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Biomedical subjects

C C Wang

Publications and source records attributed to C C Wang.

At least 145 records · Page 8Linked to original sources

Inhibition of pyruvate-ferredoxin oxidoreductase gene expression in Giardia lamblia by a virus-mediated hammerhead ribozyme.

Giardia lamblia is a primitive eukaryotic microorganism that derives its metabolic energy primarily from anaerobic glycolysis. In trophozoites, pyruvate-ferredoxin oxidoreductase (PFOR) converts pyruvate to acetyl-CoA with the transfer of a pair of electrons to ferredoxin, which can then reduce metronidazole and activate it into a potent antigiardiasis agent. It is unclear, however, whether this anaerobic disposal of electrons is essential for the energy metabolism in Giardia. In the present study, cDNAs encoding hammerhead ribozyme flanked with various lengths of antisense PFOR RNA were cloned into a viral vector pC631pac derived from the genome of giardiavirus (GLV). RNA transcripts of the plasmids showed high cleavage activities on PFOR mRNA in vitro. They were introduced into GLV-infected G. lamblia trophozoites by electroporation and stablized in the transfected cells via serial passages under puromycin selection. PFOR mRNA and enzyme activity in the transfected cells were decreased by 46-60% with the ribozyme PRzS flanked with 20 nt PFOR antisense RNA on each arm and by 69-80% with the ribozyme PRzL flanked with 600 and 1500 nt PFOR antisense RNA. PRzS without the inserted ribozyme or ribozyme flanked with alcohol dehydrogenase E antisense RNA showed no effect on PFOR mRNA and activity. The ribozyme-transfected cells demonstrated significantly enhanced resistance to metronidazole and grew equally well under anaerobic and aerobic conditions. In contrast, the wild-type cells grew slightly better anaerobically than the transfectants but did not grow at all in aerobic conditions. Thus, the reduced PFOR expression enables Giardia to grow under molecular oxygen and the presence of PFOR enhances the anaerobic growth of Giardia with an increased susceptibility towards metronidazole. In addition, this study demonstrated for the first time the feasibility of using a viral RNA vector to express a ribozyme targeted at a specific mRNA in G. lamblia to reduce the expression of a specific gene.

Animals↗

Color ultrasonography: a useful technique in the identification of nuchal cord during labor.

OBJECTIVE: To compare the accuracy of intrapartum ultrasound with and without color Doppler for identification of nuchal cord displacement during labor. METHODS: 180 normal pregnant women, admitted in labor, or for induction to labor, were examined independently by two researchers, using either conventional real-time gray-scale imaging or color Doppler imaging. A repeat examination was performed by the other researcher using the other ultrasound modality. Nuchal cord displacement was classified as either negative, definite or suspicious. Nuchal cord at birth was classified as either tight or loose. RESULTS: Sixty-two (34%) cases examined using the two ultrasound imaging modalities presented with nuchal cord at delivery. The sensitivity of color Doppler was 96.8%. The accuracy of color Doppler in detecting nuchal cord during labor was significantly better (P < 0.05) than gray-scale imaging alone. The results of a restricted sequential t-test analysis of 53 un-tied pairs showed an overall preference in favor of color Doppler assessment: statistical significance (P < 0.01) was reached after 41 un-tied pairs. CONCLUSIONS: The tight and loose nuchal cord could not be distinguished by ultrasound. Color Doppler imaging can provide useful additional information to gray-scale imaging in the detection of nuchal cord displacement during labor.

Female↗

Boron neutron capture therapy of brain tumors: biodistribution, pharmacokinetics, and radiation dosimetry sodium borocaptate in patients with gliomas.

