Impact of blast shock waves (SW) with the body wall produces blast lung injuries characterized by bilateral traumatic hemorrhages. Such injuries often have no external signs, are difficult to diagnose, and therefore, are frequently underestimated. Predictive assessment of acute respiratory distress syndrome outcome in SW-related accidents should be based on experimental data from appropriate animal models. Blood plasma transferrin is a major carrier of blood iron essential for proliferative "emergency" response of hematopoietic and immune systems as well as injured tissue in major trauma. Iron-transferrin complexes (Fe3+ TRF) can be quantitatively analyzed in blood and tissue samples with low-temperature EPR techniques. We hypothesized that use of EPR techniques in combination with assays for pro-inflammatory cytokines and granulocytes in the peripheral blood and BAL would reveal a pattern of systemic sequestration of (Fe3+)TRF that could be useful for development of biomarkers of the systemic inflammatory response to lung injury. With this goal we (i) analyzed time-dependent dynamics of (Fe3+)TRF in the peripheral blood of rats after impacts of SW generated in a laboratory shock-tube and (ii) assayed the fluctuation of granulocyte (PMN) counts and expression of CD11b adhesion molecules on the surface of PMNs during the first 24 h after SW induced injury. Sham-treated animals were used as control. Exposure to SW led to a significant decrease in the amount of blood (Fe3+)TRF that correlated with the extent of lung injury and developed gradually during the first 24 h. Thus, sequestration of (Fe3+)TRF occurred as early as 3 h post-exposure. At that time, the steady state concentration of (Fe3+)TRF in blood samples decreased from 19.7+/-0.6 microM in controls to 7.5+/-1.3 microM in exposed animals. The levels of (Fe3+)TRF remained decreased throughout the entire study period. PMN counts increased 5-fold and 3.5-fold over controls respectively, at 3 and 6 h postexposure. These effects were accompanied by an increase in expression of CD11b on the surface membrane of PMNs. Extensive release of cytokines IL-1, IL-6, MCP-1, and MIP-2 was observed in BAL fluid and blood plasma during 24 h postexposure. We conclude that EPR monitoring of blood (Fe3+)TRF can be a useful approach for assessment of systemic pro-inflammatory alterations due to SW-induced lung injury.

BACKGROUND

Leukocyte telomere length, a putative marker of ageing, is a highly variable and heritable complex trait. In order to determine the possible underlying genetic variants for leukocyte telomere length variation, we conducted an association study of leukocyte telomere length and two candidate genes for ageing-related traits, TGFB1 and KLOTHO, in a female Caucasian dizygotic twin population.

METHODS

Terminal restriction fragment (TRF) length, an index of telomere length, was measured using Southern Blotting. Six and four single nucleotide polymorphisms (SNP) were genotyped in TGFB1 and KLOTHO gene, respectively, and tested for association. When there is strong LD between SNPs (r(2)>> 0.5), haplotypic association was investigated using haplotype trend regression approach.

RESULTS

All SNPs were in Hardy-Weinberg equilibrium (p>> 0.05). No significant association was detected for individual SNPs (p>> 0.101), or two-locus haplotypes (p = 0.7497) with TRF variation.

CONCLUSIONS

We failed to find any significant association between leukocyte telomere length and 10 SNPs in two ageing-related candidate genes, TGFB1 and KLOTHO. This result suggests that while we could not exclude minor effects, none of 10 SNPs in these two candidate genes showed significant association with the variation of leukocyte telomere length in our cohort. But as it is unclear whether telomere length dynamics is the cause or the effect of the ageing process, it is still possible the genes are associated with ageing via alternate mechanisms.

