OBJECTIVE

The objective of this study was to determine the normative data and psychometric properties of the parent and teacher rating form of the child behavior checklist (CBCL) in an Iranian community sample.

METHODS

A sample of 6-12 year old students was randomly selected from ten elementary schools in Tehran, Iran. The parent's and teacher's versions of CBCL were accomplished. Clinical interview and the kiddie schedule for affective disorders and schizophrenia - present and lifetime version, Persian version (K-SADS-PL-PV) were used to evaluate the validity and the cut-off point of CBCL and the teacher rating form (TRF).

RESULTS

Among 600 recruited students with mean age of 9.11 years (SD=1.45), 54.16% were girls (n=325). Girls had significantly lower scores in Attention Problems, Delinquent Behavior, Aggressive Behavior, Externalizing and Total Problems than boys (P<0.01). The relation was significant between the CBCL Internalizing and students' ages (β=0.124, P=0.002). The Internal consistency, the correlation among the CBCL and TRF scales, and the inter-rater correlations for CBCL/TRF scales were good to high for most indices and subscales. Based on the receiver operating characteristics (ROC) analysis the best convergences were between the CBCL Attention Problems subscale and attention deficit hyperactivity disorder (ADHD) diagnosis, the CBCL Total Problems and any disorders, the CBCL Externalizing and ADHD+ODD diagnosis. The sensitivities and specificities of the CBCL subscales were higher than the TRF except for Externalizing/ADHD+ oppositional defiant disorder (ODD) which was reverse.

CONCLUSIONS

These results support the multicultural CBCL/TRF findings. CBCL is a useful instrument to consider ADHD and any disorders in community samples.

Restriction in meal timing has emerged as a promising dietary approach for the management of obesity and dysmetabolic diseases. The present systematic review and meta-analysis summarized the most recent evidence on the effect of time-restricted feeding (TRF) on weight-loss and cardiometabolic variables in comparison with unrestricted-time regimens. Studies involving TRF regimen were systematically searched up to January 2019. Effect size was expressed as weighted mean difference (WMD) and 95% confidence intervals (CI). A total of 11 studies, 5 randomized controlled trials and 6 observational, were included. All selected studies had a control group without time restriction; hours of fasting ranged from 12-h until 20-h and study duration from 4 to 8-weeks. Most studies involved the Ramadan fasting. TRF determined a greater weight-loss than control regimens (11 studies, n = 485 subjects) (WMD: -1.07 kg, 95%CI: -1.74 to -0.40; p = 0.002; I² = 56.2%), unrelated to study design. The subgroup analysis showed an inverse association between TRF and fat free mass in observational studies (WMD: -1.33 kg, 95%CI: -2.55 to -0.11; p = 0.03; I² = 0%). An overall significant reduction in fasting glucose concentrations was observed with TRF regimens (7 studies, n = 363 subjects) (WMD: -1.71 mg/dL, 95%CI: -3.20 to -0.21; p = 0.03; I² = 0%), above all in trials (WMD:-2.45 mg/dL, 95%CI: -4.72 to -0.17; p = 0.03; I² = 0%). No between-group differences in the other variables were found. TRF regimens achieved a superior effect in promoting weight-loss and reducing fasting glucose compared to approaches with unrestricted time in meal consumption. However, long-term and well-designed trials are needed to draw definitive conclusions.

High-throughput sequencing studies of small RNAs reveal a complex milieu of noncoding RNAs in biological samples. Early data analysis was often limited to microRNAs due to their regulatory nature and potential as biomarkers; however, many more classes of noncoding RNAs are now being recognized. A class of fragments initially excluded from analysis were those derived from transfer RNAs (tRNAs) because they were thought to be degradation products. More recently, critical cellular function has been attributed to tRNA fragments (tRFs), and their conservation across all domains of life has propelled them into an emerging area of scientific study. The biogenesis of tRFs is currently being elucidated, and initial studies show that a diverse array of tRFs are generated from all parts of a tRNA molecule. The goal of this review was to present what is currently known about tRFs and their potential as biomarkers for the earlier detection of disease.

