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Biomedical subjects

S L Lin

Publications and source records attributed to S L Lin.

At least 19 recordsLinked to original sources

A Novel mRNA-cDNA interference phenomenon for silencing bcl-2 expression in human LNCaP cells.

The templates required for inducing posttranscriptional gene silencing (PTGS) effects have been investigated in human prostate cancer LNCaP cells. Transfection of a mRNA-cDNA hybrid construct was found to result in a relatively long-term interference of specific gene expression. Androgen-stimulated expression of bcl-2 has been reported to increase the tumorigenic and metastatic potentials of human prostate cancer LNCaP cells, as well as their resistance to many apoptotic stimuli. The addition of bcl-2 antisense oligonucleotides, however, restored apoptosis. Our studies demonstrate gene silencing effects of the mRNA-cDNA transfection that is similar to those of PTGS/RNAi in this in vitro prostate cancer cell model. A potential RNA-directed RNA polymerase activity was also detected which is alpha-amanitin-sensitive. These findings indicate that a novel gene silencing system may exist in mammalian cells.

Base Sequence↗

A unique sialidase that cleaves the Neu5Gcalpha2-->5-O(glycolyl)Neu5Gc linkage: comparison of its specificity with that of three microbial sialidases toward four sialic acid dimers.

We found that the hepatopancreas of oyster, Crassostrea virginica, contained a sialidase capable of releasing Neu5Gc from the novel polysialic acid chain (-->5-O(glycolyl)Neu5Gcalpha2-->)n more efficiently than from the conventional type of polysialic acid chains, (-->8Neu5Acalpha2-->)n, or (-->8Neu5Gcalpha2-->)n. We have partially purified this novel sialidase and compared its reactivity with that of microbial sialidases using four different sialic acid dimers, Neu5Gcalpha2-->5-O(glycolyl)Neu5Gc (Gg2), Neu5Acalpha2-->8Neu5Ac (A2), Neu5Gcalpha2-->8Neu5Gc (G2), and KDNalpha2-->8KDN (K2) as substrates. Hydrolysis was monitored by high performance anion-exchange chromatography with a CarboPac PA-100 column and pulsed amperometric detection, the method by which we can accurately quantitate both the substrate (sialiac acid dimers) and the product (sialic acid monomers). The oyster sialidase effectively hydrolyzed Gg2 and K2, whereas A2 and G2 were poor substrates. Neu5Ac2en but not KDN2en effectively inhibited the hydrolysis of Gg2 by the oyster sialidase. Likewise, the hydrolysis of K2 by the oyster sialidase was inhibited by a cognate inhibitor, KDN2en, but not by Neu5Ac2en. Using the new analytical method we found that Gg2 was hydrolyzed less efficiently than A2 but much more readily than G2 by Arthrobacter ureafaciens sialidase. This result was at variance with the previous report using the thiobarbituric acid method to detect the released free sialic acid [Kitazume, S., et al. (1994) Biochem. Biophys. Res. Commun. 205, 893-898]. In agreement with previous results, Gg2 was a poor substrate for Clostridium perfringens sialidase, while K2 was refractory to all microbial sialidases tested. Thus, the oyster sialidase is novel and distinct from microbial sialidases with regards to glycon- and linkage-specificity. This finding adds an example of the presence of diverse sialidases, in line with the diverse sialic acids and sialic acid linkages that exist in nature. The new sialidase should become useful for both structural and functional studies of sialoglycoconjugates.

Animals↗

Laparoscopic closure of patent canal of Nuck for female indirect inguinal hernia.

STUDY OBJECTIVE: To determine the efficacy of concomitant laparoscopic closure of the patent canal of Nuck in treatment of indirect inguinal hernias in women. DESIGN: Prospective clinical trial (Canadian Task Force classification II-2). SETTING: University-associated tertiary care referral center. PATIENTS: Six women scheduled for laparoscopy for gynecologic indications. Intervention. Concomitant simple closure of the patent canal of Nuck from the peritoneal cavity during laparoscopic surgery. MEASUREMENTS AND MAIN RESULTS: Inguinal hernias were repaired in all six patients, with no recurrence. CONCLUSION: l Laparoscopic closure of the patent canal of Nuck is an easy, safe, and effective treatment of indirect inguinal hernias in women. (J Am Assoc Gynecol Laparosc 8(1):143-146, 2001)

Adult↗

External biliary drainage plus bile acid feeding is not equal to internal drainage in preserving the cellular immunity following prolonged obstructive jaundice.

