Lactate dehydrogenase isoenzyme patterns in auricular pseudocyst fluid.

OBJECTIVE: We investigated lactate dehydrogenase isoenzyme patterns in the cyst fluid of auricular pseudocysts and autogenous blood, to assist the diagnosis of auricular pseudocyst. METHODS: Twenty patients with auricular pseudocysts participated in this study conducted in Kaohsiung Medical University Hospital between February 2007 and June 2010. Patterns of lactate dehydrogenase in cyst fluid and autogenous blood were analysed. RESULTS: Levels of lactate dehydrogenase 1 and 2 were lower in auricular pseudocysts than in autogenous blood, whereas levels of lactate dehydrogenase 4 and 5 were higher; this difference was statistically significant (p < 0.001). CONCLUSION: Lactate dehydrogenase isoenzyme patterns in auricular pseudocyst fluid indicated higher percentage distributions of lactate dehydrogenase 4 and 5 and lower percentage distributions of lactate dehydrogenase 1 and 2. An effective laboratory method of evaluating the different lactate dehydrogenase isoenzyme components was developed; this method may improve the accuracy of auricular pseudocyst diagnosis.

Magnetic resonance venography evaluating veins flow for legs by application of long stretch elastic bandage.

AIM: Varicose veins are a common feature of chronic venous disease that causes pain and swelling of legs. Long stretch elastic bandages are one of the methods used to prevent and treat venous disease, but diagnostic benefit by magnetic resonance imaging (MRI) is not widely promoted. This study aims to evaluate the effects of long stretch elastic bandages using a non-contrast magnetic resonance venographic (MRV) technique by detecting influence on circumferences and flow velocities of varicose and deep veins, before and immediately after application of long stretch elastic bandages. METHODS: Twelve patients presenting with visible varicose veins underwent MRV. The level of the superficial varicose and deep veins before and after wearing long stretch elastic bandages was recorded. RESULTS: By comparing before and after long stretch elastic bandages, all 12 cases showed statistical significant decrease of the superficial venous flow areas; 9 out of 12 cases showed statistical significant change of the superficial (decreased) and deep (increased) venous flow rates. However, due to technical limitation, over half of the cases could not record the change of the deep venous flow areas. CONCLUSION: The non-contrast MRV technique with Cine phase contrast, is capable to detect the short term beneficial effects of long stretch elastic bandages by measuring change of the superficial venous flow areas and superficial and deep venous flow rates.

Abnormal liver function test results are related to metabolic syndrome and BMI in Taiwanese adults without chronic hepatitis B or C.

BACKGROUND: Metabolic syndrome (MS) is considered a cause of abnormal deposition of fat into hepatocytes, which might be associated with hepatic steatosis or abnormal liver function. OBJECTIVE: The aim of this study was to explore the factors associated with MS and the relationship between MS and abnormal aspartate aminotransferase (AST), alanine aminotransferase (ALT) and gamma-glutamyl transferase (GGT) levels in Taiwanese subjects without chronic hepatitis B (CHB) or C (CHC). SUBJECTS: We enrolled 2539 Taiwanese adults without CHB or CHC (age range: 16-88 years old) and investigated the factors related to MS using the NCEP-ATP (National Cholesterol Education Program-Adult Treatment Panel) III criteria; body mass index (BMI) was measured using Asia-Pacific criteria. RESULTS: The prevalence rate of MS in Taiwanese adults without CHB or CHC was 16.9% using the modified ATP III criteria and 15.4% using the International Diabetes Federation criteria. Males had a significantly higher prevalence rate than females (P<0.001), and subjects with MS were significantly older and had significantly higher BMI values and AST, ALT and GGT levels (all P<0.001). In univariate analyses, the abnormality of liver function test results were related to gender, level of fasting sugar, systolic blood pressure, triglyceride, high-density lipoprotein, BMI and MS (all P<0.05). Multivariate analysis showed that the male gender, a higher BMI value and MS were related to abnormal liver function test results. The cutoff value for ALT in relation to MS is 31 IU l(-1) for male and 18 IU l(-1) for female. CONCLUSION: The prevalence of MS in Taiwanese adults without hepatitis B or C was found to be high, and MS and BMI were identified as being related to abnormal liver function test results in these adults.