The main 'acute-phase proteins' were determined in serum of 20 patients with presenile Alzheimer's disease (AD) and compared with values in 18 age-matched healthy control subjects. Following parameters were evaluated: alpha 1-antitrypsin (alpha AT), haptoglobin (HPT), transferrin (TRF), acidic alpha 1-glycoprotein (A alpha G), ceruloplasmin (CER), alpha 2-macroglobulin (alpha MG), C-reactive protein (CRP), albumin (ALB), together with the immunoglobulins IgG, IgM and IgA, and some of the most significant factors of the classic (C3, C4) and alternative (properdin factor B) pathways of complement activation. The results showed a statistically significant increase in the levels of alpha AT (p less than 0.001), CER (p less than 0.001) and of all the complement factors studied (p less than 0.005). The levels of other acute-phase protein (HPT, TRF, A alpha G, alpha MG, CRP, ALB) and immunoglobulins (IgG, IgM, IgA) were similar in AD patients and normal controls. These results give rise the possibility that these elements indicate an altered immunoregulation compatible with chronic cell damage and/or chronic inflammation conditions. Moreover, the increased level of alpha AT can be related to the low production of interleukin-1 (IL-1) reported in AD, which supports the hypothesis of a relative derangement of the macrophage function in presenile AD patients.

BACKGROUND

The dexamethasone suppression test (DST) is the main hormonal disturbance in psychotic depression compared to non-psychotic depression. However, although there have been many studies of individual hormonal axes in depression, few multi-axial studies have been reported. This study aims to examine hormonal differences between these groups of patients through three functional hormonal tests: DST, thyroid stimulating hormone response to thyroid releasing hormone (TSH-TRF) and growth hormone response to growth hormone releasing factor (GH-GRF).

METHODS

Forty inpatients meeting DSM-III-R criteria for major depressive episode with melancholia (21 non-psychotic and 19 psychotic) were studied. Dexamethasone suppression test, TSH-TRF and GH-GRF tests were undertaken for all patients.

RESULTS

In the whole melancholic sample, 80.0% showed disturbances in at least one hormonal axis, 40.0% in two axes and 5.0% in all three axes. Basal and post-dexamethasone cortisol levels were significantly higher in psychotic than in non-psychotic patients. An association between post-dexamethasone cortisol and blunted GH-GRF response was demonstrated in those with psychotic depression. In the whole sample, GH blunting was found in 62.5% of patients, DST non-suppression in 37.5% and TSH blunting in 25.0% (no differences were found between psychotic and non-psychotic patients).

CONCLUSIONS

Sample was restricted to melancholia and unknown factors may influence hormonal responses to stress.

CONCLUSIONS

Hormonal disturbances in depression are more evident when studying several axes, being the HPA and the GH axes the most prominents. Psychotic depression showed more HPA disturbance than non-psychotic depression. Influence of the HPA on the GH axis is discussed.

Eighty child psychiatric inpatients with behavioral and emotional disorders were evaluated from multiple perspectives on admission and at 5-month and 3-year follow-ups. A majority of the patients showed a significant improvement in functioning during the 3-year follow-up. About half of the patients were functioning within clinical range at 3-year follow-up on parental (CBCL) and/or teacher (TRF) ratings. A less favorable outcome was predicted by disruptive behavioral disorder, severity of initial dysfunction, high antisocial and hyperkinetic symptoms, adoptive household and postdischarge institutional placement. Pure anxiety or affective disorder was associated with favorable outcome. Age, sex, place of treatment, and length of hospital treatment were not related to outcome variables.

BACKGROUND

Cardiovascular calcification outside of the coronary tree, known as extracoronary calcification (ECC), is highly prevalent, often occurs concurrently in multiple sites, and yet its prognostic value is unclear.

OBJECTIVE

To determine whether multisite ECC is associated with coronary heart disease (CHD) events, CHD mortality, and all-cause mortality.