This study investigates the importance of intestinal bile flow in cellular immunity. Sprague-Dawley rats undergoing bile duct ligation (BDL) and sham ceiliotomy (Sham) for 14 and 21 days were investigated. Experimental animals following BDL were further divided into an external drainage (ED) group, an ED group with rat chow mixed with 2:2:1 cholic acid, chenodeoxycholic acid, and deoxycholic acid (ED + BF), and an internal drainage (ID) group. Fourteen days later, they were killed and analyzed for spleen lymphocytic [3H] thymidine uptake (LHU) under mitogen stimulation with phytohemagglutinin, blood biochemistry, hemogram, and liver pathology. In the 14-day BDL experiment, LHU and serum albumin level were decreased in the BDL group (P < 0.05). After drainage, they were not significantly different among sham, ED, ED + BF, and ID groups. In the 21-day BDL experiment, the red cell volume was decreased (P < 0.05). After drainage, the ED, ED + BF, and ID groups still had a significantly lower LHU than the sham group (P < 0.05). However, the ID group had higher LHU than the ED and ED + BF groups (P < 0.05). The ED + BF group had a slightly higher LHU than the ED group but not statistically significant. Liver pathology returned to normal after drainage in the 14-day BDL model. In contrast, the 21-day BDL group had prominent periportal necrosis and developed periportal fibrosis after drainage. The present study reveals the duration of BDL determines the severity of hepatic damage. In the 14-day BDL groups, all kinds of drainage completely reverse the impaired liver function and cellular immunity. In the 21-day BDL group, 14-day drainage is inadequate for recovery because irreversible pathological changes are found. The reversal of cellular immunity in ID is better and faster, because it provides a better hepatic functional, nutritional, and hematological recovery besides the presence of primarily secreted bile acids.

Animals↗

Influence of erythropoietin treatment on gonadotropic hormone levels and sexual function in male uremic patients.

OBJECTIVE: The purpose of this study was to evaluate whether levels of sex hormones and sexual function differ in renal failure patients with and without uremia and the effect of treatment with recombinant human erythropoietin (rhuEPO). MATERIAL AND METHODS: Fifteen males with chronic renal failure who were not receiving hemodialysis and 25 male renal failure patients with uremia who were undergoing hemodialysis were enrolled before and after rhuEPO therapy. Fifteen male volunteers matched for age and weight were also studied. Levels of various blood biochemicals were measured in all patients before and 1 week after rhuEPO treatment. Sexual function was also studied in all patients before and 6 months after rhuEPO treatment. RESULTS: The control group had significantly higher levels of testosterone (6.21 +/- 1.21 ng/ml) and hematocrit (Hct) (43.2 +/- 2.1%) and significantly lower levels of prolactin (5.27 +/- 1.21 ng/ml), follicular-stimulating hormone (FSH) (7.51 +/- 2.36 mIU/ml) and leutinizing hormone (LH) (4.23 +/- 2.10 mIU/ml) than the two patient groups (p < 0.05 for all comparisons). Patients with renal failure only had significantly lower levels of testosterone and Hct (2.54 +/- 0.53 ng/ml and 21.4 +/- 1.4%, respectively) than those with uremia (3.65 +/- 0.52 ng/ml and 24.3 +/- 2.5%, respectively; p < 0.001 for both comparisons). After rhuEPO therapy, the testosterone and Hct levels of the two patient groups did not reach the level of the control subjects (p < 0.05 for both comparisons). Similarly, the levels of prolactin, FSH and LH were significantly higher in both patient groups than those of control subjects after rhuEPO therapy (p < 0.001 for both comparisons). However, after rhuEPO therapy, significant increases in testosterone and Hct levels were found in both patient groups (p < 0.001 for both comparisons). Sexual function was also markedly improved in the hemodialysis patient group. While 20/25 (80%) male hemodialysis patients reported improved sexual function after rhuEPO treatment, only 3/15 (20%) chronic renal failure patients reported improvement. CONCLUSIONS: In patients with advanced uremia, rhuEPO therapy may result in improved gonadotropic hormone levels and sexual function. Good dialysis quality may contribute to the increase in the incidence of patients with better sexual function.

Erythropoietin↗

An ultrasensitive chemical method for polysialic acid analysis.

