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Biomedical subjects

S L Yang

Publications and source records attributed to S L Yang.

At least 109 records · Page 6Linked to original sources

Successful donation and transplantation of multiple organs from a victim of cyanide poisoning.

Demand for viable human organs for transplantation continues to exceed the supply. To expand supply, the criteria for identification and management of suitable donors must continue to evolve. Poisoned patients are often excluded as potential organ donors due to perceived risks of transmittable agents and/or physiologically compromised organs. In this report, a patient succumbed after an intentional ingestion of cyanide and multiple pharmaceuticals. Donor organ viability was determined by lack of significant injury beyond the central nervous system. Following standard procurement procedures, the heart, liver, corneas, 16 skin grafts and 16 bone grafts were deemed suitable and successfully transplanted. All organ recipients were doing well eight months post transplantation. The focus of procurement personnel should be on tissue injury and not on the mere presence of clinical effect of a toxic agent. With the paucity of organs available, poison centers need to be cognizant of this dilemma when faced with a toxicologically compromised potential organ donor.

Adult↗

Electrophysiologic properties of isolated adult cardiomyocytes from septic rats.

In this study, we examined the effect of sepsis on the electrical properties of isolated ventricular myocytes. Sepsis was induced by cecal ligation and puncture (CLP). The control rats were sham-operated. Membrane potentials and ionic currents in isolated cardiac myocytes were measured by the tight-seal, whole-cell patch-clamp technique. The results show that the resting membrane potentials of heart cells were significantly lower in the septic group (18 hr post-CLP) than those in the control group. However, there was no significant difference in action potential duration of 50% and 90% repolarization between the two groups of cells. In voltage-clamp experiments, isoproterenol (10 nM), a beta-adrenergic agonist, caused an increase in L-type calcium current (ICa,L) in a similar magnitude in myocytes isolated from the control and septic rats. Furthermore, isoproterenol failed to modify the time constants for ICa,L inactivation and the overall shape of current-voltage relationship for both groups of cells. These results indicate that formation of a G omega seal and subsequent tight-seal whole-cell recording with patch-clamp technique can be performed in heart cells derived from CLP-induced septic rats, and that septic rat heart is capable of responding effectively to beta-adrenergic stimulation.

Animals↗

[Lysosomal enzyme activity of cultured fetal cells in Chinese and its clinical application].

Lysosomal storage diseases (LSD) are caused by deficient activity of specific lysosomal enzymes. Early diagnosis and selective termination is still the trend of therapy. The purpose of this study was to establish an assay system and investigate the reference range of lysosomal enzyme activity of cultured fetal cells in the Chinese population. Seventy amniotic fluid and 9 chorionic villi samples were collected and cultured in this study. Enzyme activity assay was done by synthesized 4-Mu-binded substrates. The activity was expressed as nmol/mg protein/hour. In cultured amniotic cells, the results showed 14-138 of alpha-glucosidase, 8-133 of alpha-galactosidase, 32-470 of alpha-mannosidase, 101-1121 of alpha-fucosidase, 106-1321 of beta-galactosidase, 15-268 of beta-glucosidase, 11-279 of beta-glucuronidase, 101-1193 of Hexosaminidase A, and 886-6204 of N-acetyl-alpha-glucosaminidase. In cultured chorionic villi samples, it showed 22-335 of alpha-glucosidase, 31-230 of alpha-galactosidase, 47-250 of alpha-mannosidase, 35-218 of alpha-fucosidase, 49-934 of beta-galactosidase, 34-329, of beta-glucosidase, 57-379 of beta-glucuronidase, and 328-3412 of Hexosaminidase A. The enzyme activity was not correlated with the gestation age when sample was obtained. Furthermore, there was no statistical significance among the range of amniotic cells, chorionic villi samples, skin fibroblasts and peripheral leukocytes for each enzyme studied. It is suggested that the synthesis of lysosomal enzymes has been mature since the early fetal state, and the samples obtained as early as 8 weeks of gestation age can be used for early diagnosis of lysosomal storage diseases.

