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Biomedical subjects

M G Lee

Publications and source records attributed to M G Lee.

At least 91 records · Page 5Linked to original sources
PubMed1999

Suppression of rat hepatic cytochrome P450s by protein-calorie malnutrition: complete or partial restoration by cysteine or methionine supplementation.

Pharmacokinetic profiles of therapeutic agents are altered by protein-calorie malnutrition (PCM). The current study was designed to determine the expression of hepatic cytochrome P450s in rats after protein restriction and to investigate its molecular basis. Western blot analysis revealed that rats with protein restriction for 4 weeks exhibited marked suppression in the hepatic P450 1A2, 2C11, 2E1, and 3A1/2 levels. Northern blot analysis showed that hepatic P450 1A2, 2C11, and 3A1/2 mRNAs were significantly decreased in the state of PCM. The P450 2E1 mRNA level was slightly decreased in PCM rats, suggesting the possibility that expression of P450 2E1 affected by PCM might result from the transcriptional and/or posttranscriptional regulation. PCM-induced changes in most P450 expression completely or partially returned to control levels by a week of cysteine supplementation. Cysteine also prevented decreases in P450 1A2, 2C11, 2E1, and 3A1/2 mRNA levels by PCM. Methionine was minimally active in restoring the P450 expression. A metabolic change in hepatic ethoxyresorufin dealkylase activity in PCM rats was consistent with the P450 apoprotein and mRNA levels. Although the plasma concentrations of azosemide, a loop diuretic, primarily metabolized by cytochrome P450 1A, increased in protein-deprived rats, cysteine supplementation significantly reduced the increased plasma concentrations of the drug. The altered pharmacokinetic parameters of azosemide in PCM rats returned to those of control after cysteine supplementation, corroborating the conclusion that cysteine was effective in restoring cytochrome P450 expression and metabolic activities.

Animals↗
PubMed1999

Insights into eukaryotic multistep phosphorelay signal transduction revealed by the crystal structure of Ypd1p from Saccharomyces cerevisiae.

"Two-component" phosphorelay signal transduction systems constitute a potential target for antibacterial and antifungal agents, since they are found exclusively in prokaryotes and lower eukaryotes (yeast, fungi, slime mold, and plants) but not in mammalian organisms. Saccharomyces cerevisiae Ypd1p, a key intermediate in the osmosensing multistep phosphorelay signal transduction, catalyzes the phosphoryl group transfer between response regulators. Its 1.8 A structure, representing the first example of a eukaryotic phosphorelay protein, contains a four-helix bundle as in the HPt domain of Escherichia coli ArcB sensor kinase. However, Ypd1p has a 44-residue insertion between the last two helices of the helix bundle. The side-chain of His64, the site of phosphorylation, protrudes into the solvent. The structural resemblance between Ypd1p and ArcB HPt domain suggests that both prokaryotes and lower eukaryotes utilize the same basic protein fold for phosphorelay signal transduction. This study sheds light on the best characterized eukaryotic phosphorelay system.

Amino Acid Sequence↗
PubMed1999

The procyclin repertoire of Trypanosoma brucei. Identification and structural characterization of the Glu-Pro-rich polypeptides.

The surface of the insect stages of the protozoan parasite Trypanosoma brucei is covered by abundant glycosyl phosphatidylinositol (GPI)-anchored glycoproteins known as procyclins. One type of procyclin, the EP isoform, is predicted to have 22-30 Glu-Pro (EP) repeats in its C-terminal domain and is encoded by multiple genes. Because of the similarity of the EP isoform sequences and the heterogeneity of their GPI anchors, it has been impossible to separate and characterize these polypeptides by standard protein fractionation techniques. To facilitate their structural and functional characterization, we used a combination of matrix-assisted laser desorption ionization and electrospray mass spectrometry to analyze the entire procyclin repertoire expressed on the trypanosome cell. This analysis, which required removal of the GPI anchors by aqueous hydrofluoric acid treatment and cleavage at aspartate-proline bonds by mild acid hydrolysis, provided precise information about the glycosylation state and the number of Glu-Pro repeats in these proteins. Using this methodology we detected in a T. brucei clone the glycosylated products of the EP3 gene and two different products of the EP1 gene (EP1-1 and EP1-2). Furthermore, only low amounts of the nonglycosylated products of the GPEET and EP2 genes were detected. Because all procyclin genes are transcribed polycistronically, the latter finding indicates that the expression of the GPEET and EP2 genes is post-transcriptionaly regulated. This is the first time that the whole procyclin repertoire from procyclic trypanosomes has been characterized at the protein level.