OBJECTIVE: The purpose of this study was to obtain tumor and normal brain tissue biodistribution data and pharmacokinetic profiles for sodium borocaptate (Na2B12H11SH) (BSH), a drug that has been used clinically in Europe and Japan for boron neutron capture therapy of brain tumors. The study was performed with a group of 25 patients who had preoperative diagnoses of either glioblastoma multiforme (GBM) or anaplastic astrocytoma (AA) and were candidates for debulking surgery. Nineteen of these patients were subsequently shown to have histopathologically confirmed diagnoses of GBM or AA, and they constituted the study population. METHODS: BSH (non-10B-enriched) was infused intravenously, in a 1-hour period, at doses of 15, 25, and 50 mg boron/kg body weight (corresponding to 26.5, 44.1, and 88.2 mg BSH/kg body weight, respectively) to groups of 3, 3, and 13 patients, respectively. Multiple samples of tumor tissue, brain tissue around the tumors, and normal brain tissue were obtained at either 3 to 7 or 13 to 15 hours after infusion. Blood samples for pharmacokinetic studies were obtained at times up to 120 hours after termination of the infusion. Sixteen of the patients underwent surgery at the Beijing Neurosurgical Institute and three at The Ohio State University, where all tissue samples were subsequently analyzed for boron content by direct current plasma-atomic emission spectroscopy. RESULTS: Blood boron values peaked at the end of the infusion and then decreased triexponentially during the 120-hour sampling period. At 6 hours after termination of the infusion, these values had decreased to 20.8, 29.1, and 62.6 microg/ml for boron doses of 15, 25, and 50 mg/kg body weight, respectively. For a boron dose of 50 mg/kg body weight, the maximum (mean +/- standard deviation) solid tumor boron values at 3 to 7 hours after infusion were 17.1+/-5.8 and 17.3+/-10.1 microg/g for GBMs and AAs, respectively, and the mean tumor value averaged across all samples was 11.9 microg/g for both GBMs and AAs. In contrast, the mean normal brain tissue values, averaged across all samples, were 4.6+/-5.1 and 5.5+/-3.9 microg/g and the tumor/normal brain tissue ratios were3.8 and 3.2 for patients with GBMs and AAs, respectively. The large standard deviations indicated significant heterogeneity in uptake in both tumor and normal brain tissue. Regions histopathologically classified either as a mixture of tumor and normal brain tissue or as infiltrating tumor exhibited slightly lower boron concentrations than those designated as solid tumor. After a dose of 50 mg/kg body weight, boron concentrations in blood decreased from 104 microg/ml at 2 hours to 63 microg/ml at 6 hours and concentrations in skin and muscle were 43.1 and 39.2 microg/g, respectively, during the 3- to 7-hour sampling period. CONCLUSION: When tumor, blood, and normal tissue boron concentrations were taken into account, the most favorable tumor uptake data were obtained with a boron dose of 25 mg/kg body weight, 3 to 7 hours after termination of the infusion. Although blood boron levels were high, normal brain tissue boron levels were almost always lower than tumor levels. However, tumor boron concentrations were less than those necessary for boron neutron capture therapy, and there was significant intratumoral and interpatient variability in the uptake of BSH, which would make estimation of the radiation dose delivered to the tumor very difficult. It is unlikely that intravenous administration of a single dose of BSH would result in therapeutically useful levels of boron. However, combining BSH with boronophenylalanine, the other compound that has been used clinically, and optimizing their delivery could increase tumor boron uptake and potentially improve the efficacy of boron neutron capture therapy.

Adult↗

Determinants of umbilical cord arterial 8-iso-prostaglandin F2alpha concentrations.

OBJECTIVE: To determine the concentration of 8-isoPGF(2alpha) in cord blood as a measure of oxidative stress during labour, and to compare them with other established parameters of in vivo lipid peroxidation and with the acid-base status of the newborn. METHOD: Umbilical cord arterial and venous blood samples were collected from 81 singleton term deliveries for determination of 8-isoPGF(2alpha), malondialdehyde and organic hydroperoxides. In addition, metabolites derived from the oxidative metabolism of purines during hypoxia-reoxygenation and routine cord blood of oxygen saturation, pH, pO2, pCO2, HCO3 and base excess were measured. RESULTS: Arterial concentrations of 8-isoPGF(2alpha) were significantly higher in cases with fetal distress, tight nuchal cord (P < 0.001), the umbilical coiling index, and male sex (P < 0.05) (R2 = 0.48). No correlation was found with any parameter of acid-base status. In arterial and venous blood the concentrations of organic hydroperoxides and hypoxanthine significantly correlated with the fetal nuchal cord (P < 0.001) (R2 = 0.26 and 0.16, respectively). CONCLUSION: Our findings indicate that 8-isoPGF2(alpha) in cord arterial blood is a suitable parameter to quantify a possible oxidative stress in the fetus during labour. Measurements of the F2-isoprostane concentrations in cord blood at labour provide a clinically useful method to assess the perinatal outcome.