METHODS

We evaluated 5903 participants from the Multi-Ethnic Study of Atherosclerosis without diabetes who underwent CT imaging for calcification of the aortic valve, aortic root, mitral valve, and thoracic aorta. Participants were followed for 10.3 years. Multivariable adjusted hazard ratios estimated risk of outcomes for increasing numbers of ECC sites (0, 1, 2, 3, and 4), and receiver operator characteristic analysis assessed model discrimination.

RESULTS

Prevalence of any ECC was 45%; median age was 62 years. Compared with those without ECC, those with ECC in 4 sites had increased hazards of 4.5, 7.1 and 2.3 for CHD events, CHD mortality, and all-cause mortality, respectively, independent of traditional risk factors (TRF; all P ≤ .05), and had ≥2-fold increased hazards for outcomes independent of coronary artery calcification (CAC). Each additional site of ECC was positively associated with each outcome in a graded fashion. When added to TRF, ECC significantly increased the area under the receiver operator characteristic curve for all outcomes and modestly increased the area under the curve for mortality beyond TRF + CAC (0.799 to 0.802; P = .03).

CONCLUSIONS

Increasing multisite ECC has a graded association with higher CHD and mortality risk, contributing information beyond TRF. Multisite ECC incidentally identified on imaging can be used to improve individualized risk prediction.

This study aimed to investigate early lexical and grammatical development and their relations in a sample of very immature healthy preterms, in order to assess whether their linguistic development was typical, at risk or atypical. The effects of biological factors and parental level of education on preterms' linguistic development were also investigated. Seventy-three Italian preterms and 22 Italian fullterms (control sample) were assessed at 2;6 with an Italian test of repetition of sentences (TRF). Their mothers completed the Italian version of the MacArthur questionnaire (PVB). Our results showed that most of the preterm sample displayed a typical development, compared with the fullterms, with lexicon and grammar within the normal range and a normal relation between these competencies. However, preterms characterized by an extremely low birthweight (ELBW), a very low gestational age and male gender were at risk, with slight delays in the lexicon and grammar, but still retaining the normal relation between the two.

Skeletal muscle satellite cells are heavily involved in the regeneration of skeletal muscle in response to the aging-related deterioration of the skeletal muscle mass, strength, and regenerative capacity, termed as sarcopenia. This study focused on the effect of tocotrienol rich fraction (TRF) on regenerative capacity of myoblasts in stress-induced premature senescence (SIPS). The myoblasts was grouped as young control, SIPS-induced, TRF control, TRF pretreatment, and TRF posttreatment. Optimum dose of TRF, morphological observation, activity of senescence-associated β-galactosidase (SA-β-galactosidase), and cell proliferation were determined. 50 μg/mL TRF treatment exhibited the highest cell proliferation capacity. SIPS-induced myoblasts exhibit large flattened cells and prominent intermediate filaments (senescent-like morphology). The activity of SA-β-galactosidase was significantly increased, but the proliferation capacity was significantly reduced as compared to young control. The activity of SA-β-galactosidase was significantly reduced and cell proliferation was significantly increased in the posttreatment group whereas there was no significant difference in SA-β-galactosidase activity and proliferation capacity of pretreatment group as compared to SIPS-induced myoblasts. Based on the data, we hypothesized that TRF may reverse the myoblasts aging through replenishing the regenerative capacity of the cells. However, further investigation on the mechanism of TRF in reversing the myoblast aging is needed.

OBJECTIVE

Osteoporosis is one of the common orthopedic diseases featured in low bone mineral density. Exosomes have been proven to be potential markers for many diseases and health problems. The roles of messenger RNAs and microRNAs in osteoporosis have been comprehensively studied; however, little research has focused on the function of plasma exosomal transfer RNA-derived fragments (tRFs) in osteoporosis.