Amino Acid Sequence↗
PubMed1999

Effects of satratoxins and other macrocyclic trichothecenes on IL-2 production and viability of EL-4 thymoma cells.

The macrocyclic trichothecenes are a group of potent protein synthesis inhibitors that have been encountered in indoor air and food as a result of infestation by the fungus Stachybotrys. To evaluate the capacity of these mycotoxins to alter immune functions, the effects of satratoxin G, H, F, roridin A, and verrucarin A on interleukin 2 (IL-2) production and viability were evaluated in a murine T-cell model. EL-4 thymoma cells were stimulated with phorbol 12-myristate 13-acetate and ionomycin and concurrently exposed to various concentrations of the trichothecenes. Enzyme-linked immunosorbent assay (ELISA) of supernatants revealed that IL-2 concentrations at 24 and 72 h were significantly increased in cultures that were incubated in the presence of 0.5 to 1 ng/ml of satratoxin H, 1 to 5 ng/ml of isosatratoxin F, 0.1 to 0.5 ng/ml of roridin A, and 0.25 to 0.5 ng/ml of verrucarin A. However, IL-2 levels at these time points were significantly depressed when incubated in the presence of higher concentrations of satratoxin G (> or =2.5 ng/ml), satratoxin H and isosatratoxin F (> or =5 ng/ml), and roridin A and verrucarin A (> or =1 ng/ml). Cell viability, as measured by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay, was depressed by each of the trichothecenes in a concentration-dependent manner. MTT responses were significantly decreased by as little as 0.5 ng/ml satratoxin G, roridin A, and verrucarin A and by 2.5 ng/ml of isosatratoxin F and satratoxin H. When these data were compared to those found in EL-4 cells for the 8-ketotrichothecene vomitoxin (deoxynivalenol), a common food contaminant, the macrocyclic trichothecenes were at least 100 times more potent. The results indicate that, at low concentrations, macrocyclic trichothecenes as a group could superinduce IL-2 production even while partially decreasing cell viability, whereas higher concentrations suppressed cytokine production and were markedly cytotoxic. The capacity of these compounds to dysregulate cytokine production in a biphasic fashion may play an etiologic role in outbreaks of human illnesses associated with indoor Stachybotrys contamination.

Air Microbiology↗
PubMed1999

Effect of a new chemoprotective agent, 2-(allylthio)pyrazine, on the pharmacokinetics of intravenous theophylline in rats.

The effect of 2-(allylthio)pyrazine (2-AP) pretreatment on the pharmacokinetics of theophylline and its metabolites was investigated after 1-min intravenous administration of aminophylline, 5 mg/kg as theophylline, to rats pretreated with three consecutive daily oral administration of 2-AP, 100 mg/kg. The AUC0-2 h of a metabolite of theophylline, 1,3-dimethyluric acid, 1,3-DMU (62.3 versus 106 microg/min per ml), and the percentages of intravenous dose of theophylline excreted in 24-h urine as 1,3-DMU (12.4% versus 20.8%, expressed in terms of theophylline) decreased significantly in 2-AP-pretreated rats when compared with those in control rats. Since CYP1A2 and CYP2E1 are involved in the formation of 1,3-DMU from theophylline, and 2-AP considerably suppressed CYP2E1 and tended to suppress CYP1A2 in rats, decreased formation of 1,3-DMU in 2-AP-pretreated rats could be mainly due to suppression of the CYP2E1 expression by pretreatment with 2-AP.

Animals↗
PubMed1999

Acquisition of lipoproteins in the procyclic form of Trypanosoma brucei.

The procyclic form of Trypanosoma brucei binds and internalizes bovine high density lipoprotein (HDL) particles in a saturable process; the binding and uptake of (125)I-labeled HDL are inhibited by excess unlabeled HDL. We calculated that each procyclic trypanosome exposes approximately 1.0 x 10(6) binding sites for bovine HDL, with an equilibrium dissociation constant (Kd) of approximately 1.26 x 10(-7) M. Uptake of HDL particles does not occur at 4 degrees C. At 28 degrees C, a significant amount of the internalized HDL particles were efficiently degraded through a process that is sensitive to the presence of 50 microM chloroquine. These results suggested that the uptake of HDL particles in procyclic T. brucei may occur via receptor mediated endocytosis, leading to proteolytic degradation of the particles in an acidic and endocytic compartment.

