Visceral adipose tissue, but not waist circumference is a better measure of metabolic risk in Singaporean Chinese and Indian men.

OBJECTIVES: Visceral adipose tissue (VAT) is an independent risk factor in cardiometabolic diseases and is commonly measured by computed tomography (CT). It is measured clinically by waist circumference (WC). The L4/5 intervertebral space VAT (L4/5 VAT) is traditionally used to represent total VAT volume. We set out to determine (1) the level of intervertebral space on CT that best approximates the total VAT volume; (2) compare the association between WC and VAT in Singaporean Chinese and Indian; and (3) examine the correlation between VAT and cardiometabolic risk factors. SUBJECTS: A total of 60 Chinese and 60 Asian Indian men older than 60 years were recruited. Their medical history was taken and anthropometry was measured. Fasting glucose, insulin, lipids, adipokines and inflammatory markers were measured. Insulin resistance was evaluated by homeostasis model assessment-insulin resistance. VAT was determined by CT. Total VAT volume was calculated in 22 patients from VAT areas at seven intervertebral levels. The optimal VAT area most representative of total VAT volume was determined and used for all patients to approximate total VAT volume. RESULTS: The VAT area at L2/3 intervertebral space (L2/3 VAT) correlated almost perfectly with VAT volume (R(2)=0.974 and 0.946 for Chinese and Indians, respectively). SUBJECTS from the two races had similar height, weight, body mass index (BMI), WC and L2/3 VAT but more Indian men had hypertension, hyperlipidemia and type 2 diabetes mellitus. WC was correlated with the L2/3 VAT area in both Chinese (r=0.484, P<0.001) and Indian subjects (r=0.366, P=0.004) without racial difference (P=0.2 for interaction term). L2/3 VAT also correlated better with cardiometabolic risk factors, adipokines and C-reactive protein than WC, BMI or L4/5 VAT. CONCLUSION: The L2-L3 intervertebral space was the best anatomic level for a single-slice CT cross-sectional area measurement of VAT to approximate total body visceral adipose volume in this population of Chinese and Asian Indian men older than 60 years. L2/3 VAT was better correlated with multiple cardiovascular risk factors, adipokines and inflammatory marker than either L4/5 VAT, WC or BMI.

Micromechanical testing with microstrain resolution.

Simple test equipment has been developed for studying the elastic limit and plastic deformation of thin metal wires and thin foils (down to 10 µm) under torsion, tension, and bending. Using load-unload methods and gauge lengths up to 1 m, plastic strain as low as 10(-6) can be measured accurately.

A novel isoform of the 8p22 tumor suppressor gene DLC1 suppresses tumor growth and is frequently silenced in multiple common tumors.

The critical 8p22 tumor suppressor deleted in liver cancer 1 (DLC1) is frequently inactivated by aberrant CpG methylation and/or genetic deletion and implicated in tumorigeneses of multiple tumor types. Here, we report the identification and characterization of its new isoform, DLC1 isoform 4 (DLC1-i4). This novel isoform encodes an 1125-aa (amino acid) protein with distinct N-terminus as compared with other known DLC1 isoforms. Similar to other isoforms, DLC1-i4 is expressed ubiquitously in normal tissues and immortalized normal epithelial cells, suggesting a role as a major DLC1 transcript. However, differential expression of the four DLC1 isoforms is found in tumor cell lines: Isoform 1 (longest) and 3 (short thus probably nonfunctional) share a promoter and are silenced in almost all cancer and immortalized cell lines, whereas isoform 2 and 4 utilize different promoters and are frequently downregulated. DLC1-i4 is significantly downregulated in multiple carcinoma cell lines, including 2/4 nasopharyngeal, 8/16 (50%) esophageal, 4/16 (25%) gastric, 6/9 (67%) breast, 3/4 colorectal, 4/4 cervical and 2/8(25%) lung carcinoma cell lines. The functional DLC1-i4 promoter is within a CpG island and is activated by wild-type p53. CpG methylation of the DLC1-i4 promoter is associated with its silencing in tumor cells and was detected in 38-100% of multiple primary tumors. Treatment with 5-aza-2'-deoxycytidine or genetic double knockout of DNMT1 and DNMT3B led to demethylation of the promoter and reactivation of its expression, indicating a predominantly epigenetic mechanism of silencing. Ectopic expression of DLC1-i4 in silenced tumor cells strongly inhibited their growth and colony formation. Thus, we identified a new isoform of DLC1 with tumor suppressive function. The differential expression of various DLC1 isoforms suggests interplay in modulating the complex activities of DLC1 during carcinogenesis.