Cells, Cultured↗

Combined team effort in liver procurement and transplantation: 5-year experience of one organ procurement organization.

Combined team effort in liver procurement and transplantation has proven to be very advantageous as compared to same team effort in our experience. The incidence of primary nonfunction was almost identical between the two groups. We recommend that a combined team effort be used in liver transplantation to conserve the resources and ensure the safety of the transplant team. The practice of only a single local recovery team for all abdominal organ procurement should be encouraged.

Delaware↗

Organ donation and referrals among African-Americans.

Solid organ referrals for AA have been growing since 1989. One third of AA referrals are in the 16 to 30 year age group with the majority of these males suffering traumatic injury. AA donors have increased from 11 in 1989 to 32 in 1992. This is in spite of AA and minority consent rates that are 20% to 30% below that of the CAUC group. This gap represents a vital opportunity for improvement. Increased education to minority groups with a message that considers the ethnic and cultural diversity of the minority community coupled with an increase in the number of minority transplant coordinators should help to increase the number of AA referrals and donors.

Adolescent↗

[Kinetic studies of protein kinase A in rat liver during early sepsis].

Sepsis-induced glucose dyshomeostasis has been characterized by an initial hyperglycemia followed by a progressive hypoglycemia. It is well known that the liver plays a predominant role on regulating the homeostatic level of blood glucose. Furthermore, recent studies indicate that protein kinase A, activated by c-AMP, contributes to the role of glycagon in glucogenolysis and glyconeogenesis. Kinetic studies of protein kinase were completed. During late sepsis, in order to further understand the pathophysiology of hepatic glucose disturbances during sepsis. This study investigates the role of protein kinase A in the liver regulating carbohydrate metabolism during early sepsis. The work was performed by using an animal septic model, induced by cecal ligation and puncture (CLP) operation. Through the measurement of blood sugar, a two phase change in sugar level was found. That is, blood sugar significantly increased at 4.5 hrs after CLP operation (p < 0.05) and then significantly decreased at 18 hrs (p < 0.01). In the kinetic studies of protein kinase A, the results showed that, during early sepsis, the activities of both type I (eluted at low ionic strength) and type II (eluted at high ionic strength) protein kinase A were unchanged. Moreover, the kinetic parameters, Vmax and S0.5, of protein kinase A showed no significant difference between two groups. As such, it is suggested that hyperglycemia during early sepsis is not connected to the regulation of protein kinase A.

Adenosine Triphosphate↗

Regional mapping of prion proteins in brain.

Scrapie is characterized by the accumulation of a protease-resistant isoform of the prion protein PrPSc. Limited proteolysis and chaotropes were used to map the distribution of PrPSc in cryostat sections of scrapie-infected brain blotted onto nitrocellulose membranes, designated histoblots. Proteolysis was omitted in order to map the cellular isoform of the prion protein (PrPC) in uninfected brains. Compared with immunohistochemistry, histoblots increased the sensitivity for PrPSc detection and showed different patterns of PrPSc accumulation. In Syrian hamsters with Sc237 scrapie, the most intense PrPSc signals occurred in sites with relatively little PrPC, suggesting that aberrant localization of prion protein may be an important feature in the pathogenesis of prion diseases. Immunostaining of PrPSc in white-matter tracts suggested that prions spread along neuroanatomical pathways. PrPSc immunostaining in histoblots was quantitated by densitometry, permitting assessment of the extent of PrPSc accumulation within specific structures. Histoblots were also useful in localizing PrPCJD and beta/A4-amyloid peptide in the brains of patients with Creutzfeldt-Jakob disease and Alzheimer disease, respectively.

Animals↗

Replication of distinct scrapie prion isolates is region specific in brains of transgenic mice and hamsters.