In the field of cell-based therapy and regenerative medicine, clinical application is the ultimate goal. However, one major concern is: does in vitro manipulation during culture expansion increases tumourigenicity risk on the prepared cells? Therefore, the aim of this study was to investigate the effect of long-term in vitro expansion on human adipose-derived stem cells (ASCs). The ASCs were harvested from lipo-aspirate samples and cultured until passage 20 (P20), using standard culture procedures. ASCs at P5, P10, P15 and P20 were analysed for morphological changes, DNA damage (Comet assay), tumour suppressor gene expression level (quantitative PCR), p53 mutation, telomerase activity, telomere length determination and in vivo tumourigenicity test. Our data showed that ASCs lost their fibroblastic feature in long-term culture. The population doubling time of ASCs increased with long-term culture especially at P15 and P20. There was an increase in DNA damage at later passages (P15 and P20). No significant changes were observed in both p53 and p21 genes expression throughout the long-term culture. There was also no p53 mutation detected and no significant changes were recorded in the relative telomerase activity (RTA) and mean telomere length (TRF) in ASCs at all passages. In vivo implantation of ASCs at P15 and P20 into the nude mice did not result in tumour formation after 4 months. The data showed that ASCs have low risk of tumourigenicity up to P20, with a total population doubling of 42 times. This indicates that adipose tissue should be a safe source of stem cells for cell-based therapy.

OBJECTIVE

Telomerase is a ribonucleoprotein that synthesizes telomeric DNA. Immortalized and carcinoma cells show no loss of telomere length during cell division. Telomerase activity has been demonstrated in carcinomas of various organs, but not in nonneoplastic tissues. In patients with esophageal carcinoma, no data have been reported concerning the relationship between telomerase activity and clinicopathological findings.

METHODS

Esophageal carcinomas from 31 patients and normal esophageal mucosae from 92 patients were examined. Telomeric Repeat Amplification Protocol assay to detect telomerase activity and Southern blot analysis to examine telomere length were performed.

RESULTS

Of the 31 carcinomas, 27 (87%) had detectable telomerase activity. Twenty-one (23%) of the 92 normal esophageal mucosae from autopsied patients also had detectable telomerase activity. There was no difference between stage and outcome and absence or presence of telomerase activity. No difference in terminal restriction fragment (TRF) length was observed between carcinomas with and without telomerase activity.

CONCLUSIONS

Telomerase activity was demonstrated in a considerable number of normal esophageal mucosae. This suggests the possibility of a high frequency of false positivity if the presence of telomerase activity alone is used as a tumor-specific marker.

Administration of cyproheptadine for four months to a patient with Nelson's Syndrome was associated with evidence of skin lightening, decreased plasma ACTH concentrations, return of a normal circadian periodicity of plasma ACTH concentrations, and lack of responsiveness of plasma ACTH concentrations to TRF administration, in contrast to the response seen in the untreated state.

Human immunodeficiency virus type 1 (HIV-1) infection is accompanied by peripheral CD4+ T-cell losses. CD4+ T-cell numbers often increase during antiviral treatment of acquired immune deficiency syndrome (AIDS), however, alterations in the CD4+ T-cell repertoire have not been completely corrected for these patients. Such individuals remain at increased risk of infection. Although senescence of the CD4+ T cells has not been adequately evaluated for advanced HIV-1 infection, hypothetically, replicative senescence could complicate therapeutic reconstitution of the CD4+ T cells in AIDS. In this study, correlates of replicative senescence, terminal restriction fragment (TRF) length and percentage short (< 5.0 kb) telomeric DNA (senescence fraction), were measured for the CD4+ T cells of HIV-1-infected patients with peripheral CD4+ T-cell counts of < 200/mm3. The results show that for advanced HIV-1 infection the TRF length of the CD4+ T cells is decreased (P < 0.01), and the senescence fraction increased (P < 0.05), when compared with uninfected controls. These findings suggest that cellular senescence may contribute to disruption of CD4+ T-cell diversity observed following the therapeutic, immunologic reconstitution of AIDS.