Animals↗
PubMed1999

Cystic fibrosis transmembrane conductance regulator regulates luminal Cl-/HCO3- exchange in mouse submandibular and pancreatic ducts.

We have demonstrated previously the regulation of Cl-/HCO3- exchange activity by the cystic fibrosis transmembrane conductance regulator (CFTR) in model systems of cells stably or transiently transfected with CFTR (Lee, M. G., Wigley, W. C., Zeng, W., Noel, L. E., Marino, C. R., Thomas, P. J., and Muallem, S. (1999) J. Biol. Chem. 274, 3414-3421). In the present work we examine the significance of this regulation in cells naturally expressing CFTR. These include the human colonic T84 cell line and the mouse submandibular gland and pancreatic ducts, tissues that express high levels of CFTR in the luminal membrane. As in heterologous expression systems, stimulation of T84 cells with forskolin increased the Cl-/HCO3- exchange activity independently of CFTR Cl- channel activity. Freshly isolated submandibular gland ducts from wild type mice showed variable Cl-/HCO3- exchange activity. Measurement of [Cl-]i revealed that this was largely the result of variable steady-state [Cl-]i. Membrane depolarization with 5 mM Ba2+ or 100 mM K+ increased and stabilized [Cl-]i. Under depolarized conditions wild type and DeltaF/DeltaF mice had comparable basal Cl-/HCO3- exchange activity. Notably, stimulation with forskolin increased Cl-/HCO3- exchange activity in submandibular gland ducts from wild type but not DeltaF/DeltaF mice. Microperfusion of the main pancreatic duct showed Cl-/HCO3- exchange activity in both the basolateral and luminal membranes. Stimulation of ducts from wild type animals with forskolin had no effect on basolateral but markedly stimulated luminal Cl-/HCO3- exchange activity. By contrast, forskolin had no effect on either basolateral or luminal Cl-/HCO3- exchange activity of ducts from DeltaF/DeltaF animals. We conclude that CFTR regulates luminal Cl-/HCO3- exchange activity in CFTR-expressing cells, and we discuss the possible physiological significance of these findings regarding cystic fibrosis.

Animals↗
PubMed1999

Dynamic association of proteasomal machinery with the centrosome.

Although the number of pathologies known to arise from the inappropriate folding of proteins continues to grow, mechanisms underlying the recognition and ultimate disposition of misfolded polypeptides remain obscure. For example, how and where such substrates are identified and processed is unknown. We report here the identification of a specific subcellular structure in which, under basal conditions, the 20S proteasome, the PA700 and PA28 (700- and 180-kD proteasome activator complexes, respectively), ubiquitin, Hsp70 and Hsp90 (70- and 90-kD heat shock protein, respectively) concentrate in HEK 293 and HeLa cells. The structure is perinuclear, surrounded by endoplasmic reticulum, adjacent to the Golgi, and colocalizes with gamma-tubulin, an established centrosomal marker. Density gradient fractions containing purified centrosomes are enriched in proteasomal components and cell stress chaperones. The centrosome-associated structure enlarges in response to inhibition of proteasome activity and the level of misfolded proteins. For example, folding mutants of CFTR form large inclusions which arise from the centrosome upon inhibition of proteasome activity. At high levels of misfolded protein, the structure not only expands but also extensively recruits the cytosolic pools of ubiquitin, Hsp70, PA700, PA28, and the 20S proteasome. Thus, the centrosome may act as a scaffold, which concentrates and recruits the systems which act as censors and modulators of the balance between folding, aggregation, and degradation.

Adenosine Triphosphatases↗
PubMed1999

Regulation of Cl-/ HCO3- exchange by cystic fibrosis transmembrane conductance regulator expressed in NIH 3T3 and HEK 293 cells.