Scrapie prions are composed largely, if not entirely, of PrPSc molecules. The prion isolates Sc237 and 139H exhibit markedly different incubation times in Syrian, Armenian, and Chinese hamsters, as well as in transgenic (Tg) 81 mice expressing Syrian hamster PrP (SHaPrP). Repassage of prions from transgenic mice or Chinese hamsters into Syrian hamsters revealed that the original properties of the prion isolates are retained. When Syrian hamsters were first inoculated with 139H prions and subsequently challenged with Sc237 prions, the incubation period was determined by the faster Sc237 isolate. Regional mapping studies demonstrated different kinetics and patterns of PrPSc accumulation for Sc237 and 139H prions in the brains of Syrian hamsters as well as Tg(SHaPrP)7 mice. That distinct prion isolates induce different region-specific accumulations of PrPSc in brain suggests a novel mechanism for propagation of isolates whereby they replicate in particular sets of neurons. The prion isolates could be targeted to specific CNS cells by differing conformations of PrPSc, post-translational modifications of PrPSc such as Asn-linked glycosylation, or an as yet undetected macromolecule complexed with PrPSc in the prion.

Animals↗

15NH4+ excretion test: a new method for detection of Helicobacter pylori infection.

A noninvasive test for the detection of Helicobacter pylori infection that uses [15N]urea as a tracer has been established. The principle the test is based on is the strong urease activity of H. pylori. After oral ingestion, [15N]urea is broken down into ammonia and carbon dioxide by H. pylori urease in the stomach. The ammonia is absorbed into the blood and excreted in the urine. The amount of [15N]urea, reflecting the magnitude of H. pylori infection, is evaluated by measuring the abundance and excretion rate of 15N in ammonia in the urine. Thirty-six patients were examined in our study. The 15N excretion rates in urine ammonia of patients who were H. pylori positive were significantly higher than those of H. pylori-negative patients (P less than 0.05). Twenty-three patients were H. pylori positive by Gram stain and culture. The sensitivity of the 15NH4 excretion test compared with these techniques was 96%, and no false positives were obtained. The 15NH4+ excretion rates of 13 H. pylori-negative subjects were all in the normal range (less than 0.3%). This method is a simple, precise, highly sensitive, noninvasive, nonradioactive test. It could be used for diagnosis as well as for the followup of patients receiving H. pylori eradication therapy, especially children and pregnant women. It could also be used in epidemiological investigation of H. pylori infection in a general population.

Adult↗

[Kinetic studies of protein kinase A in rat liver during late sepsis].

The covalent modification of receptor proteins via phosphorylation and dephosphorylation is one of the principal mechanisms controlling carbohydrate metabolism and is known to be regulated by various protein kinases. Recent studies indicated that many hormones may exert their effects on cellular metabolism by regulating intracellular c-AMP levels and by activating a c-AMP dependent protein kinase, i.e., protein kinase A. The metabolic disturbances during sepsis are characterized by an initial hyperglycemia followed by a progressive hypoglycemia and a depletion of hepatic glycogen content. The latter is coupled with a slowdown in glycogenesis, an accelerated glycogenolysis, and a depression in gluconeogenesis in the liver. Since the liver is the major organ that regulates the homeostatic level of blood glucose, it is conceivable that the sepsis-induced glucose dyshomeostasis might be mediated by changes in protein kinase activity and the kinetic characteristics of enzymes. The present experiment was designed to study the correlation between protein kinase A and the pathophysiology of hepatic glucose dyshomeostasis during sepsis. Sepsis was induced in rats by cecal ligation and puncture (CLP). Late sepsis occurred 18 hours after CLP. Protein kinase A was extracted from the rat livers by acid precipitation and ammonium sulfate fractionation, and then partially purified by DEAE-cellulose. The results show that in the late sepsis, type-I protein kinase A (eluted at low ionic strength) activity was significantly decreased by 34-52% (P < 0.01). The kinetic parameters such as Vmax's for ATP, histone, and c-AMP were also significantly decreased from the control values of 6.1 +/- 0.9, 5.4 +/- 0.8, and 5.1 +/- 1.9 nmoles/mg.min. to 3.6 +/- 0.5, 2.8 +/- 0.3, and 2.5 +/- 0.5 nmoles/mg.min., respectively. Analysis using Hill's equation indicates that the S0.5 and n (Hill coefficient) values of the various substrates and activators for type-I protein kinase A remained unchanged. In the case of type-II protein kinase A (eluted at high ionic strength), the Vmax, S0.5, and n values for ATP, histone, and c-AMP were unchanged during late sepsis. The results of the present study indicate that the activities and kinetic characteristics of type I protein kinase A in rat liver are modified during late sepsis. Since protein kinase A is known to regulate glucose metabolism through adrenergic receptor mediation, these findings may have a pathophysiological significance in the understanding of hepatic glucose dyshomeostasis during sepsis.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