Acid-Base Imbalance↗

Unexpected loss of bipolar pacing with implanted dual chamber pacemakers.

Bipolar leads are most commonly used in the current practice of pacemaker therapy. In our study of 124 patients implanted with Guidant/Cardiac Pacemakers (CPI) Vigor dual chamber pacemakers, 5 patients had unexpectedly abrupt increases in bipolar lead impedance and pacing threshold 2 weeks to 18 months postimplantation without changes in sensing function. With the lead configuration reprogrammed to unipolar, the lead impedance and pacing threshold were restored to appropriate ranges. The changes in bipolar lead parameters can be caused by the CPI's "Quick Connect" (QC1) header lead system incorporated in these pacemakers.

Aged↗

Functional tissue engineering of articular cartilage through dynamic loading of chondrocyte-seeded agarose gels.

Due to its avascular nature, articular cartilage exhibits a very limited capacity to regenerate and to repair. Although much of the tissue-engineered cartilage in existence has been successful in mimicking the morphological and biochemical appearance of hyaline cartilage, it is generally mechanically inferior to the natural tissue. In this study, we tested the hypothesis that the application of dynamic deformational loading at physiological strain levels enhances chondrocyte matrix elaboration in cell-seeded agarose scaffolds to produce a more functional engineered tissue construct than in free swelling controls. A custom-designed bioreactor was used to load cell-seeded agarose disks dynamically in unconfined compression with a peak-to-peak compressive strain amplitude of 10 percent, at a frequency of 1 Hz, 3 x (1 hour on, 1 hour off)/day, 5 days/week for 4 weeks. Results demonstrated that dynamically loaded disks yielded a sixfold increase in the equilibrium aggregate modulus over free swelling controls after 28 days of loading (100 +/- 16 kPa versus 15 +/- 8 kPa, p < 0.0001). This represented a 21-fold increase over the equilibrium modulus of day 0 (4.8 +/- 2.3 kPa). Sulfated glycosaminoglycan content and hydroxyproline content was also found to be greater in dynamically loaded disks compared to free swelling controls at day 21 (p < 0.0001 and p = 0.002, respectively).

Animals↗

P2Y(2) receptor-stimulated phosphoinositide hydrolysis and Ca(2+) mobilization in tracheal epithelial cells.

Extracellular nucleotides have been implicated in the regulation of secretory function through the activation of P2 receptors in the epithelial tissues, including tracheal epithelial cells (TECs). In this study, experiments were conducted to characterize the P2 receptor subtype on canine TECs responsible for stimulating inositol phosphate (InsP(x)) accumulation and Ca(2+) mobilization using a range of nucleotides. The nucleotides ATP and UTP caused a concentration-dependent increase in [(3)H]InsP(x) accumulation and Ca(2+) mobilization with comparable kinetics and similar potency. The selective agonists for P1, P2X, and P2Y(1) receptors, N(6)-cyclopentyladenosine and AMP, alpha,beta-methylene-ATP and beta, gamma-methylene-ATP, and 2-methylthio-ATP, respectively, had little effect on these responses. Stimulation of TECs with maximally effective concentrations of ATP and UTP showed no additive effect on [(3)H]InsP(x) accumulation. The response of a maximally effective concentration of either ATP or UTP was additive to the response evoked by bradykinin. Furthermore, ATP and UTP induced a cross-desensitization in [(3)H]InsP(x) accumulation and Ca(2+) mobilization. These results suggest that ATP and UTP directly stimulate phospholipase C-mediated [(3)H]InsP(x) accumulation and Ca(2+) mobilization in canine TECs. P2Y(2) receptors may be predominantly mediating [(3)H]InsP(x) accumulation, and, subsequently, inositol 1,4,5-trisphosphate-induced Ca(2+) mobilization may function as the transducing mechanism for ATP-modulated secretory function of tracheal epithelium.