A central function of cystic fibrosis transmembrane conductance regulator (CFTR)-expressing tissues is the secretion of fluid containing 100-140 mM HCO3-. High levels of HCO3- maintain secreted proteins such as mucins (all tissues) and digestive enzymes (pancreas) in a soluble and/or inactive state. HCO3- secretion is impaired in CF in all CFTR-expressing, HCO3--secreting tissues examined. The mechanism responsible for this critical problem in CF is unknown. Since a major component of HCO3- secretion in CFTR-expressing cells is mediated by the action of a Cl-/HCO3- exchanger (AE), in the present work we examined the regulation of AE activity by CFTR. In NIH 3T3 cells stably transfected with wild type CFTR and in HEK 293 cells expressing WT and several mutant CFTR, activation of CFTR by cAMP stimulated AE activity. Pharmacological and mutagenesis studies indicated that expression of CFTR in the plasma membrane, but not the Cl- conductive function of CFTR was required for activation of AE. Furthermore, mutations in NBD2 altered regulation of AE activity by CFTR independent of their effect on Cl- channel activity. At very high expression levels CFTR modified the sensitivity of AE to 4,4'-diisothiocyanatostilbene-2, 2'-disulfonate. The novel finding of regulation of Cl-/HCO3- exchange by CFTR reported here may have important physiological implications and explain, at least in part, the impaired HCO3- secretion in CF.

3T3 Cells↗
PubMed1999

Nephroprotective effect of betamipron on a new carbapenem, DA-1131, in rabbits.

The nephroprotective effect of betamipron (200 mg/kg) was evaluated after intravenous administration of a high dose of DA-1131 (200 mg/kg) to rabbits. Extensive tubular necrosis was observed without betamipron but the necrosis was not observed with betamipron at 8 h after intravenous administration of DA-1131 based on kidney microscopy. By treatment with betamipron, the amounts and tissue to plasma (T/P) ratios of DA-1131 in renal cortex and whole kidney decreased significantly (65-91% decrease) at both 30 min and 2 h after intravenous administration of the drug to rabbits. This indicated that the accumulation of DA-1131 in rabbit renal cortex and whole kidney was inhibited by betamipron. This resulted in significantly greater percentages of intravenous DA-1131 excreted in urine as unchanged drug, 60.9 versus 40.1%, and significantly faster renal clearance (Cl(r)) of DA-1131 (6.10 versus 3.22 mL/min/kg) by treatment with betamipron. By treatment with betamipron, the amounts and T/P ratios of DA-1131 in renal cortex and whole kidney decreased significantly from 30 min and the renal function remained intact at 8 h after intravenous administration of DA-1131. The above data suggested that the nephroprotective effect of betamipron was fast and persisted for a long period of time in rabbits.

Alanine↗
PubMed1999

Gender differences in pharmacokinetics and pharmacodynamics of azosemide in rats.

Gender differences in pharmacokinetics and pharmacodynamics of azosemide were evaluated after intravenous, 10 mg kg(-1), and oral, 10 mg kg(-1), administration to male and female rats. After intravenous administration to male rats, the percentages of intravenous dose of azosemide recovered from entire gastrointestinal tract at 24 h (13.2 versus 3.93%) was significantly greater than those in female rats. In male rats, the nonrenal clearance of azosemide tended (p<0.066) to be faster and kidney weight tended (p<0.068) to be greater than those in female rats. After oral administration of azosemide to male rats, the 8-h urinary excretion of potassium (0.395 versus 0.766 mmol g(-1) kidney) and 8-h kaluretic efficiency (55.9 versus 284 mmol mg(-1)) decreased significantly compared with female rats.

Administration, Oral↗
PubMed1999

Stability, blood partition and plasma protein binding of ipriflavone, an isoflavone derivative.

