Miracle-dz/UniFAR-FTUs huggingface dataset:
Estradiol and its metabolites 4‐hydroxyestradiol and 2‐hydroxyestradiol induce mutations in human breast epithelial cells

An elevated incidence of breast cancer in women has been associated with prolonged exposure to high levels of estrogens. Our laboratory demonstrated that treatment of the immortalized human breast epithelial cells MCF‐10F with 17β‐estradiol (E2), 4‐hydroxyestradiol (4‐OHE2) or 2‐hydroxyestradiol (2‐OHE2) induces phenotypical changes indicative of neoplastic transformation. MCF‐10F cells treated with E2, 4‐OHE2 or 2‐OHE2 formed colonies in agar methocel and lost their ductulogenic capacity in collagen, expressing phenotypes similar to those induced by the carcinogen benzo[a]pyrene. To investigate whether the transformation phenotypes were associated with genomic changes, cells treated with E2, 4‐OHE2 or 2‐OHE2 at different doses were analyzed using microsatellite markers. Since microsatellite instability (MSI) and loss of heterozygosity (LOH) in chromosomes 13 and 17 have been reported in human breast carcinomas, we tested these parameters in MCF‐10F cells treated with E2, 2‐OHE2, or 4‐OHE2 alone or in combination with the antiestrogen ICI182780. MCF‐10F cells treated with E2 or 4‐OHE2, either alone or in combination with ICI182780, exhibited LOH in the region 13q12.3 with the marker D13S893 located at ∼0.8 cM telomeric to BRCA2. Cells treated with E2 or 4‐OHE2 at doses of 0.007 and 70 nM and 2‐OHE2 only at a higher dose (3.6 μM) showed a complete loss of 1 allele with D13S893. For chromosome 17, differences were found using the marker TP53‐Dint located in exon 4 of p53. Cells treated with E2 or 4‐OHE2 at doses of 0.007 nM and 70 nM and 2‐OHE2 only at a higher dose (3.6 μM) exhibited a 5 bp deletion in p53 exon 4. Our results show that E2 and its catechol estrogen metabolites are mutagenic in human breast epithelial cells. ICI182780 did not prevent these mutations, indicating that the carcinogenic effect of E2 is mainly through its reactive metabolites 4‐OHE2 and 2‐OHE2, with 4‐OHE2 and E2 being mutagenic at lower doses than 2‐OHE2. © 2005 Wiley‐Liss, Inc.
Race/ethnicity, life-course socioeconomic position, and body weight trajectories over 34 years: the Alameda County Study.

OBJECTIVES We investigated whether race differences in weight gain over 34 years were because of socioeconomic position (SEP) and psychosocial and behavioral factors (physical activity, cigarette smoking, alcohol consumption, depression, marital status, number of children). We used a life-course approach to SEP with 4 measures of SEP (childhood SEP, education, occupation, income) and a cumulative measure of SEP. METHODS We used mixed models and data collected from the Alameda County Study to examine the association between race and weight change slopes and baseline weight in men (n=1186) and women (n=1375) aged 17 to 40 years at baseline (in 1965). RESULTS All subjects gained weight over time. African American women weighed 4.96 kg (P < .001) more at baseline and gained 0.10 kg/year (P = .043) more weight than White women. Black men weighed 2.41 kg (P = .006) more at baseline but did not gain more weight than White men. The association of race with weight gain in women was largely because of cumulative SEP score. CONCLUSIONS Interventions to prevent overweight and obesity should begin early in life and target the socioeconomically disadvantaged.
P300 acetyltransferase regulates fatty acid synthase expression, lipid metabolism and prostate cancer growth