Adenosine Diphosphate↗

Linking African American mothers across life stage and station through photovoice.

Homeless young African American women and low-income, elderly African American women have housing needs specific to their age cohorts, yet they also have parallel and complementary housing, health, and personal needs. The young struggle to afford decent housing, while the old may have difficulty maintaining their homes. In this pilot study, intergenerational contact was established between young homeless women and elderly independently housed women through photovoice. Over six months, five African American women discussed photographs they had taken that focused on their current living arrangements and activities. Although the women spanned three generations, had different life experiences, and resided in a variety of home settings, the sharing of photographs revealed many commonalities. In the process of discussing photographs, the women established mutual respect, exercised reciprocal affirmation, and built alliances. The preliminary study is preparatory to exploring the feasibility of establishing house-sharing arrangements for mutual assistance between these cohorts.

Adult↗

Laparosopic removal of large gastric phytobezoars.

Large gastric bezoars are difficult to remove endoscopically. A 78-year-old man presenting with abdominal pain and loss of appetite for 4 months was admitted and evaluated. Gastroscopy disclosed two large phytobezoars within the stomach. Laparoscopic removal was undertaken. The bezoars were removed via a gastrotomy using the three-trocar technique. They were successfully retrieved from the abdominal cavity using an improvised "endobag" made from a simple surgical glove. Such an endobag presents several advantages; they are easy to make, sterile, economical, readily available, disposable, there is ample space to manipulate the specimen within, and there is minimal risk of contamination throughout the procedure. The authors recommend this approach for the treatment of patients with large gastric bezoars in whom laparotomy is indicated.

Abdominal Pain↗

Comparison of clinical behaviors and responses to radiation between squamous cell carcinomas and adenocarcinomas/adenosquamous carcinomas of the cervix.

BACKGROUND: Adenocarcinomas/adenosquamous carcinomas (AC/ASC) are relatively uncommon histological subtypes in cervical cancer. In this study, we retrospectively compared the clinical behaviors and responses to radiation of squamous cell carcinomas (SCC) and AC/ASC in patients with stage I-IVA cervical cancer primarily treated by radiotherapy (RT). METHODS: Nine hundred twenty-eight patients with cervical cancer primarily treated with RT were included in this study. Ninety-four percent of the patients had SCC and 6% had AC/ASC. The association of histological subtype to various clinical parameters was assessed. Tumor response and failure patterns were analyzed, and the prognostic significance of histological subtype and other clinical parameters were evaluated by univariate and multivariate analyses. RESULTS: (1) Patients with AC/ASC had a significantly higher percentage of bulky I-IIA disease, a younger age (< 45 years), and an elevated carcinoembryonic antigen (CEA) level, but a lower percentage of elevated squamous cell carcinoma antigen (SCC-Ag) level, than patients with SCC. Hemoglobin (Hb) levels and lymph node status were not significantly different between SCC and AC/ASC patients. (2) A higher percentage of AS/ASC patients had residual induration 2 to 3 months after RT than those with SCC (40% vs. 21.6%, p = 0.008). Higher local failure rates were found for AC/ASC patients compared to SCC patients (38% vs. 15%, p = 0.001). (3) The 5-year disease-specific survival rate was 66% and 50% for SCC and AC/ASC patients, respectively (p = 0.016). Advanced stages, Hb levels lower than 10 g/dl, positive lymph nodes, AC/ASC histological subtype, and SCC-Ag levels higher than 10 ng/ml were independent prognostic factors for worse survival for all patients. For patients with AC/ASC in particular, only stage and positive lymph nodes were significant prognostic factors. CONCLUSION: Cervical cancer patients with AC/ASC are relatively younger and have a higher incidence of elevated CEA and a lower incidence of elevated SCC-Ag levels than those with SCC. For AC/ASC patients, the tumor responses to RT were relatively slow and poor. This group of patients had a worse survival rate than patients with SCC, mainly because of the higher incidence of uncontrolled local disease for AC/ASC patients.

Adenocarcinoma↗

Successful management of massive blood loss to extremely low hemoglobin in an elderly woman receiving spinal surgery.