Fenitrothion (FNT) is an organophosphate compound widely used as pesticide in Malaysia. The present study aims to investigate effects of palm oil tocotrienol rich fraction (TRF) on the renal damage of FNT-treated rats. A total of 40 male Sprague Dawley rats were divided into 4 groups randomly, the control, TRF, FNT and FNT+TRF groups. FNT (20 mg/kg b.w.) and TRF (200 mg/kg b.w.) were given orally for 28 days continuously. Rats from the FNT+TRF group were supplemented with TRF 30 minutes prior to administration of FNT. Rats were sacrificed after 28 days, and the kidneys were removed for determination of oxidative stress and histological analysis. Plasma was collected for determination of blood creatinine and urea level. Statistical analysis showed that palm oil TRF has a protective effect against renal oxidative damage induced by FNT. In the FNT+TRF group, malondialdehyde and protein carbonyl levels were significantly lower, while the glutathione level as well as superoxide dismutase and catalase activities were significantly higher compared with the FNT-treated group (p<0.05). As for renal function, there was a markedly lower urea level (p<0.05) in the FNT+TRF group compared with the FNT-treated group, but there was no significant difference in creatinine level. Besides, total protein also showed no significant difference for all groups of rats (p>0.05). Histological evaluation also revealed that the FNT+TRF group had less glomerulus and renal tubule damage than the FNT-treated group. In conclusion, palm oil TRF was able to reduce oxidative stress and renal damage in FNT-treated rats.

Chimeric genes containing either the mouse transferrin (Trf) enhancer/promoter fused to the structural sequences encoding bovine growth hormone (GH) or the mouse albumin (Alb) enhancer/promoter fused to the gene for human growth hormone-releasing factor (GRF) were microinjected into sheep zygotes. A low percentage of resulting transgenic sheep chronically expressed the respective genes, resulting in elevated plasma concentrations of circulating GH or GRF, respectively. Growth hormone-releasing factor expression induced elevated plasma levels of endogenous GH production. In addition, elevated levels of circulating insulin-like growth factor-I were observed in the bovine GH-expressing Trf transgenic sheep. Growth of these founder transgenic sheep relative to controls were not enhanced. In part, this may be due to the development of the diabetic condition exhibited by both transgenic groups. These results demonstrate that the mouse Trf and Alb enhancer/promoters are active in sheep and suggest that alternate strategies for expressing growth-related genes may be required to modulate growth in sheep.

Receptors for human transferrin (Trf) in high density are found on reticulocytes and syncytiotrophoblast, but most unstimulated, normal adult cells do not bind Trf. In contrast, leukemia and breast adenocarcinoma cells have been shown to manifest Trf receptors, raising the possibility that these receptors might be employed to bind cytotoxic Trf conjugates. Trf was conjugated with adriamycin (Adr) and it was shown that the conjugates are bound by Trf receptors on plasma membranes of Daudi and HL-60 cells, following which Adr is identified in the nuclei of these cells. The biological effect of Adr is quantitated by the inhibition of tritiated thymidine uptake, and subsequent cell death is measured by trypan blue exclusion. The killing correlates directly with both the time of exposure and the amount of conjugate employed. These results suggest that such cytotoxic Trf conjugates hold promise for selective in vivo killing of some malignant cells.

In this study, we investigated the levels of serum Zn, Cu, Mg, Mn, Fe, ceruloplasmin (Cp), transferrin (Trf), and albumin in laryngeal carcinoma and correlated their levels with the cancer stage. The sera from 35 patients with laryngeal cancer (10 at stage II, 12 at stage III, and 13 at stage IV) were extracted before treatment and compared with those from the healthy control group (n=15). Although serum Fe and Mn concentrations were lower in the laryngeal cancer groups (for all stages) than in the control group, the difference was not statistically significant (p>0.05). The higher Cu (p<0.001) and Cp (p<0.01) and lower Zn (p<0.01), Mg (p<0.001), and Trf (p<0.01) concentrations were found in laryngeal cancer groups (for each stage) when compared to the control group. In the comparison of stages II, III, and IV (with each other), all parameters were found to be statistically not significant (p>0.05). On the other hand, no meaningful difference was found in terms of the serum albumin level. In our opinion, alterations in the level of trace elements and antioxidant proteins, important for many metabolic processes, in laryngeal cancer may not be a reason for but is, in fact, a consequence of the disease itself.