The endoplasmic reticulum in apoptosis and autophagy: role of the BCL-2 protein family.

Apoptosis is essential for normal development and maintenance of homeostasis, and disruption of apoptotic pathways is associated with multiple disease states, including cancer. Although initially identified as central regulators of apoptosis at the level of mitochondria, an important role for BCL-2 proteins at the endoplasmic reticulum is now well established. Signaling pathways emanating from the endoplasmic reticulum (ER) are involved in apoptosis initiated by stimuli as diverse as ER stress, oncogene expression, death receptor (DR) ligation and oxidative stress, and the BCL-2 family is almost invariably implicated in the regulation of these pathways. This also includes Ca(2+)-mediated cross talk between ER and mitochondria during apoptosis, which contributes to the mitochondrial dynamics that support the core mitochondrial apoptosis pathway. In addition to the regulation of apoptosis, BCL-2 proteins at the ER also regulate autophagy, a survival pathway that limits metabolic stress, genomic instability and tumorigenesis. In cases where apoptosis is inhibited, however, prolonged autophagy can lead to cell death. This review provides an overview of ER-associated apoptotic and autophagic signaling pathways, with particular emphasis on the BCL-2 family proteins.

Risk estimates for drugs suspected of being associated with Stevens-Johnson syndrome and toxic epidermal necrolysis: a case-control study.

The purpose of this case-control study is to estimate the risks of Stevens-Johnson syndrome or toxic epidermal necrolysis associated with the use of specific drugs. The suspected cases were identified from the computerized hospital discharge file. We calculated crude relative risks and adjusted them for confounding by multivariate analysis. The analysis was based on 35 cases and 105 controls. This study showed that the use of carbamazepine, phenytoin and allopurinol is most associated with the risks in the oriental population.

Effect of anti-hypertensive drug dose frequency on the clinic-home blood pressure difference in patients with stage 1 treated hypertension.

Clinic blood pressure (CBP) is generally used for diagnosis and treatment monitoring in hypertension, but target organ damage correlates more closely with home blood pressure (HBP). Eliminating the clinic-home blood pressure difference (CHBPD) would make conventional CBP a more accurate alternative to HBP. This prospective, randomized, open trial compared the effect of a once-daily versus a twice-daily regimen of anti-hypertensive therapy on CHBPD. After a 2-week wash-out period, 85 confirmed stage 1 hypertensive patients were randomized to receive 2 mg trichlormethiazide daily in one (40 subjects) or two (45 subjects) daily doses for 3 weeks. CBP and HBP measurements were taken during the third week of treatment and the CHBPD calculated. After treatment, the systolic and diastolic CHBPD values were significantly greater in the once-daily regimen than in the twice-daily regimen. Conventional CBP should not be used as an alternative to HBP for evaluating prognosis and monitoring anti-hypertensive therapy when using a once-daily regimen.

Restricted expression of LUZP in neural lineage cells: a study in embryonic stem cells.

A novel protein LUZP with 3 leucine zipper motifs at its amino terminus is predominantly expressed in the adult brain. A modified gene targeting approach was employed in an attempt to establish in vitro and in vivo models in which Luzp is knock-out (KO) for phenotype assessment and a reporter gene lacZ is knock-in (KI) for tracing its expression. We report in this study the molecular cloning of the Luzp gene, its targeting vector construction and Luzp-KO/lacZ-KI embryonic stem (ES) clone selection. Since LUZP is also expressed in ES cells, the possibility of embryonic lethality cannot be excluded when attempting to establish Luzp-null mutant mice. We have therefore examined the development of homozygous Luzp-KO/lacZ-KI clones in nude mice. Tissue types derived from all three embryonic germ layers were observed in teratomas developed in nude mice. In situ X-gal staining further revealed restricted expression of LUZP in neural lineage cells.