Blood transfusion is absolutely indicated in acute anemia when the hemoglobin concentration falls below 6 g/dL. Sometimes it challenges the anesthesiologists if the blood intended for urgent transfusion is not readily or quickly available. In this case report, we describe an 81-year-old lady who accidentally sustained acute anemia after spinal surgery with the hemoglobin concentration falling to 1.4 g/dL. During the long wait for the process of cross-matching tests and delivery of blood from the blood bank in the city remote from the hospital, we could do nothing but administer crystalloid and colloid solutions to maintain the circulatory volume to prevent low cardiac output. Epinephrine was given when systolic blood pressure fell below 70 mmHg. Central venous pressure and arterial blood pressure were monitored to guide all the treatment. Fortunately, patient fully recovered on postoperative day 3 without any adverse events.

Aged↗

Primary staphylococcal infection and toxic shock syndrome diagnosed by polymerase chain reaction.

Primary staphylococcal pneumonia complicated with toxic shock syndrome (TSS) is relatively uncommon in children. Staphylococcus aureus exotoxins are thought to function as superantigens, and seem to promote disease manifestations. The identification of staphylococcal toxin genes by polymerase chain reaction (PCR) offers a specific and rapid diagnostic method for TSS. We describe a 7-year-old child with TSS resulting from staphylococcal pneumonia. S. aureus enterotoxins A and B were detected in the sputum of this patient by PCR.

Child↗

Post-traumatic, high-flow priapism treated with selective cavernous artery embolization and intracavernous streptokinase irrigation: a case report.

A 54-year-old man developed priapism shortly after a blunt perineal trauma. An arteriocavernous fistula caused the high-flow priapism, and was detected on both color Doppler sonography and selective phaloarteriography. Selective embolization of the left cavernous artery with Gelfoam was performed to seal the fistula, resulting in immediate detumescence. However, the penis remained firm despite returning to almost normal size. No fistula was detected by subsequent color Doppler sonography and phaloarteriography examination. Intracavernous irrigation with 200,000 U streptokinase was applied to treat residual firmness 2 weeks after embolization. Successful sexual intercourse was reported 3 months later. The combination of selective cavernous artery embolization and intracavernous streptokinase irrigation was effective for the treatment of the high-flow priapism in this case.

Arteries↗

Ascending projections from the area around the spinal cord central canal: A Phaseolus vulgaris leucoagglutinin study in rats.

A single small iontophoretic injection of Phaseolus vulgaris leucoagglutinin labels projections from the area surrounding the spinal cord central canal at midthoracic (T6-T9) or lumbosacral (L6-S1) segments of the spinal cord. The projections from the midthoracic or lumbosacral level of the medial spinal cord are found: 1) ascending ipsilaterally in the dorsal column near the dorsal intermediate septum or the midline of the gracile fasciculus, respectively; 2) terminating primarily in the dorsal, lateral rim of the gracile nucleus and the medial rim of the cuneate nucleus or the dorsomedial rim of the gracile nucleus, respectively; and 3) ascending bilaterally with slight contralateral predominance in the ventrolateral quadrant of the spinal cord and terminating in the ventral and medial medullary reticular formation. Other less dense projections are to the pons, midbrain, thalamus, hypothalamus, and other forebrain structures. Projections arising from the lumbosacral level are also found in Barrington's nucleus. The results of the present study support previous retrograde tract tracing and physiological studies from our group demonstrating that the neurons in the area adjacent to the central canal of the midthoracic or lumbosacral level of the spinal cord send long ascending projections to the dorsal column nucleus that are important in the transmission of second-order afferent information for visceral nociception. Thus, the axonal projections through both the dorsal and the ventrolateral white matter from the CC region terminate in many regions of the brain providing spinal input for sensory integration, autonomic regulation, motor and emotional responses, and limbic activation.

Animals↗

Metal regulation of metallothionein participation in redox reactions.