Epidermal growth factor receptors (EGFR) and transferrin receptors (TFR) are known to be involved in cell proliferation and to be expressed in normal human epidermis. To date little is known about EGFR and TRF expression in human skin during embryonic and fetal development. In the present work, we studied skin specimens from 30 aborted embryos and fetuses ranging from 7 to 31 weeks estimated gestational age. Monoclonal antibodies to EGFR and TFR were applied on frozen skin sections using an amplification biotin-streptavidin-fluorescein technique. TFR was faintly expressed on epidermal basal cells throughout embryonic and fetal development, as it is in adult epidermis. Up to week 12, EGFR was uniformly expressed on cells of the basal, intermediate and periderm cell layers. From the midfetal period onwards, the suprabasal cell layers showed a decreased staining compared with the basal layer. During the third trimester the cornified cell layer was completely negative. The hair germ and heir peg cells were positive. Later, the outer root sheath and hair bulb remained labelled, with less staining of the hair cone. The sebaceous and eccrine sweat glands were also labelled. These results suggest that in embryonic and fetal epidermis, TFR expression is not correlated with cellular proliferation, whereas EGFR appear to be associated with proliferating and undifferentiated cells.

To expand the function of DNA machines, we constructed a non-DNA-fuel machine based on the G-quadruplex and i-motif structures within the telomere DNA sequence. Depending on the binding or non-binding of the specified form, the DNA machine is able to bind or release the telomere-binding protein TRF 1, and to release small quadruplex-binding molecules to impede progress of the polymerase. This DNA machine, driven by pH change, does not accumulate duplex DNA waste products to poison the system. These new functions undertaken by structured nucleic acids open many opportunities to create and expand the further functions and use of DNA and RNA.

Recent studies from this laboratory have suggested a similarity, if not identity, of thyrotropin releasing factor (TRF) deamidase and post-proline cleaving enzyme. Bovine brain TRF deamidase was purified to homogeneity and used to elicit antibodies to the enzyme. These antibodies were used to demonstrate identical immunological reactivity between rat brain TRF deamidase and rat kidney post-proline cleaving enzyme. In addition, both proteins exhibit a molecular weight of 75,000, and have identical Km, values for the synthetic substrate pGlu-(N-benzyl-L-His)-Pro-beta-naphthylamide and identical K1 values for TRF and luteinizing hormone releasing factor as inhibitors. Finally, and enzymes exhibit the same sensitivities to inhibition by mercury, iodoacetamide, N-ethylmaleimide, and 5, 5'-dithiobis-(2-nitrobenzoic acid). These results strongly suggest that brain TRF deamidase and kidney post-proline cleaving enzyme are identical.

The aim of this study was to investigate which homogeneous groups, according to teacher reports of attention-deficit/hyperactivity (ADH) Problems on the Teacher's Report Form (TRF), can be identified in a referred sample (n = 4,422; age = 6-18 years; mean age = 9.9 years; 66% boys, 34% girls). Latent class analysis (LCA) was conducted on ADH Problems. In addition, co-morbidity levels in the different ADH Problems groups were compared. LCA yielded three different groups of children and adolescents with both Inattention and hyperactivity-impulsivity, and one group with high scores on Inattention but low scores on hyperactivity-impulsivity. A group of patients with predominantly hyperactivity and impulsivity was not found. Individuals in groups with higher levels of ADH Problems had significantly higher levels of oppositional defiant (OD) and conduct problems, and, although to a lesser extent, significantly higher levels of affective and anxiety problems than individuals in groups with lower levels of ADH Problems. It may not be useful to discern the hyperactive-impulsive type of ADHD.