METHODS

We collected plasma samples from 40 healthy controls and 40 osteoporosis patients, and all exosomes were isolated with combined centrifugation and were characterized by electron microscopy. Small RNA sequence (Yingbio) was performed to detect the plasma exosomal tRFs and tRF markers were validated by real-time quantitative polymerase chain reaction (qPCR). Three exosome diagnostic tRFs were confirmed by receiver operating characteristic analyses.

RESULTS

In this study, 11 upregulated tRFs and 18 downregulated tRFs were identified in osteoporosis compared with normal controls. Higher expression levels of plasma exosomal tRF-25-R9ODMJ6B26 (tRF-25), tRF-38-QB1MK8YUBS68BFD2 (tRF-38), tRF-18-BS68BFD2 (tRF-18) in osteoporosis were confirmed by qPCR. Plasma exosomal tRF-25, tRF-38 and tRF-18 showed better accuracy for osteoporosis diagnosis.

CONCLUSIONS

Our results suggest that plasma exosomal tRF-25, tRF-38 and tRF-18 might be diagnostic biomarkers for osteoporosis detection.

We aimed at evaluating the relationship between lean mass and fat mass with age, menopausal age (MA) and years since menopause (YSM) and their effects on bone mineral density (BMD) at segmental regions in postmenopausal elderly women with and without osteoporosis. After using a dual-energy X-ray absorptiometry (DXA) methodology to measure body composition and BMD at posteroanterior spine and hip in 244 postmenopausal elderly non-osteoporotic (Non-OP) women (65.5 ± 4.3 years) and 298 postmenopausal elderly osteoporotic (OP) women (67.1 ± 4.4 years), we found that in postmenopausal elderly Non-OP women, there was no correlation between lean mass with age, MA, and YSM, as well as no correlation between fat mass with age (all, p>> 0.05); leg fat (LF) mass (r = 0.187; p<0.01), whole body fat (WF) mass (r = 0.151; p < 0.05), and trunk fat (TRF) mass (r = 0.141; p < 0.05) were positively correlated with MA; LF (r = -0.131; p < 0.05) and WF (r = -0.127; p < 0.05) were negatively associated with YSM; WF and whole body lean (WL) mass were the most important body composition components influencing BMD at the third lumbar spine (L3), total first to fourth lumbar spine (L1-4) and hip, respectively; TRF was the most significant determinant of BMD at both L2 and L4. In postmenopausal elderly OP women, there was no relationship between body composition with MA (p>> 0.05); Trunk lean (TRL) mass (r = -0.183; p < 0.05), leg lean (LL) mass (r = -0.136; p < 0.01), and WL mass (r = -0.162; p < 0.01) were negatively correlated with age; TRL mass (r = -0.132; p < 0.05), LL mass (r = -0.152; p < 0.01), WL mass (r = -0.170; p < 0.01) were also negative with YSM; WF was the most important factor influencing BMD at lumbar spine and hip. These data suggest in postmenopausal elderly Non-OP women, fat mass (TRF, LF, and WF) was more related with MA; WF and WL mass were the most important body composition components influencing BMD at L1-4 and hip, respectively; in postmenopausal elderly OP women, body composition was not correlated with MA; lean mass (TRL, LL, and WL) was more age-related negatively; WF mass was the most significant factor affecting BMD at lumbar spine and hip.

Chronic lymphocytic leukemia (CLL) is the most common human leukemia, and dysregulation of tRNA-derived short noncoding RNA (tsRNA) (tRF-1) expression is an accompanying event in the development of this disease. tsRNAs are fragments originating from the 3' end of tRNA precursors and do not contain mature tRNA sequences. In contrast to tsRNAs, mature tRFs (tRF-3s, tRF-5s, and internal tRFs) are produced from mature tRNA sequences and are redundant fragments. We investigated tsRNA expression in CLL and determined tsRNA signatures in indolent CLL and aggressive CLL vs. normal B cells. We noticed that both ts-43 and ts-44 are derived from distinct genes of pre-tRNA^His, and are down-regulated in CLL 3- to 5-fold vs. normal B cells. Thus, we investigated expression levels of tRF-5 fragments from tRNA^His in CLL samples and healthy controls, and determined that such fragments are down-regulated by 5-fold in CLLs vs. normal controls. Given these results, we investigated the expression of all mature tRFs in CLLs vs. normal controls. We found a drastic dysregulation of the expression of mature tRFs in CLL. In aggressive CLL, for the top 15 up-regulated fragments, linear fold change varied from 2,053- to 622-fold. For the top 15 down-regulated fragments in CLL, linear fold change varied from 314- to 52-fold. In addition, 964 mature tRFs were up-regulated at least 2-fold in CLL, while 701 fragments were down-regulated at least 2-fold. Similar results were obtained for indolent CLL. Our results suggest that mature tRFs may have oncogenic and/or tumor suppressor function in CLL.