An ultrasensitive method for analysis of polysialic acid (polySia) chains, using fluorescence-assisted high-performance liquid chromatography was developed. The new method is a substantial improvement of our earlier method in which the reducing terminal Sia residues of a homologous series of oligo/polySia hydrolytically released during derivatization reaction were simultaneously labeled with a fluorogenic reagent, 1,2-diamino-4,5-methylenedioxybenzene (DMB) in situ. We first studied extensively the stability of oligo/polySia in the acid (0.02 M trifluoracetic acid) used for 1,2-diamino-4,5-methylenedioxybenzene derivatization under various conditions of reaction time and temperature, analyzing the hydrolytic products by high-performance anion exchange chromatography with pulsed electrochemical detection (HPAEC-PED). Then we optimized the reaction conditions to minimize degradation of the parent polySia while maintaining high derivatization rate. Using a DNAPac PA-100 column rather than a MonoQ column, baseline resolution of polySia peaks up to DP 90 with a detection threshold of 1.4 femtomol per resolved peak was achieved. The new method was used to analyze the degree of polymerization of a polySia-containing glycopeptide fraction derived from embryonic chicken brain, and the results were compared with those obtained by HPAEC-PED.

Animals↗

Extremely low sodium hypotonic rehydration solution for young children with acute gastroenteritis.

BACKGROUND: The clinical efficacy and safety of a low-sodium hypotonic oral rehydration solution (LSORS) was compared in a pilot study with that of a standard World Health Organization oral rehydration solution (STORS) in young children with acute diarrhea. METHODS: One hundred and seventeen boys aged 3 to 18 months with acute diarrhea were randomly assigned to groups and received low-sodium (sodium 28 mmol/L) hypotonic oral rehydration solution and standard oral rehydration solution (sodium 90 mmol/L). Outcomes of therapy such as stool volume, duration of diarrhea and fluid intake were recorded at 24 hours, 48 hours, and the discontinuation of disease. Serum electrolytes levels were calculated before and after therapy. RESULTS: The stool output (gm/kg) in the first 24 hours was 69.2 +/- 38.5 in the LSORS group versus 105 +/- 39.8 in the STORS group (p < 0.05), while the total stool output during the 48-hour period was 151.4 +/- 68.9 in the LSORS group versus 232 +/- 97.3 in the STORS group (p < 0.05). No significant (p > 0.05) reduction in duration of diarrhea was noted (72.3 +/- 25.5 hours versus 69.8 +/- 29.4 hours). Four patients in the LSORS group and 7 patients in the STORS were classified as treatment failure. Forty-four (80%) achieved successful treatment in the LSORS group and 34 (56.7%) in the STORS group (p = 0.07). CONCLUSION: LSORS is as safe as STORS in treating young children with acute gastroenteritis. No evidence of hyponatremia was found after patient received LSORS. LSORS afforded more efficacious therapy than STORS.

Acute Disease↗

Significance of cardiac troponin I and creatine kinase release after coronary intervention.

BACKGROUND: Cardiac troponin I is a highly sensitive and specific marker for early detection of myocardial injury. Whether it can be used to monitor myocardial injury after coronary intervention is uncertain. This study was designed to measure the cardiac troponin I and creatine kinase (CK) after coronary intervention and investigate their clinical significance. METHODS: We measured cardiac troponin I and CK levels before intervention and 4 hours, 8 hours, 12 hours and 24 hours after apparently successful coronary intervention in 106 eligible patients. Nine patients were excluded due to missing data. We also followed up the clinical outcome to record major cardiac events (MACE). RESULTS: The frequency of cardiac troponin I increase after coronary intervention was higher than that of CK increase (40.2% vs 8.2%). The frequency of cardiac troponin I increase in the stent group was significantly higher than that in the PTCA group (49.2% vs 21.9%, p < 0.001). The frequency of cardiac troponin I increase was also higher than that of CK increase in patients with in-hospital events (58.8% vs 14.7%). CONCLUSIONS: Cardiac troponin I is more sensitive than creatine kinase in detecting myocardial injury after coronary intervention. The incidence of cardiac troponin I increase is significantly higher in patients undergoing stenting than in patients treated with balloon angioplasty only. The cardiac troponin I increase is more highly correlated with in-hospital events than is creatine kinase.

Adult↗

Comparison of the APACHE II and APACHE III scoring systems in patients with respiratory failure in a medical intensive care unit.

