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Biomedical subjects

C Park

Publications and source records attributed to C Park.

At least 19 recordsLinked to original sources

Protein stitchery: design of a protein for selective binding to a specific DNA sequence.

We present a general strategy for designing proteins to recognize DNA sequences and illustrate this with an example based on the "Y-shaped scissors grip" model for leucine-zipper gene-regulatory proteins. The designed protein is formed from two copies, in tandem, of the basic (DNA binding) region of v-Jun. These copies are coupled through a tripeptide to yield a "dimer" expected to recognize the sequence TCATCGATGA (the v-Jun-v-Jun homodimer recognizes ATGACTCAT). We synthesized the protein and oligonucleotides containing the proposed binding sites and used gel-retardation assays and DNase I footprinting to establish that the dimer binds specifically to the DNA sequence TCATCGATGA but does not bind to the wild-type DNA sequences, nor to oligonucleotides in which the recognition half-site is modified by single-base changes. These results also provide strong support for the Y-shaped scissors grip model for binding of leucine-zipper proteins.

Amino Acid Sequence

Treatment of postcataract fibrinous membranes with tissue plasminogen activator.

PURPOSE: The purpose of this study is to investigate the efficacy and toxicity of tissue plasminogen activator (tPA) in treatment of postcataract fibrinous membranes. METHODS: The authors injected 25 micrograms of tPA into the anterior chamber of 52 pseudophakic eyes with moderate to severe fibrinous membranes that developed after cataract surgery. RESULTS: Intraocular tPA injection resulted in complete fibrinolysis in 47 of 52 eyes (90%) and partial fibrinolysis in 5 of 52 eyes (10%). In cases of complete fibrinolysis, resolution of the fibrinous membrane occurred within 30 minutes in 22 of 47 eyes (47%) and within 1 hour in 25 of 47 eyes (53%). No evidence of toxicity was observed as measured by slit-lamp biomicroscopy, intraocular pressure, and corneal endothelial cell density, size, and morphology. Complications of tPA administration included periorbital pain (4 eyes), anterior chamber hemorrhage (4 eyes), and anterior chamber turbidity (3 eyes). CONCLUSION: The authors believe that intraocular tPA is effective and safe in the treatment of postcataract fibrinous membranes.

Adult

Arterial supply of the anterior ear.

Twenty cadaver auricles were injected with a latex solution to define the arterial supply of the anteroauricular surface. Two arterial networks exist, the network of the triangular fossa-scapha and the network of the concha. Both eventually communicate on the anthelix. The triangular fossa-scapha network originates from one subbranch of the upper auricular branch of the superficial temporal artery and from branches of the posterior auricular artery that come through the earlobe and triangular fossa and over the helical margin. The conchal network is provided by two to four perforators that come from the posterior auricular artery, piercing the conchal floor. Auricular branches of the superficial temporal artery in the preauricular region and their communications with the posterior auricular artery also were confirmed. We believe that a greater understanding of the detailed arterial anatomy in this area allows one to develop safely a variety of surgical techniques for reconstruction of the ear.

Arteries

Involvement of SecB, a chaperone, in the export of ribose-binding protein.

Ribose-binding protein (RBP) is an exported protein of Escherichia coli that functions in the periplasm. The export of RBP involves the secretion machinery of the cell, consisting of a cytoplasmic protein, SecA, and the integral membrane translocation complex, including SecE and SecY. SecB protein, a chaperone known to mediate the export of some periplasmic and outer membrane proteins, was previously reported not to be involved in RBP translocation even though small amounts of in vitro complexes between SecB and RBP have been detected. In our investigation, it was shown that a dependence on SecB could be demonstrated under conditions in which export was compromised. Species of RBP which carry two mutations, one in the leader that blocks export and a second in the mature protein which partially suppresses the export defect, were shown to be affected by SecB for efficient translocation. Five different changes which suppress the effect of the signal sequence mutation -17LP are all located in the N domain of the tertiary structure of RBP. All species of RBP show similar interaction with SecB. Furthermore, a leaky mutation, -14AE, generated by site-specific mutagenesis causes reduced export in the absence of SecB. These results indicate that SecB can interact with RBP during secretion, although it is not absolutely required under normal circumstances.