Computer-aided measurement and grading of cranial asymmetry in children with and without torticollis.

Our aim was to develop a simple, non-invasive method that could be used to objectively record cranial symmetry over time. We utilized this new method to investigate the relationship between torticollis and progressive plagiocephaly. From 1995 to 1999, the head shapes of 419 torticollis patients and 1 211 normal children were evaluated. We used Ezeform strip, a splint material, to make a permanent ring of the head circumference. Each head ring was recorded, scanned, and analyzed. An asymmetric index (AI) was designed. Double-blind comparisons of clinical assessment with AI values demonstrated a good agreement. Asymmetry of the cranium was found with similar frequency in torticollis and normal babies up to 12 months old. After 1 year of age, the deformity persists in patients with torticollis into their adolescence, while the normal group shows increased symmetry. This new recording system offers an objective and efficacious methods for following the natural history of cranial asymmetry.

A macrophage protein, Ym1, transiently expressed during inflammation is a novel mammalian lectin.

Oral infections of mice with Trichinella spiralis induce activation of peritoneal exudate cells to transiently express and secrete a crystallizable protein Ym1. Purification of Ym1 to homogeneity was achieved. It is a single chain polypeptide (45 kDa) with a strong tendency to crystallize at its isoelectric point (pI 5.7). Co-expression of Ym1 with Mac-1 and scavenger receptor pinpoints macrophages as its main producer. Protein microsequencing data provide information required for full-length cDNA cloning from libraries constructed from activated peritoneal exudate cells. A single open reading frame of 398 amino acids with a leader peptide (21 residues) typical of secretory protein was deduced and later deposited in GenBank (accession number M94584) in 1992. By means of surface plasmon resonance analyses, Ym1 has been shown to exhibit binding specificity to saccharides with a free amine group, such as GlcN, GalN, or GlcN polymers, but it failed to bind to other saccharides. The interaction is pH-dependent but Ca2+ and Mg2+ ion-independent. The binding avidity of Ym1 to GlcN oligosaccharides was enhanced by more than 1000-fold due to the clustering effect. Specific binding of Ym1 to heparin suggests that heparin/heparan sulfate may be its physiological ligand in vivo during inflammation and/or tissue remodeling. Although it shares approximately 30% homology with microbial chitinases, no chitinase activity was found associated with Ym1. Genomic Southern blot analyses suggest that Ym1 may represent a member of a novel lectin gene family.

The crystal structure of a novel mammalian lectin, Ym1, suggests a saccharide binding site.

Ym1, a secretory protein synthesized by activated murine peritoneal macrophages, is a novel mammalian lectin with a binding specificity to GlcN. Lectins are responsible for carbohydrate recognition and for mediating cell-cell and cell-extracellular matrix interactions in microbes, plants, and animals. Glycosaminoglycan heparin/heparan sulfate binding ability was also detected in Ym1. We report here the three-dimensional structure of Ym1 at 2.5-A resolution by x-ray crystallography. The crystal structure of Ym1 consists of two globular domains, a beta/alpha triose-phosphate isomerase barrel domain and a small alpha + beta folding domain. A notable electron density of sugar is detected in the Ym1 crystal structure. The saccharide is located inside the triose-phosphate isomerase domain at the COOH terminal end of the beta-strands. Both hydrophilic and hydrophobic interactions are noted in the sugar-binding site in Ym1. Despite the fact that Ym1 is not a chitinase, structurally, Ym1 shares significant homology with chitinase A of Serratia marcescens. Ym1 and chitinase A have a similar carbohydrate binding cleft. This study provides new structure information, which will lead to better understanding of the biological significance of Ym1 and its putative gene members.

Clinical and morphologic features of hypertrophic cardiomyopathy in elderly patients 85 years or older.