BACKGROUND

The Strengths and Difficulties Questionnaire (SDQ) is a valuable screening tool for identifying psychosocial problems. Its performance in a multi-ethnic society, common to many paediatric health care workers, has not been investigated. Because it is important that screening instruments are valid and reliable for all ethnic groups within one society, we examined differences in the SDQ's psychometric properties in a multi-ethnic society.

METHODS

The SDQ parent (n = 8114) and teacher form (n = 9355) were completed as part of a preventive health check for children aged 5-6 years of Dutch and non-Dutch ethnic backgrounds. The Child Behaviour Checklist (CBCL)/Teacher Report Form (TRF) was administered to a subsample.

RESULTS

Factor analysis of the parent-rated SDQ showed different rating patterns for two of the five subscales for non-Dutch children as compared with Dutch children. Cronbach's alpha for the total difficulties score varied by ethnic group (0.73-0.78 parent-rated SDQ, 0.80-0.83 teacher-rated SDQ), and coefficients were generally smaller for non-Dutch than for Dutch children (P < 0.05). Alpha coefficients for subscales varied between 0.31-0.85 for ethnic groups. Inter-rater correlations between parents and teachers for the total difficulties score varied between 0.20-0.41 between ethnic groups and were larger for Dutch than for non-Dutch children (P < 0.05). Concurrent validity was acceptable for most scales and most ethnic groups.

CONCLUSIONS

The total difficulties score of the parent- and teacher-rated SDQ is valid and reliable for different ethnic groups within Dutch society. However, there are differences in reliability and validity of the subscales, which makes interpretation of the subscales difficult for certain ethnic groups.

Lung cancer is the most prevalent and deadliest cancer worldwide. A significant part of lung cancer studies is dedicated to the expression alterations of non-coding RNAs. The non-coding RNAs are transcripts that cannot be translated into proteins. While the study of microRNAs and siRNAs in lung cancer received a lot of attention over the last decade, highly efficient therapeutic option or the diagnostic methods based on non-coding RNAs are still lacking. Because of this, it is of utmost importance to direct future research on lung cancer towards analyzing other RNA types for which the currently available data indicates that are essential at modulating lung tumorigenesis. Through our review of studies on this subject, we identify the following non-coding RNAs as tumor suppressors: ts-46, ts-47, ts-101, ts-53, ts-3676, ts-4521 (tRNA fragments), SNORD116-26, HBII-420, SNORD15A, SNORA42 (snoRNAs), piRNA-like-163, piR-35127, the piR-46545 (piRNAs), CHIAP2, LOC100420907, RPL13AP17 (pseudogenes), and uc.454 (T-UCR). We also found non-coding RNAs with tumor-promoting function: tRF-Leu-CAG, tRNA-Leu, tRNA-Val (tRNA fragments), circ-RAD23B, circRNA 100146, circPVT1, circFGFR3, circ_0004015, circPUM1, circFLI1, circABCB10, circHIPK3 (circRNAs), SNORA42, SNORA3, SNORD46, SNORA21, SNORD28, SNORA47, SNORD66, SNORA68, SNORA78 (snoRNAs), piR-65, piR-34871, piR-52200, piR651 (piRNAs), hY4 5' fragments (YRNAs), FAM83A-AS1, WRAP53, NKX2-1-AS1 (NATs), DUXAP8, SFTA1P (pseudogene transcripts), uc.338, uc.339 (T-UCRs), and hTERC.