Maternal smoking during pregnancy (SDP) is a significant public health concern with adverse consequences to the health and well-being of the developing child, including behavioral outcomes such as Attention-Deficit Hyperactivity Disorder (ADHD). There is substantial interest in understanding the nature of this reported association, particularly in light of more recent genetically informed studies that suggest that the SDP-ADHD link is less clear than once thought. In a sample of families (N = 173) specifically selected for sibling pairs discordant for prenatal smoking exposure, we use a sibling-comparison approach that controls for shared genetic and familial influences to assess the effects of SDP on ADHD symptom dimensions. ADHD was measured by both parent and teacher report on the Conners report forms and the Child Behavior Checklist/Teacher Report Form (CBCL/TRF). Results for the CBCL/TRF Total ADHD score are consistent with prior genetically informed approaches and suggest that previously reported associations between SDP and ADHD are largely due to familial confounding rather than causal teratogenic effects. However, results from the Conners parent report suggest a potentially causal effect of SDP on hyperactive/impulsive and, to a lesser extent, total ADHD symptoms; SDP results in increased parent-reported hyperactive/impulsive and total ADHD symptoms even after accounting for genetic and familial confounding factors. This suggests that the Conners assessment (parent-report) may provide a sensitive measure for use in studies examining child specific SDP effects on continuous and dimensional aspects of ADHD. © 2016 Wiley Periodicals, Inc.

OBJECTIVE

The aim of this study was to determine the eff ectiveness and safety of aripiprazole in children and adolescents with both attention deficit/hyperactivity disorder (ADHD) and conduct disorder (CD).

METHODS

20 children and adolescents, ranging in age from 6–16 years, participated in a singlecenter, open-label study (19 to completion). We began treating patients with 2.5 mg of aripiprazole in an open-label fashion for 8 weeks. Outcome measures included the Turgay DSM-IVbased child and adolescent behavior disorders screening and rating scale (T-DSM-IV), the clinical global impressions-severity and improvement scales (CGI-S and CGI-I), the child behavior checklist (CBCL), the teachers report form (TRF) and the extrapyramidal symptom rating scale (ESRS), along with laboratory assessments.

RESULTS

The mean daily dosage of aripiprazole at the end of 8 weeks was 8.55 mg (SD = 1.73), with a maximum dosage of 10 mg. Based on the global improvement subscale of the CGI, we classified 12 of 19 patients (63.1 %) as responders (very much or much improved). We observed significant improvements after aripiprazole treatment with regard to inattention, hyperactivity/impulsivity, ODD, and CD subscales of the T-DSMIV (parent, teacher and clinician forms). We also observed significant improvements on many of the CBCL and TRF subscales (e. g., attention problems as well as delinquent and aggressive behavior). The participants tolerated aripiprazole, and no patient was excluded from the study because of adverse drug events.

CONCLUSIONS

Aripiprazole is an eff ective and well-tolerated treatment for ADHD and CD symptoms; however, additional studies (specifically, placebo-controlled and double-blind studies) are needed to better defi ne the clinical use of aripiprazole in children and adolescents with ADHD-CD.

We have investigated whether the intermediate cells that arise in primed mice after boosting require further cycles of division in culture before maturation into IgG antibody secreting cells. Killing of dividing cells between days 1--4 in culture, by exposure to BUdR-uv irradiation ablated the high IgG response observed on day 5 in control cultures. After T cell removal and replacement by a soluble factor (TRF) similar results were obtained. Thus B cell division over an extended period occurs prior to the appearance of IgG secreting cells. Furthermore, autoradiography of plaques from cultures briefly exposed to [3H]thymidine before harvest showed that some antibody secreting cells were synthesizing DNA at the time of assay.

Free plasma DNA was extracted and terminal restriction fragment (TRF) sequences were amplified by polymerase chain reaction in 32 patients with stage 3 and stage 4 ovarian cancer and 45 healthy controls. Three peaks of TRF were identified and the size of the largest peak (peak 1) correlated with cancer telomere length in cancer patients. Average size of peak 1 in cancer patients was significantly shorter than in controls. When plasma TRF peak 1 was tested pre- and post-treatment, the average pre-treatment size was 8.7 +/- 0.5 Kb, lengthening significantly post-treatment. In long-term survivors of ovarian cancer (10-year disease-free survival), plasma TRF length was the same as in normal controls. The results suggest that the plasma TRF in ovarian cancer patients is of tumour origin. Its determination may reflect tumour cell viability in the host.