We studied the distinctive morphology of the left ventricle (LV) and attempted to relate advanced age and hypertension to this characteristic feature in elderly patients with hypertrophic cardiomyopathy (HC). Fourteen elderly patients > or = 85 years old (mean age 90 +/- 5 years) with HC were compared with 45 young patients < or = 40 years (mean age 34 +/- 4 years) with this disease. More mild hypertension in the elderly (10/14, 71%) than in the young (0%), and more syncope in the young (10/45, 22%) than in the elderly (0%) were observed. Echocardiography showed that the elderly patients had relatively mild LV wall thickening, generally confined to the septum (elderly vs young: 18 +/- 4 vs 25 +/- 8 mm, p < 0.001), with more basal septal bulging (elderly vs young: 12/14, 86% vs 0%, p < 0.001) and anterior septal hypertrophy of LV (elderly vs young: 11/14, 79% vs 0%, p < 0.001). Elderly patients with mild hypertension showed a predominantly basal septal bulging (10/10, 100%) and anterior septal hypertrophy of LV (9/10, 90%). HC in elderly patients > or = 85 years old has a striking LV morphology. Mild hypertension and advanced age may contribute to the distinctive geometry.

alpha-Methylene-gamma-butyrolactones: synthesis and vasorelaxing activity assay of coumarin, naphthalene, and quinoline derivatives.

Certain alpha-methylene-gamma-butyrolactone derivatives of coumarin, naphthalene, and quinoline were synthesized and evaluated for vasorelaxing effects on isolated rat thoracic aorta. The 7-[(2,3,4,5-tetrahydro-2-methyl-4-methylene-5-oxo-2-furanyl)methoxy]-2H- 1- benzopyran-2-ones, which have an aliphatic methyl substituent at the lactone C2, were more active than their C2-phenyl counterparts against high-K+ (80 mM) medium, Ca2+ (1.9 mM)-induced vasoconstriction and the norepinephrine (NE, 3 microM)-induced phasic and tonic constrictions (2a vs. 2b; 2c vs. 2d; 2e vs. 2f; 2g vs. 2h). Although 3-chloro-7-[(2,3,4,5-tetrahydro-2-methyl-4-methylene-5-oxo-2- furanyl)methoxy]-4-methyl-2H-1-benzopyran-2-one (2g) demonstrated the most potent inhibitory activities on the NE-induced phasic and tonic constrictions at concentrations of as low as 10 micrograms/ml, it possesses both affinity for NE-receptor and intrinsic activity to trigger the vasoconstriction. However, 8-[(2,3,4,5-tetrahydro-2-methyl-4-methylene-5-oxo-2- furanyl)methoxy]quinoline (10a) and other quinoline derivatives (11a, 12a) are pure irreversible non-competitive blockers of NE-receptor with no intrinsic activity. The aromatic ring played an important role in the vasorelaxing effects of alpha-methylene-gamma-butyrolactones; naphthalene was inactive, quinolines exhibited only affinity to the alpha-receptor, and coumarins possessed both affinity and intrinsic activity.

Left ventricular filling profiles and angiotensin system activity in elite baseball players.

UNLABELLED: Left ventricular (LV) filling profiles in elite baseball players has not been reported in the literature. Also, angiotensin system activity in athletes has never been reported. We used echocardiography to compare 20 male elite baseball players (aged 21.9+/-1.0 years) with those of age- and sex-matched healthy sedentary subjects. Compared with the normal group, the athlete group showed a significant increase in LV mass, LV diastolic and systolic dimension, and left atrial dimension (P<0.05, <0.001, <0.001, and <0.001, respectively). No differences in relative wall thickness or fractional shortening were found between these two groups. Diastolic filling profiles, including peak early diastolic filling velocity (E), peak late diastolic filling velocity (A), E:A ratio, early time-velocity integral (Ei), atrial time-velocity integral (Ai), Ei:Ai ratio, early filling time, deceleration time of early filling, and isovolumic relaxation time, were similar in both groups. Angiotensin system activity, including plasma renin activity, plasma aldosterone, and 24-h urinary aldosterone excretion, showed no difference between these two groups. CONCLUSION: This study suggests that normal LV filling profile, which is mediated partly by normal angiotensin system activity, is not related to increase in LV dimension and mass in elite baseball players.