It is generally recognized that the partition between plasma and blood cells, the immediate centrifugation of blood samples after collection for the measurement of 'true' in vivo concentrations and free drug concentrations in plasma are important determinants of the pharmacokinetics and/or pharmacodynamics of drugs. Therefore, the stability, blood partition between plasma and blood cells, and factors influencing the binding of ipriflavone to 4% human serum albumin (HSA) using an equilibrium dialysis technique were evaluated. Ipriflavone was unstable in rat liver homogenate and various pH solutions ranging from 1 to 13, except pH 8, rat blood and plasma and human plasma when incubated in a water-bath shaker for 24 h kept at 37 degrees C and at a rate of 50 oscillations/min. The recoveries of spiked amounts of ipriflavone at 24 h pH solutions ranging from 1 to 12 were 67.0, 78.1, 87.9, 89.6, 84.2, 87.4, 85.5, 99.3, 88.0, 76. 6, 79.4 and 81.5%, respectively. Ipriflavone was very unstable in pH 13 solution; only 0.814% of ipriflavone was recovered after 30 min incubation. Ipriflavone was stable for up to 3 h incubation in human gastric juices. Ipriflavone reached equilibrium fast (within 30 s of being mixed manually) between plasma and blood cells and the equilibrium plasma/blood cells partition ratios were independent of the initial rabbit blood concentrations of ipriflavone: 0.2, 2, and 10 microg/mL; the values were in the range of 0.900-2.45. The binding of ipriflavone to 4% HSA was 96.6+/-0.407% at ipriflavone concentrations ranging from 2 to 100 microg/mL, but it was dependent on HSA concentrations (0.5-6%), incubation temperature (4, 22 and 37 degrees C), 'the buffer' pHs (5.8, 6.4, 7.0, 7.4 and 8.0), and addition of salicylic acid (150-300 microg/mL) and sulphisoxazole (100-300 microg/mL). However, the binding was independent of buffers containing various concentrations of chloride ion (0-0.546%), glucose (0 and 5%), alpha-1-acid glycoprotein (0-0.32%) and heparin (0-40 U/mL), and addition of its metabolites (M1 and M5, 5 microg/mL).

Animals↗
PubMed1999

Pyogenic hepatic abscesses: MRI findings on T1- and T2-weighted and serial gadolinium-enhanced gradient-echo images.

The purpose of this study was to determine the magnetic resonance imaging (MRI) features of pyogenic hepatic abscesses on T1-weighted, T2-weighted, and serial gadolinium (Gd)-enhanced T1-weighted spoiled gradient-echo (SGE) images including images acquired in the immediate, intermediate, and late phases of enhancement. The MRI studies of 20 patients with pyogenic liver abscesses were retrospectively reviewed. All patients were examined on 1.5 (n = 19) and 1.0 (n = 1) T MR scanners. MR studies included T1-weighted, T2-weighted, and serial Gd-enhanced SGE images. The following determinations were made: signal intensity of the abscess cavity and perilesional liver tissue, and the presence of internal septations, layering material, or air in the abscess cavity. The pattern of enhancement of the abscess wall, internal septae and peri-abscess liver were evaluated on serial Gd-enhanced SGE images. A total of 53 abscesses were observed in the 20 patients. Fortyeight abscesses were hypointense on T1-weighted and hyperintense on T2-weighted images. Internal septations were present in four abscesses. Lower signal intensity material was observed in a dependent location on T2-weighted images in one abscess. Signal void foci of air located on the nondependent surface was observed in two abscesses. Two other abscesses contained signal void air that occupied the entire abscess cavity, observed on all imaging sequences. On serial gadolinium-enhanced images, all abscesses revealed early enhancement of the wall, which persisted with negligible change in degree of enhancement or thickness on delayed images. Abscess walls ranged in thickness from 2 to 5 mm. Internal septations ranged in thickness from 2 to 3 mm. Abscess walls and septations were relatively uniform in thickness with no evidence of focal nodularity. Periabscess liver tissue was mildly hypointense on T1-weighted and mildly hyperintense on T2-weighted images in 20 lesions, which were either circumferential (n = 12) or wedge-shaped (n = 8). All these regions enhanced more than the remainder of the liver on immediate post-gadolinium images and remained relatively hyperintense on late phase images. Periabscess liver parenchyma was isointense on both T1- and T2-weighted images in 18 lesions, and in these lesions wedge-shaped subsegmental (n = 6) or segmental (n = 12) enhancement was observed on immediate gadolinium-enhanced images, which faded to isointensity on intermediate phase images. No perilesional signal changes and enhancement difference was observed in 15 lesions. Characteristic features of abscesses include: intense mural enhancement on early gadolinium-enhanced images, which persists with negligible change in thickness and intensity on later post-gadolinium images, and the presence of periabscess increased enhancement on immediate post-gadolinium images. These MRI features may help to distinguish abscesses from other focal liver lesions during differential diagnosis.

Contrast Media↗
PubMed1999

A survey of Blastocystis in domestic chickens.