De novo fatty acid (FA) synthesis is required for prostate cancer (PCa) survival and progression. As a key enzyme for FA synthesis fatty acid synthase (FASN) is often overexpressed in human prostate cancers and its expression correlates with worse prognosis and poor survival. P300 is an acetyltransferase that acts as a transcription co-activator. Increasing evidence suggests that P300 is a major PCa promoter, although the underlying mechanism remains poorly understood. Here, we demonstrated that P300 binds to and increases histone H3 lysine 27 acetylation (H3K27Ac) in the FASN gene promoter. We provided evidence that P300 transcriptionally upregulates FASN expression and promotes lipid accumulation in human PCa cells in culture and Pten knockout prostate tumors in mice. Pharmacological inhibition of P300 decreased FASN expression and lipid droplet accumulation in PCa cells. Immunohistochemistry analysis revealed that expression of P300 protein positively correlates with FASN protein levels in a cohort of human PCa specimens. We further showed that FASN is a key mediator of P300-induced growth of PCa cells in culture and in mice. Together, our findings demonstrate P300 as a key factor that regulates FASN expression, lipid accumulation and cell growth in PCa. They also suggest that this regulatory pathway can serve as a new therapeutic target for PCa treatment.
Clinical Characteristics of 362 Patients with Familial Migraine with Aura

The objectives of the present study were to describe the clinical characteristics of patients with severe familial non-hemiplegic migraine with aura (NHMA) and to compare these data to those from cases in previous population-based Danish studies using the same methodology. NHMA families were recruited from the Danish patient registry and from Danish neurology practices. A total of 362 NHMA patients were diagnosed according to the 1988 International Headache Society criteria using a validated semistructured physician-conducted interview. Visual aura occurred in almost every NHMA attack. In aura without headache visual aura occurred primarily in isolation. Aura without headache was most common in older, male patients. Several clinical characteristics of familial NHMA differed from migraine with aura in the general population: firstly, the age at onset was lower, secondly, the age at cessation was higher, thirdly, aura symptoms were more severe and finally, the co-occurrence of migraine without aura was higher in familial NHMA. There seems to be a correlation between more severe symptoms and familial aggregation. These results have both clinical and scientific implications.
Myosin VI Is Differentially Regulated by DNA Damage in p53- and Cell Type-dependent Manners*

Myosin VI is an unconventional motor protein and functions in a variety of intracellular processes such as cell migration, vesicular trafficking, and homeostasis of the Golgi complex. Previously, we found that myosin VI is up-regulated in RKO, LS174T, and H1299 cells by DNA damage in a p53-dependent manner and mediates the pro-survival function of p53. Here, we showed that the levels of myosin VI protein were markedly inhibited in MCF7 and LNCaP cells by topoisomerase I-II inhibitors. However, the levels of myosin VI transcript were decreased only by topoisomerase I inhibitors. We also found that the levels of myosin VI protein were markedly inhibited in MCF7 cells by wild-type p53 but not tumor-derived mutant p53. Surprisingly, we found that the level of myosin VI transcript was slightly increased instead of decreased in MCF7 cells by p53, suggesting that a mechanism other than transcriptional repression is involved. Additionally, we found that on the myosin VI promoter, the level of acetylated histone H3 was markedly decreased, whereas that of p53 and acetylated histone H4 was slightly increased in MCF7 cells upon treatment with topoisomerase I-II inhibitors. Finally, we showed that overexpression of myosin VI enhances, whereas knockdown of myosin VI decreases, DNA damage-induced stabilization of p53, and consequently, knockdown of myosin VI de-sensitizes MCF7 cells to DNA damage-induced apoptosis. Taken together, as a mediator of the p53 pro-survival pathway and a marker of malignancy in some tumors, differential regulation of myosin VI in various tumor cells by topoisomerase inhibitors dictates whether knockdown of myosin VI inhibits, rather than enhances, the susceptibility of tumor cells to some therapeutic agents, which might be explored for designing a proper therapeutic strategy.
Modulation of IL-8 and RANTES Release in Human Conjunctival Epithelial Cells: Primary Cells and Cell Line Compared and Contrasted