It is often difficult to distinguish histologically between an adrenal cortical cancer and a benign adenoma, or to predict the prognosis of patients with adrenal cortical cancers. In this investigation, we examined whether apoptosis-regulating genes, bcl-xL and fas, and a telomere-related gene, telomeric-repeat binding factor-1 (TRF-1), differ between adrenal cortical cancers and benign adrenal tumors. Tissues from 4 adrenal cortical cancers were compared with 7 normal adrenal tissues, 17 cortical adenomas, 4 cortical hyperplasias, and 20 pheochromocytomas for expressions of bcl-xL and fas by RT-PCR, and for expressions of TRF-1 by real-time quantitative RT-PCR. All benign adrenal tissues expressed both the antiapoptosis gene, bcl-xL, and proapoptosis gene, fas, but the adrenal cortical cancers expressed only bcl-xL and not fas. TRF-1 increased by more than 30-fold in the adrenal cortical cancers, compared with benign adrenal tissues, and inversely correlated with the prognosis of patients with the adrenal cortical cancers. This lack of expression of fas in adrenal cortical cancer may help to distinguish it from benign adrenal tumors. The level of TRF-1 expression may be helpful prognostically for patients with adrenal cortical cancers.

OBJECTIVE

In order to explore the role of alterations of telomerase activity and terminal restriction fragment (TRF) length in the development and progression of gastric cancer.

METHODS

Telomerase activity was detected in 176 specimens of gastric mucosa obtained through an operation or endoscopical biopsy by using the telomeric repeat amplification protocol (TRAP) assay. Meanwhile, the mean length of TRF was measured with the use of a Southern blot in part of those samples.

RESULTS

Telomerase activity was detected in 14 of 57 (24.6%) chronic atrophy gastritis patients, six of 18 (33.3%) intestinal metaplasia patients, three of eight (37.5%) dysplasia patients and 60 of 65 (92.3%) gastric cancer patients, respectively. Normal gastric mucosa revealed no telomerase activity. No association was found between telomerase activity and any clinicopathological parameters. The mean TRF length was decreased gradually with age in normal mucosa and in gastric cancer tissue. Regression analysis demonstrated that the reduction rate in these tissues was 41 +/- 12 base pairs/year. Among 35 gastric cancers, TRF length was shown to be shorter in 20 cases (57.1%), similar in 12 cases (34.3%) and elongated in three cases (7.6%), compared to the corresponding adjacent tissues. The mean TRF length tended to decrease as the mucosa underwent chronic atrophy gastritis, intestinal metaplasia, dysplasia and into gastric cancer. The mean TRF length in gastric cancer was not statistically correlated with clinicopathological parameters and telomerase activity.

CONCLUSIONS

Our results suggest that telomerase is expressed during the early stage of gastric carcinogenesis, and that the clinical significance of TRF length appears to be limited in gastric cancer.

We examined the effect of various culture conditions on the polarized secretion of androgen binding protein (ABP) and transferrin (Trf) by Sertoli cells (Sc) in vitro. Sc from 18-day-old rats were cultured as confluent monolayers on permeable membranes in two-compartment chambers for up to 11 days. Coating of the membranes with extracellular matrix (ECM) components: collagen IV + laminin (CL) or reconstituted basement membrane (RBM) enhanced ABP and Trf secretion (200% and 150%, respectively), with RBM being more effective than CL in stimulating ABP but not Trf secretion. Neither CL nor RBM significantly influenced the relative amounts of ABP and Trf secreted into the outer (OC) and inner (IC) compartments of the culture chamber (OC/IC ratio). All of these effects were not significantly influenced by the presence of testosterone and serum. Co-culture of Sc with peritubular myoid cells (Pc) significantly increased the secretion of both ABP and Trf, although the magnitude of stimulation and the time-response patterns were different for each protein. Co-culture with Pc also dramatically increased the OC/IC ratios for ABP and Trf. Testosterone (10(-6) M) enhanced the Pc effects. In cultures of Sc alone, presence of 2% fetal bovine serum increased the OC/IC ratios, whereas testosterone had no effect. Based on these results, we suggest a possible role of Pc in the regulation of Sc polarized secretions.