Like glutathione or dithiothreitol, metallothionein effects the formation of pancreatic ribonuclease A from its S-sulfonated derivative catalyzed by protein disulfide isomerase. EDTA increases the yield of ribonuclease A activity recovery with metallothionein but does not affect the reaction with glutathione or dithiothreitol. EDTA also increases the reactivity of thiol groups in metallothionein with 5,5'-dithiobis-(2-nitrobenzoic acid) by chelation of zinc ions. It is suggested that the thiol groups in metallothionein form a part of the pool of cellular thiols in the regulation of cellular redox reactions and their availability is modulated by zinc chelation.

Animals↗

alpha5 subunit in Trypanosoma brucei proteasome can self-assemble to form a cylinder of four stacked heptamer rings.

The proteasomes have a central role in catalysing protein degradation among both prokaryotes and eukaryotes. The 20 S proteasome constitutes their catalytic core. In studying the structure of Trypanosoma brucei 20 S proteasomes, we isolated by two-dimensional (2D) gel electrophoresis a 27 kDa subunit protein with an estimated pI of 4.7 and subjected it to mass spectrometric analysis. A tryptic peptide sequence from the protein was found identical with that of the rat alpha5 subunit. With the use of antiserum against T. brucei 20 S proteasomes to screen a T. b. rhodesiense lambda expression cDNA library, we obtained a cDNA clone encoding a full-length protein of 246 amino acid residues with a calculated molecular mass of 27174 Da and a pI of 4.71. It bears 50. 0% and 46.3% sequence identity with rat and yeast proteasome subunit alpha5 respectively, and matches all the peptide sequences derived from MS of the 2D gel-purified protein. The protein is thus designated the alpha5 subunit of T. brucei 20 S proteasome (TbPSA5). The recombinant protein, expressed in plasmid-transformed Escherichia coli, was found in a 27 kDa monomer form as well as polymerized forms with estimated molecular masses ranging from 190 to 800 kDa. Under the electron microscope, the most highly polymerized forms bear the appearance of cylinders of four-stacked heptamer rings with an estimated outer diameter of 14.5 nm and a length of 18 nm, which were immunoprecipitable by anti-(T. brucei 20 S proteasome) antiserum. In view of the documented self-assembly of the archaeon proteasome alpha subunit into double heptamer rings and the spontaneous assembly of the two alpha subunits from the 20 S proteasome of Rhodococcus erythropolis, the self-assembly of the T. brucei alpha subunit might reflect a common feature of proteasome biogenesis shared by prokaryotes and primitive eukaryotes such as the trypanosomes but apparently lost among the higher forms of eukaryote such as the yeast and the mammals.

Amino Acid Sequence↗

Structural and functional characterizations of the proteasome-activating protein PA26 from Trypanosoma brucei.

The activated 20 S proteasome, which has been found only in mammalian cells, is composed of two heptamer rings of an activator protein on each end of the 20 S proteasome and is inducible by interferon-gamma. A 20 S proteasome has been recently identified in a protozoan pathogen Trypanosoma brucei, but there has been no experimental evidence yet for the presence of a 26 S proteasome. Instead, an activated form of 20 S proteasome was isolated from this organism, which has significantly enhanced peptidase activities. It consists of an additional activator protein with an estimated molecular mass of 26 kDa (PA26) (To, W. Y., and Wang, C. C. (1997) FEBS Lett. 404, 253-262). The profile and sequences of tryptic peptides from PA26 were determined by mass spectrometry; no matches were found in the data base. The peptide sequences were used in reverse transcriptase-polymerase chain reaction to isolate a full-length cDNA clone encoding PA26. The protein sequence thus derived from it indicates little sequence identity with those of mammalian activator proteins PA28 alpha, beta, or gamma. There is only a single copy of PA26 gene in T. brucei. Purified recombinant PA26 polymerizes spontaneously to form heptamer ring with an outer diameter of 8.5 nm. The ring binds and activates 20 S proteasomes from T. brucei as well as rat, whereas human PA28alpha can neither bind nor activate T. brucei 20 S proteasome. The former is thus apparently more ubiquitous than PA28 in its capability of binding to and activating 20 S proteasomes. Its presence in T. brucei may also suggest a more ancient origin of proteasome activator proteins and a much wider involvement in protein degradation among other eukaryotic organisms than was originally envisaged.

Amino Acid Sequence↗