Purpose. Recent research indicates that epithelial cells of the ocular surface can contribute to the allergic reaction by the release of inflammatory and/or chemotactic mediators. In this study, the role of two inflammatory mediators, previously identified in the tear film of ocular allergy subjects, TNF-&agr; and IFN-&ggr;, were evaluated for their effect on the release of two chemotactic mediators, IL-8 and RANTES, from cultured human conjunctival epithelial cells. Methods. Human conjunctival epithelial cells (primary cells or HC0597 cell line) were grown to confluence and stimulated with various concentrations of TNF-&agr;, IFN-&ggr;, or a combination of both. Supernatants were collected at 6, 24, and 48 hours and stored frozen for subsequent ELISA analyses of RANTES and IL-8. Results. RANTES and IL-8 release from HC0597 cells was stimulated in a dose- and time-dependent manner following treatment with TNF-&agr;. However, only RANTES release was modulated by IFN-&ggr; treatment. Treatment of HC0597 cells with both TNF-&agr; and IFN-&ggr; resulted in a synergistic increase in the release of RANTES. This synergistic effect was confirmed using primary cultures of human conjunctival epithelial cells. Conclusions. Stimulation of conjunctival epithelium with proinflammatory mediators, TNF-&agr; and/or IFN-&ggr;, generated the release of the chemotactic factors IL-8 and RANTES, which could act to prolong inflammation. These two chemokines may prolong inflammation by recruiting eosinophils to the ocular surface. This is the first study to compare chemokine release in a cell line and primary cells; similar chemokine release after mediator stimulation was demonstrated, indicating that the two cell types are phenotypically similar.
Plant Morphological and Biochemical Responses to Field Water Deficits : II. Responses of Leaf Glycerolipid Composition in Cotton.

The effects of water deficits on leaf glycerolipid composition were analyzed in two photoperiodic strains of field grown cotton (Gossypium kirsutum L.) that differ in sensitivity to drought. Leaves from plants grown under dryland conditions exhibited increased dry weight and specific leaf weight. The average midday leaf water potential in the dryland treatment decreased to -1.9 and -2.4 megapascals, respectively, for the T25 and T185 genotypes. Total leaf lipid content of plants exposed to dryland conditions was 5.9 and 7.5% of leaf dry weight for strain T25 and T185, respectively. The difference in leaf lipid content between these genotypes was caused by water deficits and was attributed to loss of both phospholipids and glycolipids in strain T25. There was no apparent loss of phospholipids due to water deficits in the T185 genotype; however, a significant loss of glycolipids was partially compensated by a 2-fold increase in triacylglycerol. No change in triacylglycerol was found between treatments in T25 leaves. Water deficit caused a significant decline in the relative degree of acylunsaturation in phospholipids and glycolipids from both genotypes; however, the double bond index for triacylglycerol increased in both genotypes. It is believed that the observed responses of leaf lipid composition to dryland conditions may be an additional criterion for characterization and selection of new drought-tolerant cotton genotypes.
Protective antigen‐binding domain of anthrax lethal factor mediates translocation of a heterologous protein fused to its amino‐ or carboxy‐terminus

The edema factor (EF) and lethal factor (LF) components of anthrax toxin require anthrax protective antigen (PA) for binding and entry into mammalian cells. After internalization by receptor‐mediated endocytosis, PA facilitates the translocation of EF and LF across the membrane of an acidic intracellular compartment. To characterize the translocation process, we generated chimeric proteins composed of the PA recognition domain of LF (LFN; residues 1–255) fused to either the amino‐terminus or the carboxy‐terminus of the catalytic chain of diphtheria toxin (DTA). The purified fusion proteins retained ADP‐ribosyltransferase activity and reacted with anti‐sera against LF and diphtheria toxin. Both fusion proteins strongly inhibited protein synthesis in CHO‐K1 cells in the presence of PA, but not in its absence, and they showed similar levels of activity. This activity could be inhibited by adding LF or the LFN fragment (which blocked the interaction of the fusion proteins with PA), by adding inhibitors of endo‐some acidification known to block entry of EF and LF into cells, or by introducing mutations that attenuated the ADP‐ribosylation activity of the DTA moiety. The results demonstrate that LFN fused to either the amino‐terminus or the carboxy‐terminus of a heterologous protein retains its ability to complement PA in mediating translocation of the protein to the cytoplasm. Besides its importance in understanding translocation, this finding provides the basis for constructing a translocation vector that mediates entry of a variety of heterologous proteins, which may require a free amino‐ or carboxy‐terminus for biological activity, into the cytoplasm of mammalian cells.
Diet-Induced Obesity and Its Differential Impact on Periodontal Bone Loss