Bacterial Proteins

Subacute necrotizing lymphadenitis--a collective clinicopathological and immunohistochemical study.

Subacute necrotizing lymphadenitis (SNL) is a well documented and unique clinicopathologic entity, although its etiology and pathogenesis have not been clearly established. Microscopically, cortical and paracortical necrotizing lesions with karyorrhexis, abundant nuclear debris and infiltration of large mononuclear cells are characteristic. This study analyzed the common clinical and pathological features of 118 patients with SNL and the nature of the mononuclear cells. Patients were generally young women and revealed cervical lymphadenopathy with tenderness, fever, leukopenia and elevation of the erythrocyte sedimentation rate. Features of the adjacent uninvolved area in the lymph node included a starry sky pattern, follicle centers, sinus histiocytosis or aggregation of foamy histicoytes. There was an inverse relationship between the extent of necrosis and of histocytic infiltration but not between the extent of necrosis and the duration from the onset of symptoms to the diagnosis. Immunohistochemically the infiltrated mononuclear cells of the affected foci were T lymphocytes and histiocytes. The clinical, histological and immunohistochemical features suggest that SNL represents a hypersensitivity reaction to certain infectious agent without forming granuloma.

Adolescent

DNA flow cytometry in pheochromocytoma and paraganglioma.

Flow cytometric DNA analysis was performed on 19 adrenal pheochromocytomas and 6 extra-adrenal paragangliomas in parallel with clinical and histopathological review to determine the usefulness of this technique to predict biologic behavior of these tumors. In pheochromocytomas and paragangliomas, tetraploidy or near-tetraploidy occurred in 32% and 33% and aneuploidy in 10% and none respectively. A case of malignant pheochromocytoma had diploid DNA content. Occurrence of aneuploidy or tetraploidy is frequent in clinically benign tumors in conjunction with a marked degree of nuclear atypia and cannot be a predictor of malignancy.

Adolescent

Mutations that affect the folding of ribose-binding protein selected as suppressors of a defect in export in Escherichia coli.

It has been proposed (Randall, L. L., and Hardy, S. J. S. (1986) Cell 46, 921-928) that export of protein involves a kinetic partitioning between the pathway that leads to productive export and the pathway that leads to the folding of polypeptides into a stable conformation that is incompatible with export. As predicted from this model, a decrease in the rate of export of maltose-binding protein to the periplasmic space in Escherichia coli resulting from a defect in the leader sequence was able to be partially overcome by a mutation that slowed the folding of the precursor, thereby increasing the time in which the polypeptide was competent for export. (Liu, G., Topping, T. B., Cover, W. H., and Randall, L. L. (1988) J. Biol. Chem. 263, 14790-14793). Here we describe mutations of the gene encoding ribose-binding protein that were selected as suppressors of a defect in export of that protein and that alter the folding pathway. We propose that selection of such suppressors may provide a general method to obtain mutations that affect the folding properties of any protein that can be expressed and exported in E. coli.

Biological Transport

Contaminant reduction by ventilation in a confined space model--toxic concentrations versus oxygen deficiency.