The present study examines the deleterious effect of biologically synthesized silver nanoparticles in adult zebrafish. Silver nanoparticles (AgNPs) used in the study were synthesized by treating AgNO3 with aqueous leaves extract of Malva crispa Linn., a medicinal herb as source of reductants. LC50 concentration of AgNPs at 96 h was observed as 142.2 μg/l. In order to explore the underlying toxicity mechanisms of AgNPs, half of the LC50 concentration (71.1 μg/l) was exposed to adult zebrafish for 14 days. Cytological changes and intrahepatic localization of AgNPs were observed in gills and liver tissues respectively, and the results concluded a possible sign for oxidative stress. In addition to oxidative stress the genotoxic effect was observed in peripheral blood cells like presence of micronuclei, nuclear abnormalities and also loss in cell contact with irregular shape was observed in liver parenchyma cells. Hence to confirm the oxidative stress and genotoxic effects the mRNA expression of stress related (MTF-1, HSP70) and immune response related (TLR4, NFKB, IL1B, CEBP, TRF, TLR22) genes were analyzed in liver tissues and the results clearly concluded that the plant extract mediated synthesis of AgNPs leads to oxidative stress and immunotoxicity in adult zebrafish.

A large proportion of the population in Uganda still relies on the use of plant extracts for treatment of various ailments. This study tested the claimed efficacy of some plants in the treatment of measles. In vitro antiviral assays were performed on extracts of two medicinal plants (Warburgia ugandensis and Zanthoxylum chalybeum) using measles virus (Edmonston and Swartz strains) as the test organisms. The assays performed were the neutralisation tests and the plaque reduction assays. Of the two plants Z. chalybeum had demonstrable in vitro antiviral activity in the seed extracts (titer reduction factor [TRF]: 100, for the ethanolic extract). The in vitro antiviral activity of the seed extracts was demonstrated to be due to compound 27-135D (TRF=1000), which was characterized by (1)H-NMR spectroscopy as the alkaloid skimmianine. Skimmianine had minimal toxicity to VERO cell lines. The petroleum ether extracts and the ethanolic extracts of Warburgia ugandensis had no inhibitory effect on cytopathic effect (CPE) formation, especially at the maximal non-toxic dose (MNTD). The extracts of W. ugandensis were highly toxic to VERO cell lines. The TRF values for the stem bark extracts of W. ugandensis were: water extract, 10; ethanolic extract, 1; fraction 27-163D, 100., which were regarded to be too low. Seed extracts of Z.chalybeum therefore probably cure measles due to the antiviral effect of skimmianine. It is not clear how extracts of W. ugandensis produce a beneficial response in measles disease, if at all.

This study compared the ability of three forms of vitamin E [tocotrienol-rich fraction (TRF), alpha-tocopherol (α-T), and delta-tocotrienol (δ-T3)] to enhance immune response to tetanus toxoid (TT) immunisation in a mouse model. Twenty BALB/c mice were divided into four groups of five mice each. The mice were fed with the different forms of vitamin E (1 mg) or vehicle daily for two weeks before they were given the TT vaccine [4 Lf] intramuscularly (i.m.). Booster vaccinations were given on days 28 and 42. Serum was collected (days 0, 28, and 56) to quantify anti-TT levels. At autopsy, splenocytes harvested were cultured with TT or mitogens. The production of anti-TT antibodies was augmented (P < 0.05) in mice that were fed with δ-T3 or TRF compared to controls. The production of IFN-γ and IL-4 by splenocytes from the vitamin E treated mice was significantly (P < 0.05) higher than that from controls. The IFN-γ production was the highest in animals supplemented with δ-T3 followed by TRF and finally α-T. Production of TNF-α was suppressed in the vitamin E treated group compared to vehicle-supplemented controls. Supplementation with δ-T3 or TRF can enhance immune response to TT immunisation and production of cytokines that promote cell-mediated (TH1) immune response.