Elastic limit and strain hardening of thin wires in torsion.

A theory for the size effect in the strength of wires under torsion is reported and compared with data from thin copper wires. Critical thickness theory is solved rigorously and used to validate a useful approximation which is combined with slip-distance theory modified for a finite structure size. Experimental data with high accuracy around and above the elastic limit show excellent agreement with the theory. The results strongly imply that the physical principle is the constraint that size, whether grain size or structure size, puts on allowed dislocation curvature.

Three-dimensional MR volumetric analysis of the posterior fossa CSF space in hemifacial spasm.

BACKGROUND: We hypothesize that a smaller posterior fossa (PF) CSF space may be a risk factor for hemifacial spasm (HFS). OBJECTIVE: We conducted a case-control 3-dimensional magnetic resonance (MR) volumetric study in patients with HFS and determined the clinical predictive factors of PF CSF volume. METHODS: Patients with clinically diagnosed HFS and controls matched for age, sex, race, and hypertension underwent MRI/magnetic resonance angiography examination. The PF CSF space was segmented and quantified on a heavily T2-weighted high-resolution 3-dimensional MR volume slab, centered over the porus acusticus. RESULTS: Eighty-two study subjects (41 patients and 41 controls) were included. The mean PF CSF volume in patients with HFS and controls was 17,303.0 +/- 3,900.0 vs 19,216.0 +/- 3,912.0 mm(3). The mean volume in patients with HFS was 11.4% smaller than in controls (p = 0.015). Analysis of differences between individually matched pairs and controls also revealed that PF CSF for controls was larger than that for patients with HFS (p = 0.007). A multivariate linear regression analysis revealed that a small PF CSF volume was associated with HFS (p = 0.01). Decreasing age (p = 0.001) and female gender (p < 0.0005), but not hypertension (p = 0.892), were also found to be predictors of a low PF CSF volume. CONCLUSIONS: Our results showed that the posterior fossa (PF) CSF volume was lower in patients with HFS compared with matched controls. HFS, female gender, and younger age were associated with smaller PF CSF volume. These observations could explain the strong female preponderance in both clinic- and population-based epidemiologic studies.

Physio-chemical reactions in recycle aggregate concrete.

Concrete waste constitutes the major proportion of construction waste at about 50% of the total waste generated. An effective way to reduce concrete waste is to reuse it as recycled aggregate (RA) for the production of recycled aggregate concrete (RAC). This paper studies the physio-chemical reactions of cement paste around aggregate for normal aggregate concrete (NAC) and RAC mixed with normal mixing approach (NMA) and two-stage mixing approach (TSMA) by differential scanning calorimetry (DSC) and scanning electron microscopy (SEM). Four kinds of physio-chemical reactions have been recorded from the concrete samples, including the dehydration of C(3)S(2)H(3), iron-substituted ettringite, dehydroxylation of CH and development of C(6)S(3)H at about 90 degrees C, 135 degrees C, 441 degrees C and 570 degrees C, respectively. From the DSC results, it is confirmed that the concrete samples with RA substitution have generated less amount of strength enhancement chemical products when compared to those without RA substitution. However, the results from the TSMA are found improving the RAC quality. The pre-mix procedure of the TSMA can effectively develop some strength enhancing chemical products including, C(3)S(2)H(3), ettringite, CH and C(6)S(3)H, which shows that RAC made from the TSMA can improve the hydration processes.

Adjuvant lipiodol I-131 after curative resection/ablation of hepatocellular carcinoma.

AIM: A total of 329 patients with hepatocellular carcinoma have been treated at our unit since 1990. Following the randomized controlled trial in Hong Kong by Lau et al. in 1999, patients have been offered adjuvant lipiodol I-131. The aim of this study was to determine the effectiveness of adjuvant lipiodol I-131, following potentially curative surgery with resection and/or ablation, on overall and disease-free survival rates. MATERIAL AND METHODS: The prospectively updated hepatocellular carcinoma database was analysed retrospectively. A total of 34 patients were identified to have received adjuvant lipiodol I-131 post-curative treatment with surgical resection and/or ablation. Patient demographics, clinical, surgical, pathology, and survival data were collected and analysed. RESULTS: Three patients received ablation alone, 24 resection, and 7 resection and ablation. Of the 34 patients treated, there were 2 possible cases of treatment-related fatality (pneumonitis and liver failure). Potential prognostic factors studied for effect on survival included age, gender, serum AFP concentration, Child-Pugh score, cirrhosis, tumor size, portal vein tumor thrombus, tumor rupture, and vascular and margin involvement. The median follow-up duration was 23.3 months. The overall median survival was 40.1 months, while the overall survival rates at 1, 2, 3, and 4 years were 87.1%, 71.7%, 60.7%, and 49.6%, respectively. Median duration to recurrence was 22.3 months. CONCLUSION: Administration of adjuvant lipiodol I-131 is associated with good overall survival.