Left ventricular filling profiles in young white-coat hypertensive patients without hypertrophy.

This study was to assess left ventricular diastolic function in young white-coat hypertensive subjects < 50 years of age without hypertrophy. Hypertensive patients (systolic or diastolic blood pressure > or = 140 or > or = 90 mm Hg on all three visits) were defined as white coat if their average 24-hour blood pressure was < 127/81 mm Hg and at least 18/16 mm Hg lower than their average office values. We chose three groups balanced for sex, age, and body mass index: 50 sustained hypertensives, 25 white-coat hypertensives, and 25 normotensives. Office blood pressure was similar in white-coat and sustained hypertensives. Ambulatory blood pressure was comparable in white-coat hypertensives and normotensives. Compared with normotensives, white-coat hypertensives had more impaired diastolic function: increased ratio of late to early filling velocities, raised ratio of late to early time-velocity integral, prolonged deceleration time, and lengthened isovolumic relaxation time (P<.001, P<.001, P=.002, and P<.001, respectively). No difference was noticed between white-coat and sustained hypertensives. Compared with normotensives, white-coat hypertensives had higher values of plasma and urine norepinephrine (P<.001 and P<.001, respectively), plasma and urine aldosterone (P<.001 and P=.002, respectively), plasma renin activity (P=.04), total cholesterol (P=.001), and LDL cholesterol (P<.001). No difference was observed between white-coat and sustained hypertensives. Within white-coat hypertensives, 24-hour urinary aldosterone closely correlated with the ratio of late to early filling velocities (P=.008), and plasma and 24-hour urinary norepinephrine correlated well with total cholesterol (P=.037 and P=.006, respectively). No correlation was detected within the sustained hypertensives and normotensives. Heightened neurohumoral activity clearly supported the progression of diastolic dysfunction and metabolic abnormality in white-coat hypertensives.

Differential expression of the GABAA receptor alpha 1 subunit in developing chicken brain.

A unique segment of chicken GABAA receptor alpha 1 subunit was expressed in E. coli and used to generate an antiserum 2A specific for the subunit. The DNA fragment encoding the segment of alpha 1 was obtained by selective amplification by polymerase chain reaction (PCR) from a chicken brain cDNA library. The antiserum is characterized by its capacity to immunoprecipitate a [3H]flunitrazepam binding protein of 50 kDa, the chicken GABAA receptor alpha 1 subunit. Subsequent immunoblotting and immunocytochemistry analyses reveal that alpha 1 is expressed in the optic tectum and cerebellum as early as embryonic day 15 (E15), in various areas of telencephalon as early as E20 and distributed heterogeneously among different cell types. The early expression of alpha 1 may imply its functional significance in neurotransmission.

Myocardial protective effect of trilinolein: an antioxidant isolated from the medicinal plant Panax pseudoginseng.

In a previous study we demonstrated that trilinolein, a natural plant triacylglycerol, is a novel myocardial protective agent in vivo. The mechanism probably involves an antioxidant effect. This work investigated the mechanism of myocardial protection of trilinolein to determine if inhibition of calcium influx and alteration of activity of superoxide dismutase are involved. In isolated cardiomyocytes, pretreatment with trilinolein at a low concentration of 10(-9) M effectively reduced 45Ca2+ influx stimulated by hypoxia/normoxia by 34%. In isolated perfused rat heart subjected to 60 min global hypoxemia without reperfusion, pretreatment with 10(-7) M trilinolein for 15 min reduced infarct size by 37%. Assay of superoxide dismutase-mRNA by Northern blot analysis in in vivo rat heart subjected to 30 min ischaemia and 10 min reperfusion showed pretreatment with 10(-7) M trilinolein had a synergistic action with antioxidant systems preventing the rise in superoxide dismutase-mRNA. These results reconfirm the myocardial protection of trilinolein and suggest it may be related to antioxidant activity and inhibition of 45Ca2+ influx.