We examined the prevalence of emotional and behavioral problems and associated risk and protective factors among children and adolescents ages 6 to 18 years reared in orphanages in Turkey (n = 461, 87.9% of all eligible subjects) compared with a nationally representative community sample of similarly-aged youngsters brought up by their own families (n = 2280). Using the 90th percentile as the cut-off criterion, it was found that the Teacher's Report Form (TRF) Total Problem score was higher for children and adolescents in orphanage care than in the community (23.2%, orphanage v. 11%, community). Multiple regression models explained 73% of the total variance of TRF Total Problems score for children and adolescents in orphanages. Regular contact with parents or relatives, between classroom teachers and orphanage staff, appropriate task involvement, perceived social support and competency were significant protective factors against emotional and behavioral problems. Younger age at first admission, being small for age, and feelings of stigmatization were associated with higher TRF Problem Scores (P<.05). Parental psychiatric disorder was unrelated to emotional and behavioral problems in children reflecting that psychosocial adversity and parenting problems in of themselves lead to institutionalization, irrespective of identifiable parental mental disorder. The findings are interpreted in the light of an urgent need for development of early intervention programs that promote community care of children by preventing separation from families, provision of support services for families in need, and development of counseling programs to prevent abandonment, abuse and neglect. Finding ways for child welfare professionals to collaborate more closely with early intervention programs would also increase the viable opportunities and rights of children and adolescents currently cared for in the system. Finally, alternative cost-effective care models need to be promoted including foster care or adoption systems and family based homes in the community.

GPCRs represent important targets for drug discovery because GPCRs participate in a wide range of cellular signaling pathways that play a role in a variety of pathological conditions. A large number of screening assays have been developed in HTS laboratories for the identification of hits or lead compounds acting on GPCRs. One type of assay that has found relatively widespread application, due to its at least in part generic nature, relies on the use of a radioactive GTP analogue, [(35)S]GTPgammaS. The G-protein alpha subunit is an essential part of the interaction between receptor and G proteins in transmembrane signaling, where the activated receptor catalyzes the release of GDP from Galpha, thereby enabling the subsequent binding of GTP or a GTP analogue. [(35)S]GTPgammaS allows the extent of this interaction to be followed quantitatively by determining the amount of radioactivity associated with cell membranes. However, with the increased desire to move assays to nonradioactive formats, there is a considerable need to develop a nonradioactive GTP binding assay to monitor ligand-induced changes in GPCR activity. The Eu-GTP binding assay described here is based on TRF that exploits the unique fluorescence properties of lanthanide chelates, and provides a powerful alternative to assays using radioisotopes. In this article, we have used the human alpha(2A)-AR as a model GPCR system to evaluate the usefulness of this Eu-GTP binding assay.

Format and instructional revisions were made in the TRF, a previously reported school adjustment measure, to extend its diagnostic, prescriptive, and empirical utility. Factor analyses of school adjustment ratings on the revised CARS with a normative sample of "healthy" primary graders demonstrated that while the revisions maintained the scale's original three-factor structure, they increased specific item factor loadings and accounted scale variance. To extend the screening utility of the proposed measure, normative and parametric comparisons are reported describing adjustment ratings for sex, age, and city/country subgroups. Comparisons of children who are or are not referred to a secondary prevention program provide evidence of the CARS discriminative validity and screening potential.