BACKGROUND AND PURPOSE: This retrospective study compared the capability of the Acute Physiology and Chronic Health Evaluation (APACHE) II and APACHE III scoring systems to predict outcome and determined the independent predictors of survival in these scoring systems for patients with respiratory failure in a medical intensive care unit (ICU). MATERIALS AND METHODS: Seven hundred and eight patients with respiratory failure admitted to the medical ICU throughout a 9-year period were studied. Patients with an ICU stay of less than 24 hours, patients under 12 years of age, and burn and surgery patients were excluded. APACHE scores were calculated at 24 hours after admission. Student's t-test was used to compare the total APACHE scores of survivor and non-survivor groups. Multivariate logistic regression analysis was used to determine which variables were predictors of mortality. The discriminative power of APACHE scores to predict in-hospital mortality was studied by the area under the receiver operating characteristic curves of the APACHE II and APACHE III systems, respectively. RESULTS: Both systems showed a significant association between higher scores and higher mortality. The APACHE II system under-predicted the actual hospital mortality rate. The APACHE III systems had a higher discriminative power (area 0.7462) than the APACHE II systems (area 0.6856; p < 0.05). The independent predictors of survival as assessed by APACHE II and III systems were respiratory rate, arterial oxygen pressure, oxygen gradient between alveoli and artery, serum creatinine concentration, and the presence of neurologic abnormalities. CONCLUSIONS: The APACHE III systems has greater discriminative power than the APACHE II systems for predicting in-hospital mortality. The variables of oxygenation, mean artery pressure, respiratory rate, serum creatinine concentration, and Glasgow Coma Scale play important roles in predicting survival for patients with respiratory failure.

APACHE↗

High resistive index of the radial artery is related to early primary radiocephalic hemodialysis fistula failure.

Forty-nine patients who had received radiocephalic hemodialysis fistula construction were evaluated with duplex Doppler ultrasonography to characterize the Doppler indices of the feed radial arteries just proximal to the site of anastomosis. Forty-four patients had fistulas with good function, and 5 patients had fistulas with inadequate blood flow or thrombosis within 4 weeks after the operation. A preliminary study showed extensive variability in peak systolic velocity and end-diastolic velocity in the feed arteries. The resistive index dropped significantly 1 week after the operation and remained relatively constant over the following 5 weeks. In the success group, the mean resistive index measured 1 week after operation was 0.40+/-0.06. It was higher than that of the failure group (mean resistive index: 0.52+/-0.06). Among patients with well-functioning fistulas, diabetic patients had higher resistive indices than did non-diabetic patients (0.44+/-0.04 vs. 0.37+/-0.06). Our results suggest that a higher resistive index of the feed artery is closely related to early autogenous primary hemodialysis fistula failure.

Adult↗

Crystal structure of the oligomerization domain of NSP4 from rotavirus reveals a core metal-binding site.

During the maturation of rotaviral particles, non-structural protein 4 (NSP4) plays a critical role in the translocation of the immature capsid into the lumen of the endoplasmic reticulum. Full-length NSP4 and a 22 amino acid peptide (NSP4(114-135)) derived from this protein have been shown to induce diarrhea in young mice in an age-dependent manner, and may therefore be the agent responsible for rotavirally-induced symptoms. We have determined the crystal structure of the oligomerization domain of NSP4 which spans residues 95 to 137 (NSP4(95-137)). NSP4(95-137) self-associates into a parallel, tetrameric coiled-coil, with the hydrophobic core interrupted by three polar layers occupying a and d-heptad positions. Side-chains from two consecutive polar layers, consisting of four Gln123 and two of the four Glu120 residues, coordinate a divalent cation. Two independent structures built from MAD-phased data indicated the presence of a strontium and calcium ion bound at this site, respectively. This metal-binding site appears to play an important role in stabilizing the homo-tetramer, which has implications for the engagement of NSP4 as an enterotoxin.

Amino Acid Sequence↗

Acid-base properties of the reaction product of sialic acid with fluorogenic reagent, 1,2-diamino-4,5-methylenedioxybenzene (DMB).

N-Acetylneuraminic acid (Neu5Ac) forms the highly fluorophoric quinoxalinone derivative (Q) when treated with 1,2-diamino-4,5-methylenedioxybenzene (DMB). Effects of protonation and deprotonation on the fluorescence of Q were examined at room temperature. The strong fluorescence was found to be caused by the neutral form Q but not the protonated form of its excited state [Q]* and at pH below 1 the emission was completely quenched. The deprotonated singlet form [Q-]* was a less efficient fluorescer than [Q]*.