Enalapril does not improve left ventricular diastolic dysfunction in young and mild hypertensives without concomitant hypertrophy.

The aim of the study was to determine whether enalapril monotherapy can improve left ventricular diastolic dysfunction (LVDD) in young and mild hypertensive patients without concomitant left ventricular hypertrophy (LVH). Fifty patients with hypertension < or = 160/100 mm Hg, aged < or = 50 years, normal two-dimensional echocardiographic (2-D echo) measurements, and LVDD were enrolled in this study. The LVDD was defined as a transmitral early (E) to atrial (A) peak velocity ratio of < or = 1. The mean documented hypertension was 6.3 years. The mean daily dose of enalapril was 13 mg. Baseline and 24-month follow-up echocardiograms were evaluated. Thirty-eight age- and sex-matched healthy subjects served to establish the normal reference values of 2-D echo measurements. After treatment, peak early diastolic velocity (E) (49 +/- 6 cm/sec v 48 +/- 10 cm/sec; P = not significant), peak atrial velocity (A) (62 +/- 9 cm/sec v 62 +/- 10 cm/sec; P = not significant), and E/A ratio (0.80 +/- 0.10 v 0.78 +/- 0.13; P = not significant) remained unchanged. Moreover, early to atrial velocity-time integral ratio (1.24 +/- 0.08 v 1.23 +/- 0.11; P = not significant) did not change. The left ventricular mass index, relative wall thickness, left ventricular end-systolic diameter, left atrial diameter, fractional shortening, heart rate, and body mass index did not show significant changes in all hypertensive patients. In conclusion, long-term antihypertensive therapy with enalapril did not lead to an improvement of LVDD in young and mild hypertension patients without concomitant LVH.

Identification, molecular characterization, and chromosomal localization of the cDNA encoding a novel leucine zipper motif-containing protein.

cDNA clones encoding a novel protein (LUZP) with three leucine zipper motifs were first identified from a murine bone marrow cDNA library. After screening two additional cDNA libraries of activated peritoneal exudate cells, 32 positive clones were obtained from 1.3 x 10(7) phage plaques. Four overlapping clones constituting a total of 7399 bp were sequenced on both strands. The complete open reading frame of LUZP is 1067 amino acids. In addition to three leucine zipper motifs located at the NH2 terminus, there are three nuclear localization signals and a large number of putative Ser/Thr phosphorylation sites. Western blot analyses indicate that LUZP is predominantly expressed in brain, whereas immunocytochemistry data clearly reveal its presence in the nucleus of neurons. Interspecific backcross analyses have mapped Luzp to mouse chromosome 4 in proximity to Gpcr14. Comparative mapping data suggest that the human homolog of Luzp will map to human chromosome 1p36.

Characterization of the regulatory regions of murine alpha 2C2 adrenoceptor subtype gene.

In order to delineate the regulatory mechanisms underlying the control of alpha 2 adrenoceptor expression, the sequence of 5' (1145 bp) and 3' (2682 bp) flanking regions of murine alpha 2C2 subtype gene were determined and characterized from a genomic phage clone MA2C2. The 5' flanking region has no TATA box yet with high GC content. The 3' flanking region is marked by the presence of a polyadenylation signal 2.3 kb down stream from the stop codon. The transcription start site was mapped by 5' rapid amplification of cDNA ends (RACE) and primer extension assays at nucleotide A, 415 bp upstream from the first initiation codon and resides in a motif resembling the consensus sequence of initiator found in many TATA-less promoters. An NcoI fragment (4.7 kb) immediately upstream from the translation initiation site was linked to a reporter gene lacZ. Using an in vitro transfection assay, cell lines of renal or neural origin were identified as permissive hosts for alpha 2C2 subtype expression. With its core promoter clearly defined and sequence of the regulatory regions at hand, this in vitro gene transfer system will facilitate the identification of putative cis-elements and transcription factors key to alpha 2C2 adrenoceptor expression.