Changes in mean telomeric terminal restriction fragment (TRF) length were examined as a marker for cellular replicative history in HIV-1-infected individuals after institution of anti-retroviral therapy (ART). Increases in mean T cell TRF lengths were observed in most patients following therapy; however, the contribution of individual T cell subsets was complex. An elongation of CD8+ T cell TRF was nearly uniformly observed while changes in mean TRF length in CD4+ T cells were heterogeneous as, despite potent suppression of viral replication, CD4 cell telomeres recovered in some patients, yet continued to decline in others. Increases in CD8 cell TRF correlated with decreased memory cells, suggesting a negative selection in the periphery for CD8 cells with extensive replicative history. In contrast, increases in CD4+ T cell TRF length correlated with increases in naive cell subsets, suggesting that the CD4+ T cell TRF increase may reflect a thymic contribution in some patients. These are the first increases in somatic cell telomere length in a population of cells observed in vivo, and the findings are compatible with therapy-induced reconstitution of the lymphoid compartment with cells having a more extensive replicative potential. These findings further distinguish lymphocytes from other somatic cell populations where only decreases in TRF over time have been noted. Thus, institution of ART in persons with moderately advanced HIV-1 disease reveals distinct population dynamics of CD4 and CD8 T cell subsets and also shows that the lymphocyte replicative history is dynamic.

OBJECTIVE

The α-tocopherol and tocotrienol-rich fraction (TRF) are considered effective antioxidants. This study aimed to compare the antioxidative and antifibrotic effects of α-tocopherol and TFR in dibutylin dichloride (DBTC)-induced chronic pancreatitis (CP) rats.

METHODS

Oral administration of α-tocopherol and TFR (both 800 mg/kg per day) started the next day after DBTC (8 mg/kg) infusion into the tail vein for 4 weeks. Histological examination, Sirius red staining, and measurement of the contents of hydroxyproline and malondialdehyde of the pancreas were performed to evaluate pancreatic damage and fibrosis. Immunohistochemical analysis of α-smooth muscle actin and real-time reverse transcription polymerase chain reaction for transforming growth factor-β1 (TGF-β1) and collagen-α1(I) were performed to evaluate the activation of pancreatic stellate cells and the mRNA levels of fibrosis-related genes, respectively.

RESULTS

Both α-tocopherol and TRF reduced oxidative stress, ameliorated inflammation and fibrosis, and down-regulated the mRNA expression of TGF-β1 and collagen-α1(I) in DBTC-induced CP. The TRF was superior to α-tocopherol in alleviating inflammation and fibrosis and down-regulating TGF-β1 mRNA expression.

CONCLUSIONS

Oral administration of α-tocopherol and TRF improves pancreatic inflammation and fibrosis in DBTC-induced CP rats, with TRF being more effective than α-tocopherol. Therefore, TRF may be a novel option for alleviating inflammation and, particularly, the fibrotic process in CP.

A T-cell replacing factor (TRF)/interleukin-5 (IL-5) is a B-cell growth and differentiation factor. In the present study, we examined the role of TRF/IL-5 in the increase in the levels of interleukin-2 (IL-2) receptor expression on activated B-cells. High pressure liquid chromatography (HPLC)-purified TRF/IL-5 (B151-TRF) from TRF-producing T-cell hybridoma, B151K12, as well as recombinant TRF/IL-5 (rec-TRF) were used for the analysis. Maximum anti-2,4-dinitrophenyl (DNP) IgG antibody response of DNP-primed B-cells or polyclonal IgM secretion of B-cell tumor line BCL1 was seen when HPLC-purified B151-TRF was added or when suboptimal doses of B151-TRF were added to the culture in the presence of IL-2. Normal resting B-cells gave maximum anti-SRBC IgM PFC responses when HPLC-purified B151-TRF and IL-2 were present. The purified B151-TRF as well as rec-TRF also induced on B-cells increased expression of IL-2 receptors that react with monoclonal anti-murine IL-2 receptor antibody, PC61, and 125I-labelled IL-2. The numbers of functional high affinity IL-2 receptors on activated B cells increased at least 20-fold by culturing them with purified B151-TRF. Moreover, B151-TRF induced increase in the levels of steady-state mRNA for IL-2 receptor by approximately 8-fold. These results suggest that activated B-cells as well as BCL1-cells may express functional IL-2 receptors or closely related molecules when stimulated with HPLC-purified B151-TRF as well as rec-TRF.