses surrounded by fibroblasts which grew in whirl manner with a round and smooth periphery. CONCLUSION: The cells from the ameloblastoma grow faster and more active, and those from the keratocysts grow like the other cysts and the normal mucosa. It is speculated that the ameloblastoma grows in an aggressive manner and it is possible that the recurrence of the keratocyst isn't primarily resulted from its growth behavior as a tumor.

Ameloblastoma↗

PubMed1999

[V-ATPase inhibitor baflomycine A1 inhibits bone resorption by osteoclast-like cells].

OBJECTIVE: In order to compare the role of osteoclast-like cells (OLCs) originating from giant cell tumor of bone and the osteoclasts (OCs) on bone resorption, as well as to study which kind of proton pump is associated with bone resorption by the OLCs and its role related. METHODS: Culture of OLCs together with the bone slices, the number of pits and the surface areas formed by OLCs was counted. The effects of F0F1-ATPase inhibitor, oligmycine V-ATPase inhibitor, baflomycine of OLCs on bone resorption were compared. RESULTS: Baflomycine are considered effective in inhibiting bone resorption but no definited effect obtained with oligmycine. CONCLUSIONS: OLCs bear the similar bone resorption mechanism as that of OCs. It's considered that OLCs and OCs are derived from the same origin.

Animals↗

PubMed1999

[Determination of sarsasapogenin in Anemarrhena asphodeloides Bunge by GC].

OBJECTIVE: To determine sarsasapogenin in Anemarrhena asphodeloides. METHOD: Chloromethane extract (1 microliter) with cholesterol as internal standard was analyzed on HP-1 column, operated at 270 degrees C with N2 as carrier gas and FID. RESULT: The method was linear within the range of 0.245-2.94 micrograms.microliter-1, with a correlation coefficient of 0.9996. The average recovery was 95.52% +/- 1.77%, CONCLUSION: The method is reproducible, rapid and sensitive.

Anemarrhena↗

PubMed1998

Cloning and mapping of ZNF231, a novel brain-specific gene encoding neuronal double zinc finger protein whose expression is enhanced in a neurodegenerative disorder, multiple system atrophy (MSA).

A novel brain-specific gene, neuronal double zinc finger protein (ZNF231), was cloned and mapped. We used the high-density cDNA filter method to analyze the gene-expression profile in brains with multiple system atrophy (MSA). MSA is a sporadic progressive neurodegenerative disease characterized clinically by cerebellar symptoms, parkinsonism, autonomic dysfunction, or their various combinations, but its pathogenesis has yet to be clarified. In total, 8300 cDNA clones were screened, and a novel gene, ZNF231, was identified, whose expression was elevated in cerebella of patients with MSA. Its transcript is approximately 16 kb long and encodes an open reading frame of 3926 amino acid residues that has several interesting motifs; two glycine-proline dipeptide repeats (aa 22-32 and aa 61-74), a pair of homologous C8 double zinc finger motifs (aa 169-226 and aa 465-521), a leucine zipper motif (aa 561-582), a SH3 domain-binding motif (aa 825-831), two nuclear targeting signals (aa 1011-1028 and aa 1071-1091), two glutamine-rich domains (aa 2428-2473 and aa 3775-3804), and a histidine-rich domain (aa 3597-3682). These features suggest that the new gene encodes a nuclear protein or transcription regulator. Northern blot and RT-PCR analyses showed that its expression is specific to the brain and apparently restricted to the neurons. Elevation of ZNF231 expression may be involved in the pathogenesis of multiple system atrophy. The gene for ZNF231 is located on chromosome 3p21.

Amino Acid Sequence↗

PubMed1998

Human CDC23: cDNA cloning, mapping to 5q31, genomic structure, and evaluation as a candidate tumor suppressor gene in myeloid leukemias.

The transition from metaphase to anaphase and exit from mitosis involve the degradation of active cyclin B-CDC2 complexes by ubiquitin-mediated proteolysis. The anaphase-promoting complex (APC) catalyzes the formation of cyclin B-ubiquitin conjugates, thereby targeting cyclin B for degradation. The APC is composed of eight proteins, including four members of a family characterized by multiple tetratricopeptide repeats (TPR). We mapped two overlapping expressed sequence tag clones within a genomic contig on human chromosome 5, band q31. A search revealed high homology to Saccharomyces cerevisiae CDC23, a TPR protein component of the APC. We have isolated the human CDC23 cDNA containing the full-length predicted open reading frame. The approximately 3.3-kb message is ubiquitously expressed and encodes a protein with 591 amino acids (MW = 68,293 Da) and 9 TPR units. The protein has 30% identity and 51% similarity to the S. cerevisiae protein. The human CDC23 gene contains 16 exons and spans approximately 31 kb. CDC23 maps within the smallest commonly deleted segment in myeloid leukemias characterized by a deletion of 5q; however, we detected no mutations of CDC23 in leukemia cells with loss of 5q. Thus, CDC23 is unlikely to be involved in the pathogenesis of myeloid leukemias characterized by abnormalities of chromosome 5.

Amino Acid Sequence↗

PubMed1998

Patterns of injury in belted and unbelted individuals presenting to a trauma center after motor vehicle crash: seat belt syndrome revisited.

STUDY OBJECTIVE: Investigators have described a "seat belt syndrome" consisting variously of injuries to the lumbar or cervical spine, abdominal contents, or all 3. In this study we sought to identify these and any other patterns of injury associated with seat belt use in patients who presented to a trauma center after a motor vehicle crash. METHODS: The charts of all patients involved in motor vehicle crashes who presented as trauma alerts to the study institution between January 1, 1991, and December 31, 1993, were retrospectively reviewed for data regarding belt use and 35 specific injuries in 7 body regions. We calculated the positive likelihood ratio of injury between belted and unbelted patients, along with 95% confidence intervals. RESULTS: We identified 1,124 patients involved in motor vehicle crashes. Of these subjects, 376 were belted and 544 unbelted; in 204 belt status was unknown. Belted patients were more likely to have sustained sternal fracture than were unbelted patients (4% versus .7%; positive likelihood ratio, 1.97; 95% confidence interval, 1.09 to 3.29) but were less likely to have sustained head injury (30.6% versus 46.0%; positive likelihood ratio, .67; 95% confidence interval, .53 to .83). We noted no statistically significant differences in the rates of other specific injuries, including cervical and lumbar fractures and the need for abdominal surgery. CONCLUSION: Severe injuries of all types occur in both belted and unbelted individuals involved in motor vehicle crashes who present to a typical trauma center. With the exception of sternal fractures, injuries previously associated with the seat belt syndrome occurred in similar proportions of belted and unbelted patients. Head injuries were less frequent. Seat belt use cannot serve as a discriminator for specific injury. A diligent search of all body regions is indicated in both belted and unbelted patients.

Abdominal Injuries↗