Obesity is associated with abnormal lipid metabolism and impaired bone homeostasis. The aim of our study was to investigate the impact of specific elevated fatty acid (FA) levels on alveolar bone loss in a Porphyromonas gingivalis–induced model of periodontal disease and to analyze underlying cellular mechanisms in bone-resorbing osteoclasts and bone-forming osteoblasts in mice. Four-week-old male C57BL/6 mice were randomly divided in groups and subjected to a palmitic acid (PA)– or oleic acid (OA)–enriched high-fat diet (HFD) (20% of calories from FA) or a normal caloric diet (C group) (10% of calories from FA) for 16 wk. Starting at week 10, mice were infected orally with P. gingivalis (W50) or placebo to induce alveolar bone loss. Animals were sacrificed, and percentage fat, serum inflammation (tumor necrosis factor [TNF]-α), and bone metabolism (osteocalcin [OC], carboxy-terminal collagen crosslinks [CTX], and N-terminal propeptides of type I procollagen [P1NP]) markers were measured. Osteoblasts and osteoclasts were cultured in the presence of elevated PA or OA levels and exposed to P. gingivalis. Animals on FA-enriched diets weighed significantly more compared with animals on a normal caloric diet (P < 0.05). Both obese groups had similar percentages of fat (P = nonsignificant); however, alveolar bone loss was significantly greater in animals that were on the PA-enriched HFD (P < 0.05). TNF-α levels were highest in the PA group (P < 0.001) and increased in all groups in response to P. gingivalis inoculation (P < 0.01), whereas bone remodeling markers OC, CTX, and P1NP were lowest in the PA group (P < 0.001) and highest in the C group. Bacterial challenge decreased bone metabolism markers in all groups (P < 0.01). Further, osteoclasts showed an augmented inflammatory response to P. gingivalis in the presence of hyperlipidemic PA levels as opposed to OA cultures, which responded similarly to controls. These findings indicate that the specific FA profile of diet rather than weight gain and obesity alone modulates bone metabolism and can therefore influence alveolar bone loss.
Optimism and Life Satisfaction as Determinants for Dental and General Health Behavior—Oral Health Habits Linked to Cardiovascular Risk Factors

Oral infections and cardiovascular diseases share common biological and behavioral risk factors. Psychosocial determinants could act as a link between general health behavior and dental health behavior. Our objective was to study optimism and life satisfaction as determinants of general and dental health behavior and to evaluate whether these are connected with cardiovascular risk factors and dental diseases. The 1966 Northern Finland Birth Cohort (N = 12,058) is a general population birth cohort. In a postal questionnaire, respondents (N = 8690) were asked about their health behavior and dental status. Cardiovascular risk factors were assessed in health examinations (N = 6033). Generalized linear regression models were used in analysis. The results showed that health orientation increases with strengthening life satisfaction and optimism. Dental health behavior and general health behavior were associated with both cardiovascular risk factors and self-reported dental diseases, which support the assumption that they share a common behavioral background.
How likely is environmental or patient cross-contamination of Chlamydia trachomatis DNA to lead to false positive results in patients attending our clinic?