The prevalence and morphology of Blastocystis in fresh faecal material from 227 domestic chickens was investigated. A very high prevalence of infection (approximately 95%) was found in chickens from four of the five commercial farms studied. Extremely high numbers of Blastocystis were found in the majority of samples. Blastocystis cells showed considerable variation in size, ranging from approx. 3 microm to approx. 120 microm in diameter. This size range is more extreme than those previously recognised for the organism from chickens. All chickens from one farm appeared free of Blastocystis infection. Most Blastocystis cells appeared to be the vacuolar form, although the shape of the cells and the appearance of the central vacuole contents varied considerably within and among faecal samples. Nuclei showed "spots" of electron-opaque material, generally arranged as a band within the nuclei. Multiple individual cysts within a single outer fibrillar layer were found in addition to single cysts without an encompassing fibrillar layer.

Animals↗
PubMed1999

Seroepidemiology of Helicobacter pylori infection in a Jamaican community.

We researched epidemiologic associations between environmental and demographic factors and prevalence of Helicobacter pylori infection in a suburban Jamaican community. Using a clustered sampling technique, 22 domestic yards enclosing 60 separate households were randomly selected from a local community. All household members (n = 346) were invited to participate following informed consent; the overall compliance rate was 58.9%. A commercial enzyme immunoassay (HMáCAP) was used to detect IgG antibodies raised against H. pylori. Environmental and demographic information was obtained by questionnaire. The seroprevalence of H. pylori was 69.9% (n = 202). Analysis of the independent variables revealed three major components: Component 1 described, collectively, good personal hygiene and sanitation, indoor water supply and absence of straying animals in the peridomestic area; Component 2 included older age, good personal hygiene and large yard size; Component 3 the presence of domestic animals (cats and dogs) and, again, large yard size. These three complexes explained 42.2% of the variability in the data set. Logistic regression showed that Components 2 and 3 were independently associated with H. pylori seropositivity, indicating that a combination of demographic, environmental and zoonotic factors is involved in the spread of H. pylori infections at the tropical community level.

Adolescent↗
PubMed1999

Sinus histiocytosis (Rosai-Dorfman disease) clinically limited to the skin.

Sinus histiocytosis with massive lymphadenopathy (SHML) is an idiopathic proliferation of unique histiocytes that have vesicular nuclei and voluminous pale cytoplasm, often with emperipolesis. Pure cutaneous involvement is very rare. We describe a patient with SHML limited to the skin whose lesion has spontaneously regressed. A 35-year-old Korean male visited the Department of Dermatology due to facial rash for 2 months. A 3 x 3.5 cm-sized well-demarcated dark erythematous nontender plaque was noted on the right cheek. Skin biopsy showed dense, nodular infiltrates of histiocytes with abundant cytoplasm and vesicular nuclei rimmed by lymphoplasma cell aggregates throughout the upper and mid-dermis. The histiocytes were immunohistochemically positive for S-100 protein and CD68, but negative for CD1a. Laboratory tests and a thorough physical examination revealed no abnormalities. These findings suggested that this was a case of SHML clinically limited to the skin. The skin lesion was initially resistant to steroid therapy, but began to regress 10 months after the onset without further treatment.

Adult↗
PubMed1999

Distally based free flap using retrograde arterial flow.

In lower extremity injuries, the major arteries are frequently damaged and the remaining artery may be located in the traumatized zone. Recently, vascular anastomoses to recipient vessels distal to the zone of injuries have been advocated in the lower limb reconstruction and flaps by using retrograde arterial flow are well established in island flaps. Twelve patients with soft-tissue defects below the knee underwent lower extremity reconstruction with distally based free flaps by using retrograde arterial flow. For assessment of retrograde flow, the intraoperative retrograde arterial pressure was quantitated and compared with the systolic blood pressure taken at the same time (n = 6). In this study, suitable candidates had been chosen with pulsatile retrograde flow from the distal cut end of the vessel because patients without pulsatile flow had recorded diastolic and systolic retrograde arterial pressures that were very low. Four different free flaps were used. All flaps were successful. Retrograde flow anastomosis could not interrupt the major blood vessels, which were essential for survival of the distal limb, the compromise of fracture or wound healing might be prevented. In cases in which arteriography demonstrates significant vascular flow interruption within the zone of injury, retrograde arterial anastomosis may also be a candidate.

Adolescent↗