A possible role of heat shock proteins in human sperm motility.

The testicular spermatogenic stem and seminiferous tubular cells selectively synthesize heat shock proteins (Hsps) during heat stress. Hsps, synthesized from testicular cells and leukocytes, are identical in molecular masses as well as chemical properties. In this study, we induced the Hsps from leukocytes and investigated their in vitro effects on human sperm motility. Semen samples were divided into two parts, washed and unwashed. The whole blood was heated in 43 degrees C for 15 minutes for induction of Hsps. A trans-membrane migration method was used to examine the effect of heated blood plasma on human sperm motility. The main heat-induced proteins of leukocytes were detected by 2-D electrophoresis and Coomassie blue stain. Leukocytes treated by heat produced a large amount of Hsp72 and Hsp80, while only a small amount was observed in that of non-heated leukocytes. The heated blood plasma inhibited motility of washed sperm in a manner that was dose-dependent. In the presence of seminal plasma fluid, however, the inhibitory effects of heated plasma on human sperm motility could not be observed. It was concluded that the heat-induced substance(s) from leukocytes, which being highly possible the Hsps, interfered the mobility of wash human sperm and the inhibition might be antagonized by seminal plasma.

Heat-Shock Proteins↗

[The effect of monosodium L-glutamate (MSG) treatment during neonatal period on the pineal function in young female rats].

The purpose of this study is to examine whether monosodium L-glutamate (MSG) influences the function of pinealocytes in female rats. MSG was administered at a does of 4 mg/g body weight to rat pups on days 1 and 3 postnatally. As the rats matured, sexual receptivity as well as proceptivity were observed by testing the lordosis quotient (LQ), rejection quotient (RQ) and solicitation. Pineal function was estimated by measuring the serum level of melatonin and the activity of protein kinase A (PKA) in the pineal gland. Blood samples were withdrawn at light and dark phases for four consecutive days, respectively. The serum concentration of melatonin was determined by radioimmunoassay and the data of each phase were pooled together. The results showed that both the receptivity and proceptivity of MSG-treated female rats were significantly lower than that of the control ones. Melatonin levels during the dark phase were significantly higher than those during the light phase in both control and MSG-treated groups, but whenever in the light or dark phase, the levels of melatonin were the same in both groups. There was no significant difference in PKA activity in the pineal gland between the control and MSG-treated group. These results indicate that MSG used as a neurotoxin to induce hypogonadal status affected neither the PKA activity of pineal gland nor the serum level of melatonin in young female rats. Thus, the decrease in receptivity and proceptivity of MSG-treated female rats was not caused by the alteration of pineal function.

Animals↗

Paradoxical shortening of scrapie incubation times by expression of prion protein transgenes derived from long incubation period mice.

Prolonged incubation times for experimental scrapie in I/LnJ mice are dictated by a dominant gene linked to the prion protein gene (Prn-p). Transgenic mice were analyzed to discriminate between an effect of the I/LnJ Prn-pb allele and a distinct incubation time locus designated Prn-i. Paradoxically, 4 independent Prn-pb transgenic mouse lines had scrapie incubation times shorter than nontransgenic controls, instead of the anticipated prolonged incubation periods. Aberrant or overexpression of the Prn-pb transgenes may dictate abbreviated incubation times, masking genuine Prn-p/Prn-i congruence; alternatively, a discrete Prn-i gene lies adjacent to Prn-p.