Airborne contaminants can create hazardous conditions in confined spaces (CS) across a broad range of concentrations, e.g., from relatively low, potentially toxic levels (ppm) to much higher levels (%) causing oxygen deficiency. This study investigated ventilation characteristics for isobutylene (IBE) at relatively low concentrations, simulating toxic levels. Experimental data were compared to results from previous studies of oxygen deficiency. Data were obtained at several locations in a cubical CS model, with several variable test parameters: ventilation mode (exhaust and supply), volume flow rate ("air changes" per hour), and ventilation inlet/outlet elevation (% of model height). Findings indicated similar ventilation characteristics, in general, for simulated toxic (IBE) levels compared to oxygen deficiency. Both IBE and O2 deficiency data have shown that supply ventilation is typically more effective than exhaust and that CS locations aligned with supply outlets experience much more rapid contaminant reduction than do other locations. The data suggest that highly accurate predictions of ventilation characteristics cannot be expected for all cases with widely different contaminants and concentrations. Findings from this study indicate that ventilation guidelines for one range of contaminant concentration (e.g., causing oxygen deficiency) can be extended reasonably to encompass a broader range of concentration (e.g., to include toxic or flammable atmospheres).

Air Pollutants, Occupational

Total palmar resurfacing with scapular free flap in a 26-year contracted hand.

The scapular free flap offers a large-sized and well-vascularized coverage for variable defects. It is not regarded, however, as a favorite tool for palmar resurfacing because of its bulky and rigid nature. A 28-year-old man had sustained a contact dermal injury at the age of 2 years that resulted in a severely deformed hand. The total palmar defect was resurfaced by surgical intervention with a scapular free flap. After secondary procedures, the final result was promising. The present report focuses on the secondary procedures, which make a more functional and aesthetically pleasing hand after coverage with the scapular flap. Also, our result suggests that even finger joints contracted for 26 years can recover motion if they have not been directly damaged.

Adult

Functioning free latissimus dorsi muscle transplantation: anterogradely positioned usage in reconstruction of extensive forearm defect.

We present a patient with successful functional recovery of a severely injured upper extremity after a microneurovascular free latissimus dorsi muscle and combined composite skin were transplanted with an anterograde position. In our patient, the anterogradely positioned usage of the lastissimus muscle offered two major advantages; one advantage was the direct anastomosis of a more proximal healthy uninjured recipient nerve without interpositional nerve graft, and the other, one-stage coverage including the elbow wound with combined scapular skin flap. From our result, we deduce that a neurovascular free latissimus dorsi muscle transplantation with an anterograde position is selectively indicated in functional recovery of an extensively injured forearm.

Adult

A single-stage two-flap method of total ear reconstruction.

A single-stage two-flap method of total ear reconstruction in congenital microtia is reported. This method was derived from the one-stage reconstruction described by Song and Song. Two flaps defined by vascular basis were elevated on the mastoid area: the superficial skin flap supplied mainly by subcutaneous pedicled arteriole perforators from the posterior auricular artery and the deeper axial-pattern fascial flap including the posterior auricular artery itself. The ear framework, exaggeratedly carved using autologous rib cartilage, could be inserted easily between the two flaps, simultaneously producing the auriculocephalic angle and the conchal wall. Intraoperative expansion of the skin flap and postoperative external ear molding also were performed to create aesthetically pleasing ears.

Adolescent

Evaluation of 99mTc-hexamethylpropyleneamine oxime cerebral blood flow mapping after acute focal ischemia in rats.

Although the blood flow tracer hexamethylpropyleneamine oxime is now widely used for cerebral blood flow mapping using single-photon emission computed tomography, its uptake into acutely ischemic brain has not been well studied. We performed a double-label autoradiographic study in which 99mTc-hexamethylpropyleneamine oxime uptake was compared with [14C]iodoantipyrine-derived cerebral blood flow 0.5, 24, and 72 hours after middle cerebral artery occlusion in 11 rats. We noted excellent correspondence between iodoantipyrine and hexamethylpropyleneamine oxime autoradiograms at all times with both simultaneous tracer injection and 30-minute separation of tracer injections. When the ischemic and hyperemic areas were measured from the same brain section using the two different tracers, hexamethylpropyleneamine oxime underestimated the iodoantipyrine-derived areas by less than 1% (95% confidence interval -2.9% to 2.3%). The maximum discrepancy (-19%) was seen at high flows. When the two tracers were injected separately, the uptake of hexamethylpropyleneamine oxime was not linear compared with iodoantipyrine-derived cerebral blood flow, but the relation became linear after the Lassen correction factor was applied. Hexamethylpropyleneamine oxime uptake thus accurately represents cerebral blood flow 0.5-72 hours after acute cerebral ischemia.