Chromatography, High Pressure Liquid↗

Chemical analysis of the developmental pattern of polysialylation in chicken brain. Expression of only an extended form of polysialyl chains during embryogenesis and the presence of disialyl residues in both embryonic and adult chicken brains.

Recent studies have demonstrated the involvement of two polysialyltransferases in neural cell adhesion molecule (N-CAM) polysialylation. The availability of cDNAs encoding these enzymes facilitated studies on polysialylation of N-CAM. However, there is a dearth of detailed structural information on the degree of polymerization (DP), DP ranges, and the influence of embryogenesis on the DP. It is also unclear how many polysialic acid (polySia) chains are attached to a single core N-glycan. In this paper we applied new, efficient, and sensitive high pressure liquid chromatography methods to qualitatively and quantitatively analyze the polySia structures expressed on embryonic and adult chicken brain N-CAM. Our studies resulted in the following new findings. 1) The DP of the polySia chains was invariably 40-50 throughout developmental stages from embryonic day 5 to 21 after fertilization. In contrast, glycopeptides containing polySia with shorter DPs, ranging from 15 to 35, were isolated from adult brain. 2) Chemical evidence showed glycan chains abundant in Neu5Acalpha2,8Neu5Ac were expressed during all developmental stages including adult. 3) Levels of both di- and polySia were found to show distinctive changes during embryonic development.

Animals↗

CROC-4: a novel brain specific transcriptional activator of c-fos expressed from proliferation through to maturation of multiple neuronal cell types.

A novel, brain-specific cDNA, denoted CROC-4, was cloned from human brain by a contingent replication of cDNA procedure capable of detecting transcriptional activators of the human c-fos proto-oncogene promoter. CROC-4 encoded an 18-kDa serine/threonine-rich polypeptide containing a P-loop motif and an SH3-binding region with phosphorylation sites for a variety of protein kinases (cdc2, CDK2, MAPK, CDK5, protein kinase C, Ca(2+)/calmodulin protein kinase 2, casein kinase 2) involved in cell proliferation and differentiation. Immunohistochemistry revealed that during early development, expression was associated with proliferating and migrating cells throughout the rodent brain, initially appearing in the proliferative ventricular zones. During late development and in adult human brain, CROC-4 was expressed in diverse brain regions including the thalamus, subthalamic nucleus, corpus callosum, substantia nigra, caudate nucleus, amygdala, and hippocampus. The association of CROC-4 expression with proliferating regions of developing brain and retention in regions of the adult brain, as well as the punctate nuclear location, suggest that CROC-4 participates in brain-specific c-fos signaling pathways involved in cellular remodeling of brain architecture.

Adult↗

Activation of extracellular signal-regulated kinase (ERK) and Akt by human serotonin 5-HT(1B) receptors in transfected BE(2)-C neuroblastoma cells is inhibited by RGS4.

Regulator of G protein signaling (RGS) proteins are GTPase-activating proteins for heterotrimeric G proteins. One of the best-studied RGS proteins, RGS4, accelerates the rate of GTP hydrolysis by all G(i) and G(q) alpha subunits yet has been shown to exhibit receptor selectivity. Although RGS4 is expressed primarily in brain, its effect on modulating the activity of serotonergic receptors has not yet been reported. In the present study, transfected BE(2)-C human neuroblastoma cells expressing human 5-HT(1B) receptors were used to demonstrate that RGS4 can inhibit the coupling of 5-HT(1B) receptors to cellular signals. Serotonin and sumatriptan were found to stimulate activation of extracellular signal-regulated kinase. This activation was attenuated, but not completely inhibited, by RGS4. Similar inhibition by RGS4 of the protein kinase Akt was also observed. As RGS4 is expressed at high levels in brain, these results suggest that it may play a role in regulating serotonergic pathways.

Calcium-Calmodulin-Dependent Protein Kinases↗

Choledochal cyst associated with acute acalculous cholecystitis.

A 7-year-old girl underwent laparotomy for suspected acute appendicitis. Instead, bile peritonitis with hydrops of gallbladder and normal appendix were noted. Dilatation of the CBD was observed, and choledochal cyst was documented by intraoperative cholangiography. Cholecystectomy and T-tube placement were performed. Postoperative follow-up by ultrasound lasted for 9 months. The CBD remained the same size without clinical manifestation.

Acute Disease↗