Epigenetic disruption of interferon-gamma response through silencing the tumor suppressor interferon regulatory factor 8 in nasopharyngeal, esophageal and multiple other carcinomas.

16q24 is frequently deleted in multiple tumors including cancers of nasopharynx, esophagus, breast, prostate and liver. By array comparative genomic hybridization (aCGH), we refined a 16q24 hemizygous deletion in nasopharyngeal carcinoma (NPC) cell lines. Semi-quantitative RT-PCR analysis revealed interferon regulatory factor 8 (IRF8) as the only downregulated gene within this deletion. IRF8 belongs to a family of interferon (IFN) regulatory factors that modulate various important physiologic processes including host defense, cell growth and differentiation and immune regulation. In contrast to the broad expression of IRF8 in normal adult and fetal tissues, transcriptional silencing and promoter methylation of IRF8 were frequently detected in multiple carcinoma (except for hepatocellular) cell lines (100% in NPC, 88% in esophageal and 18-78% in other carcinoma cell lines) and in a large collection of primary carcinomas (78% in NPC, 36-71% in other carcinomas). Methylation of the IRF8 promoter led to the disruption of its response to IFN-gamma stimulation. Pharmacological and genetic demethylation could restore IRF8 expression, indicating a direct epigenetic mechanism. Ectopic expression of IRF8 in tumor cells lacking its expression strongly inhibited their clonogenicity, confirming its tumor suppressor function. Thus, IRF8 was identified as a functional tumor suppressor, which is frequently silenced by epigenetic mechanism in multiple carcinomas.

Intron 2 (T/C) CYP46 polymorphism is associated with Alzheimer's disease in Chinese patients.

BACKGROUND: Cholesterol metabolism has been implicated in the pathophysiology of Alzheimer's disease (AD), and cholesterol-related genes are plausible candidate genes for AD. Genetic association of CYP46A1 polymorphisms with AD had been under extensive investigations; however, observations on intron 2 T-->C (rs754203) generated inconclusive results. OBJECTIVE: To analyse an independent data set in a Chinese population to see whether the polymorphic site rs754203 of the CYP46A1 gene is associated with AD. METHODS: We analysed 130 sporadic AD patients and 110 healthy controls of the Southern Chinese origin. RESULTS: An association between the genotype frequency and AD was suggested in the general population (p = 0.047, odds ratio, OR = 1. 61, 95% confidence interval, CI = 0.96-2.70), while the association was most significant in the apolipoprotein E (ApoE) epsilon4-negative group (p = 0.004, OR = 2.54, 95% CI =1.31-4.95). Linkage disequilibrium block prediction results also favoured this association. Consistent with previous reports, intron 3 C-->T (rs4900442) polymorphism did not show any evidence of association; in our data set ApoEepsilon4 was confirmed to be a genetic risk factor for AD (p = 0.0016, OR = 2.76, 95% CI = 1.50-5.11).

Modulation of invasive properties of murine squamous carcinoma cells by heterologous expression of cathepsin B and cystatin C.

Murine SCC-VII squamous carcinoma cells have the capacity to penetrate reconstituted basement membranes (Matrigel) in vitro. The invasion of Matrigel layers by SCC-VII cells was significantly reduced by E-64, a specific inhibitor of lysosomal cysteine proteinases. The cathepsin-B-selective E-64 derivative, CA-074, inhibited penetration of Matrigel by SCC-VII cells to the same extent, indicating a major role for this particular lysosomal enzyme in extracellular-matrix degradation during squamous-carcinoma-cell invasion. SCC-VII cells were stably transfected with a cDNA encoding human procathepsin B, in an attempt to modulate the invasive properties of the cell line. The transfected cells expressed the heterologous gene, secreted increased amounts of procathepsin B and displayed enhanced invasive potential. In vivo, the activity of cathepsin B is strictly regulated by endogenous inhibitors. SCC-VII cells were therefore also stably transfected with a cDNA encoding human cystatin C, the most potent cysteine-proteinase inhibitor in mammalian tissues. The expression of this transgene resulted in the production of active recombinant cystatin C and a pronounced reduction in Matrigel invasion. These studies demonstrate that the invasive properties of squamous-cell carcinomas can be changed by modulation of the balance between cathepsin B and its endogenous inhibitors, and provide further evidence for the involvement of this lysosomal cysteine proteinase in tumour invasion and metastasis.