Animals↗

The role of the posterior cricoarytenoid muscle in phonation: an electromyographic investigation in dogs.

The activity of the posterior cricoarytenoid muscle during respiration has been well investigated electromyographically. Its activity during phonation, however, has not been studied systematically. We, therefore, focused our attention on the phonatory activity of the posterior cricoarytenoid muscle to confirm whether it also contracts during phonation, as has been reported by some researchers. In our series of 12 adult dogs, the posterior cricoarytenoid was active in 11 dogs and inactive in one dog during phonation. Our present study also showed that the posterior cricoarytenoid activity was stronger for phonation than for inspiration in 6, stronger for inspiration than for phonation in 3, and the same for phonation and inspiration in 2 dogs. The results obtained from the present electromyographic evaluation demonstrated that the posterior cricoarytenoid is activated during phonation. The authors believe that the posterior cricoarytenoid muscle has phonatory function and that the phonatory effect of this muscle on the vocal cord may play an important role in precise glottis control.

Animals↗

Current status of biotechnology in China.

There has been increasing interest over past decade in exploring the possibility of using new biotechnology to produce new products and to improve the old productive process. The researches and applications of genetic engineering, cell fusion, mutagenesis, cell and enzyme immobilization in enzyme, antibiotic, vitamins, steroid, amino acid, organic acid, solvent, food and brewage industries is reviewed.

Anti-Bacterial Agents↗

An efficient site-directed mutagenesis using polymerase chain reaction.

We report here a simple and efficient method for site-directed mutagenesis using polymerase chain reaction (PCR). In constructing a new expression plasmid for the EcoRI restriction gene, we made two point mutations. While one created a new SalI site prior to the SD sequence, the other replaced Glu144 with Lys. A 1.5 kb SalI-PstI fragment isolated from pER101 was used as the template. Two 25 mer oligonucleotide primers containing the desired mutations were synthesized and used to direct PCR amplification with Taq DNA polymerase. About 0.5 microgram of the 0.49 kb fragment was obtained from 0.05 microgram of the 1.5 kb fragment by carrying out polymerase chain reaction for 30 cycles. As calculated theoretically, 99% of the product contained the desired mutations. The product was cloned into pUC19 using SalI and PstI, two of the transformed colonies were randomly chosen for sequence analysis, and both of them were shown to contain the desired mutations. Finally, the amplified fragment was cloned into pER304 to place the EcoRI (Lys144) gene directly under the control of the lambda PL promoter.

Base Sequence↗

Transgenetic studies implicate interactions between homologous PrP isoforms in scrapie prion replication.

Transgenic (Tg) mice expressing both Syrian hamster (Ha) and mouse (Mo) prion protein (PrP) genes were used to probe the mechanism of scrapie prion replication. Four Tg lines expressing HaPrP exhibited distinct incubation times ranging from 48 to 277 days, which correlated inversely with HaPrP mRNA and HaPrPC. Bioassays of Tg brain extracts showed that the prion inoculum dictates which prions are synthesized de novo. Tg mice inoculated with Ha prions had approximately 10(9) ID50 units of Ha prions per gram of brain and less than 10 units of Mo prions. Conversely, Tg mice inoculated with Mo prions synthesized Mo prions but not Ha prions. Similarly, Tg mice inoculated with Ha prions exhibited neuropathologic changes characteristic of hamsters with scrapie, while Mo prions produced changes similar to those in non-Tg mice. Our results argue that species specificity of scrapie prions resides in the PrP sequence and prion synthesis is initiated by a species-specific interaction between PrPSc in the inoculum and homologous PrPC.

Animals↗