This study examined the association of low birth weight (LBW) and developmental milestones with behavioral and emotional problems in a general population sample of 3344 Chinese children and adolescents aged 6-16 years in 1997. Parents completed a self-administrated questionnaire including information about birth weight and developmental milestones (i.e. lifting the head up, tooth eruption, speech, walking and bedwetting cessation), and the Child Behavioral Checklist (CBCL). Teachers completed the Teacher's Report Form (TRF) to assess classroom behavior problems. Results indicated that LBW and delayed developmental milestones were significantly associated with an increased risk for almost all parent- and teacher-reported behavioral problems after controlling for the potential effects of child's gender, age and birth order, parental ages at birth, education, occupation, complications at birth and number of children in the family. LBW was significantly associated with delay in achieving all developmental milestones including lifting of the head, tooth eruption, sitting without support, walking without help, speech as saying words with meaning, and bedwetting cessation. It is concluded that LBW and delayed early childhood development may predict the occurrence of a wide range of behavioral and emotional problems in later childhood and adolescence.

The analytical performances of a novel DNA-ligand system using the time-resolved fluorescence (TRF) response of ochratoxin A (OTA)-terbium-DNA aptamer interaction were tested for the quantitative determination of OTA in wheat. Wheat was extracted with acetonitrile/water (60:40, v/v) followed by clean-up through affinity columns containing a DNA-aptamer-based oligosorbent. Then, OTA was detected by TRF spectroscopy after reaction with a terbium fluorescent solution containing the DNA-aptamer probe. The entire procedure was performed in less than 30 min, including sample preparation, and allowed analysis of several samples simultaneously with a 96-well microplate reader. The average recovery from samples spiked with 2.5-25 μg kg(-1) OTA was 77%, with a relative standard deviation lower than 6% and a quantification limit of 0.5 μg kg(-1). Comparative analyses of 29 naturally contaminated (up to 14 μg kg(-1)) wheat samples using the aptamer-affinity column/TRF method or the immunoaffinity column/high-performance liquid chromatography method showed good correlation (r = 0.985) in the range tested. The trueness of the aptamer-based method was additionally assessed by analysis of two quality control wheat materials for OTA. The DNA-ligand system is innovative, simple and rapid, and can be used to screen large quantities of samples for OTA contamination at levels below the EU regulatory limit with analytical performances satisfying EU criteria for method acceptability.

OBJECTIVE

To investigate the relationship between telomere length in peripheral blood white cells and cardiovascular function in a healthy, aging Han Chinese population.

METHODS

In 2012, peripheral blood leukocytes were obtained from 139 healthy individuals in Beijing, China, and telomere restriction fragment (TRF) length was assayed using a digoxigenin-labeled hybridization probe in Southern blot assays. Indicators of cardiovascular function were also evaluated, including electrocardiograms (ECG), (RR, P, PR, QRS, ST and T intervals); blood pressure (BP), (SBP, DBP, PP, PPI); cardiovascular ultrasound (left ventricular ejection fraction, LVEF); mitral early and late diastolic peak flow velocity (MVE and MVA); and lipid indices (TC, TG, HDL, LDL, LCI). The relationships of these cardiovascular indictors to telomere length were evaluated.

RESULTS

No correlations were found between telomere length and ECG, BP or lipid indices even after adjustment for age. Correlations were found between TFR length and some cardiovascular ultrasound indictors (D, MVEA, MVEDT, MVES, MVEL, MVEI, IMT), but these were not seen after adjusting for age.

CONCLUSIONS

We did not find that leukocyte TFR length was associated with cardiovascular ultrasound indictors, ECG, BP, or lipid indices in this population of healthy Han Chinese individuals. Telomere length may serve as a genetic factor in biological aging.