Leukaemia inhibitory factor is expressed by normal human keratinocytes in vitro and in vivo.

Leukaemia inhibitory factor (LIF) is a pleotropic cytokine, regulating differentiation, cell growth, cachexia and inflammation. Using the reverse transcription-polymerase chain reaction (RT-PCR) we found that, in culture, normal human keratinocytes (KC) expressed mRNA transcripts for both LIF and the LIF receptor. In the conditioned medium (CM), constitutive LIF protein production was barely detectable but stimulation of KC with 10 ng/ml of either interleukin (IL)-1 alpha, or IL-8, for 24 h, resulted in small but significant increases (P < 0.05) in LIF protein, as measured by enzyme-linked immunosorbent assay. After culture in media containing 1.5 mmol/l calcium, a time-dependent increase in LIF mRNA was seen up to 72 h (an 8.5-fold increase), over levels in cells cultured in 0.05 mmol/l calcium. A large increase in LIF protein in the CM (from 1.15 +/- 0.15 pg/ml to 178.7 +/- 75.7 pg/ml) was seen 72 h after a switch to media containing 1.5 mmol/l calcium (P = 0.05). Twenty-four hours after stimulation of human KC in culture with 10 ng/ml recombinant LIF, a twofold increase in both IL-1 alpha and IL-8 protein in the CM (P < 0.05) was observed. In normal human scalp and foreskin, the epidermis was shown to contain LIF protein by immunostaining. LIF staining was found throughout the epidermis, and in the cells of the outer layer of the root sheath. Thus, KC synthesize LIF in vitro and in vivo.

In vitro and in vivo percutaneous absorption of 14C-chloroform in humans.

Chloroform has been found in potable water and there is concern that significant dermal absorption may arise from daily bathing and other activities. The present study examines percutaneous absorption of 14C-chloroform in vivo using human volunteers and in vitro using fresh, excised human skin in a flow-through diffusion cell system. Fifty microlitre doses of either 1000 micrograms ml-1 chloroform in distilled water, (16.1 micrograms cm-2) or 5000 micrograms ml-1 of chloroform in ethanol, (80.6 micrograms cm-1) were applied to the forearm of volunteers with exhaled air and urine being collected for analysis. Single doses of either 0.4 microgram ml-1 chloroform in distilled water (low dose, 0.62 microgram cm-2, 1.0 ml dosed) or 900 micrograms ml-1 chloroform in distilled water (high dose, 70.3 micrograms cm-2, 50 microliters dosed) were applied to discs of the excised abdominal skin placed in flow-through diffusion cells and perfused with Hepes buffered Hank's balanced salt solution, with a wash at 4 h. In vivo absorption was 7.8 +/- 1.4% (water as vehicle) and 1.6 +/- 0.3% (ethanol as vehicle). Of the dose absorbed in vivo, more than 95% was excreted via the lungs (over 88% of which was CO2), and the maximum pulmonary excretion occurred between 15 min and 2 h after dosing. The percentage of dose absorbed in vitro (skin+perfusate) was 5.6 +/- 2.7% (low dose) and 7.1 +/- 1.4% (high dose). The above data demonstrate that a significant amount of the dissolved chloroform penetrates through the human skin, and that a higher percentage of the applied dose was absorbed using water as vehicle.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparative metabolism of phenanthrene in the rat and guinea pig.