Animals

Normal splenic size in infants and children: sonographic measurements.

The purpose of this study was to establish guidelines for normal splenic size at different ages by using a simple and reproducible sonographic method. Two hundred thirty patients, from neonate to 20-year-old, had sonography because of abdominal and/or pelvic problems unrelated to the spleen. Findings on sonograms of the liver and kidneys were normal in all cases. Splenic size was measured by obtaining a coronal view that included the hilum, while the patient was breathing quietly. The greatest longitudinal distance between the dome of the spleen and the tip (splenic length) was measured and correlated with age, height, and weight. The following guidelines are proposed for the upper limit of normal splenic length based on this simple, easy to use, one-measurement technique: splenic length no greater than 6.0 cm at 3 months, 6.5 cm at 6 months, and 7.0 cm at 12 months, 8.0 cm at 2 years, 9.0 at 4 years, 9.5 cm at 6 years, 10.0 cm at 8 years, 11.0 cm at 10 years, 11.5 cm at 12 years, 12.0 cm at 15 years or older for girls, and 13.0 cm at 15 years or older for boys. Twenty-two patients with known abnormalities of the spleen were randomly selected and their splenic lengths compared with the proposed guidelines; in each case, the length of the spleen exceeded the upper limit of normal for that age. Normal values of a single measurement of the greatest longitudinal diameter of the spleen, from the dome to the tip measured at the hilum in the coronal plane, were obtained in patients from newborn to 20 years old.

Adolescent

Effect of a modified rectus tuck on anterior ciliary artery perfusion.

Strabismus surgery results in the permanent interruption of anterior ciliary blood flow, predisposing the eye to anterior segment ischemia (ASI). A primate model was used to assess the effectiveness of a new muscle-scleral tuck for preserving anterior ciliary artery circulation. The model consisted of removing 3 rectus muscles from both eyes of 2 rhesus monkeys, then performing a tuck on the inferior rectus (IR) right eye while leaving the left IR as a control. Four weeks later, a modified tuck was performed on the virgin left IR. Fluorescein iris angiograms of both eyes were obtained, and preoperative angiograms at 5-15 sec. showed normal 360 degrees perfusion. Postoperative follow-up angiograms showed segmental superior temporal filling defects and preservation of perfusion in the distribution of the IR. Comparison of fellow eyes tucked vs control IR showed no difference in the filling pattern in both monkeys. Comparison of the same eye before and after tuck also showed essentially the same filling pattern in all 4 eyes with preservation of inferior circulation. Our conclusion is that the modified tuck preserves the anterior ciliary blood flow and may be useful as a muscle-strengthening procedure in patients predisposed to developing ASI.

Animals

Effects of iodinated fatty acid ester on human hepatocellular carcinoma cells.

The interaction between Lipiodol and cells was studied by treating Lipiodol in a human hepatocellular carcinoma cell line(Hep) and mouse fibroblast cell line (L929). Irregular, sustained radioactivity was released from both cell lines shortly after incubation in the radioiodinated Lipiodol mixed media. Lipiodol droplets were found to be firmly attached to the cells following the incubation and these cells were strongly positive for fat stains. The radioiodinated Lipiodol demonstrated the same behavior of accumulation within the cell and on the cell membrane. Although the amount of Lipiodol attached was almost equal in both of the cell lines, the final amount accumulated in the cells was larger in the Hep cells. The accumulation of Lipiodol within the cell and on the cell membrane may play a significant role for its selective targeting and its prolonged retention in the solid tumor.

Carcinoma, Hepatocellular