In the present study the biotransformation of phenanthrene in the rat and guinea pig was investigated. 14C-labelled phenanthrene was administered by gavage in corn oil to Sprague-Dawley rats (10 mg/kg b.w./day) and guinea pigs (10 mg/kg b.w./day). Urine and feces were separately collected for the determination of the radioactivity content, and pooled urine was used for the analysis of metabolites. Phenanthrene was metabolized by the rat and guinea pig to free hydroxylated phenanthrenes and their conjugates. The percentages of conjugates, expressed as the total urinary radioactivity, were 39% glucuronides, 24% sulfates and 18% cysteinylglycine for rats; and 39% glucuronides, 23% sulfates and 28% cysteinylglycine for guinea pigs. Enzymatic hydrolysis of glucuronides and sulfates resulted in the formation of free 1,2-, 3,4- and 9,10-dihydrodiols of phenanthrene and 1-, 2-, 3-, and 4-hydroxyphenanthrene in both species.

Percutaneous absorption and metabolism of pyrene, benzo[a]pyrene, and di(2-ethylhexyl) phthalate: comparison of in vitro and in vivo results in the hairless guinea pig.

The in vitro and in vivo absorption and metabolism of pyrene, benzo[a]pyrene, and di(2-ethylhexyl) phthalate (DEHP) were investigated in the hairless guinea pig. The in vitro method, which involved the use of flow-through diffusion cells and Hepes-buffered Hanks' balanced salt solution containing 4% bovine serum albumin as perfusate, was demonstrated to be a suitable system for predicting in vivo absorption of the above lipophilic compounds. The successful application of the in vitro technique for these compounds is significant because no satisfactory in vitro method has hitherto been developed to predict in vivo absorption of highly lipophilic chemicals. Quantification of parent compounds and metabolites that permeated into perfusates and those that remained in skin discs provided insight into the process by which the chemicals penetrated through the skin. Pyrene was absorbed primarily by a passive diffusion process, although a small fraction of the administered dose was biotransformed into metabolites in the skin and partitioned into the receptor fluid. Absorption of benzo[a]pyrene was mediated by biotransformation processes. A metabolite derived from the ultimate carcinogen of this compound, benzo[a]pyrene r-7, t-8,9,10-tetrahydrotetrol, was identified in the receptor fluid. Most of the administered DEHP remained in the skin and only a very small fraction of the dose partitioned into the receptor fluid in either viable or nonviable skin. Data from the present study led to the conclusion that the in vitro method can be utilized to predict in vivo absorption for compounds of high lipophilicity and that dermal metabolism facilitates partitioning of metabolites into the receptor fluid and hence may affect the biological activities of dermally applied compounds.

Percutaneous absorption/metabolism of phenanthrene in the hairless guinea pig: comparison of in vitro and in vivo results.

The in vitro and in vivo percutaneous absorption/metabolism of phenanthrene was investigated in hairless guinea pigs. Flow-through diffusion cells and Hepes-buffered Hanks' balanced salt solution (HHBSS) as receptor fluid were used in the in vitro system. When phenanthrene was applied to excised guinea pig skin mounted on the cells at dose levels of 6.6 and 15.2 micrograms/cm2, 89.7 and 79.1% of the administered doses were respectively absorbed into the skin and receptor fluids during a 24-hr perfusion period. These results are consistent with the in vivo data which showed approximately 80% absorption over the same period of time. Phenanthrene was metabolized in vitro into phenanthrene 9,10-dihydrodiol, 3,4-dihydrodiol, 1,2-dihydrodiol, and traces of hydroxy phenanthrenes. Of the materials absorbed in vitro, 92% was the parent compound and 7% the dihydrodiol metabolites. When a nonviable in vitro system was used, 68% of the applied 15.2 micrograms/cm2 dose was absorbed. Data from the present study demonstrate that the in vitro system is a good model for predicting in vivo percutaneous absorption of phenanthrene, and that penetration of phenanthrene through the skin is controlled more by the passive rate of diffusion than by metabolism.

Replacement of the 3'-CH group by nitrogen in the carbocyclic analogue of thymidine.

We have prepared (4R)-4-thyminyl-D-prolinol, an analogue of 3'-deoxythymidine in which the sugar has been replaced by D-prolinol. This strongly basic secondary amine has been converted to the corresponding hydroxylamine, an analogue of either thymidine or 2'-deoxyxylofuranosylthymine. We have also synthesized a number of simple derivatives of the amine for testing in vitro activity against herpes simplex 1 (HSV-1), human immunodeficiency virus 1 (HIV-1), and a panel of human tumor cell lines. Among these compounds, the hydroxylamine 12 proved active against the human tumor cell lines of breast, colon, and lung origin, with IC50 values of 0.08, 14.02, and 6.91 microM, respectively.