Objectives Environmental contamination with DNA from Chlamydia trachomatis (CT) has previously been found in Genitourinary Medicine (GUM) clinics. There are no known cases of cross-contamination of clinical samples and no known nosocomial infections. We investigated whether diagnostic samples could become contaminated from the environment by running dummy sample and carrying out a patient-throughput analysis. A total of 29 748 patients attended clinics over a year. Of these, 2860 (9.6%) had a positive Chlamydia test result. Method (1) A run of dummy samples (60 urine samples and 10 swabs) were processed as normal clinic specimens. (2) Patient-throughput analysis: Patient numbers attending the GUM clinic on a given day was categorised as low, moderate or high. χ2 Tests were used to look for associations between categorical variables and Chlamydia test positivity. A Poisson regression model was fitted to look at the effect of the number of people in the clinic on the number of positive results in a given day. As some clinics were only run on certain days of the week, a sensitivity analysis was later performed with attendances at non-daily clinics removed. Results All dummy samples tested negative and we did not find evidence of an association between daily Chlamydia positivity and clinic attendance. Conclusions It is unlikely that environmental or cross-contamination of CT has lead to significant numbers of false positive results. Laboratories check for possible cross-contamination routinely. The extension of this simple routine practice to all clinical areas could provide quality assurance, improving confidence in the results in clinics.
Upper limb ischaemia - a single centre experience.

INTRODUCTION This paper reviews the epidemiology, aetiology and management of upper limb ischaemia in a series of 8 patients presenting to a tertiary referral centre over a 4-year period and the review of the pertinent literature. MATERIALS AND METHODS Details of patients admitted to Tan Tock Seng Hospital (TTSH) due to critical ischaemia of the upper limbs were obtained from admission summaries through the Computerised Patient Support System (CPSS) and operative notes through LOTUS. RESULTS There were 8 patients who presented with upper limb ischaemia excluding trauma and iatrogenic causes over the last 4 years. All patients underwent embolectomy. One patient had an amputation post-embolectomy. Our short-term results were encouraging with 7 patients who were well enough to be discharged. One patient had a massive brainstem stroke and was discharged home for comfort care at the request of the family. CONCLUSION Careful physical examination and history taking, prompt recognition of upper limb ischaemia and active approach to management in the form of embolectomy are crucial in obtaining a good outcome and reducing the risk of late disabling effects.
Direct Comparison of Placebo Effects on Clinical and Experimental Pain

Objectives:Placebo effects have been suggested to be more potent on clinical than experimental pain. However, this proposition is based on the comparison of the magnitude of placebo analgesia between studies using different methodologies or between different groups of subjects within the same study. The authors sought to provide a more direct test of this hypothesis using a within-subject design and to investigate the potential mediating effect of expectancy. Method:Sixteen patients with low back pain rated the intensity and the unpleasantness of their clinical pain and underwent two cold pressor tests, both before and after a saline injection presented either as a potent painkiller (placebo treatment) in one session or as an inactive substance in a control session. Results:The placebo treatment produced comparable increases in expected relief for clinical and experimental pain. However, ratings of pain intensity, pain unpleasantness, and perceived relief confirmed the larger placebo effect in low back pain than cold pressor pain. Retrospective ratings of perceived relief in low back pain generally showed the largest placebo effect compared with concurrent pain ratings. Furthermore, when the placebo session was performed after the control session, the placebo effect on low back pain was substantially reduced and observed only in perceived relief. Variations in expectation could not account for the large difference in placebo analgesia between clinical and experimental pain. Conclusions:The important reduction in placebo analgesia in low back pain after the single pre-exposure to the ineffective control treatment suggests the additional involvement of highly flexible mechanisms that may counteract the pro-analgesic effects of expectations.
Genotypes and molecular epidemiology of human astroviruses in hospitalized children with acute gastroenteritis in Rio de Janeiro, Brazil

Human astrovirus (HAstV) is an important pathogen that causes acute gastroenteritis mainly in children up to 5 years old worldwide. A total of 318 fecal samples were collected from January to December of 2004 from children with acute gastroenteritis hospitalized in Rio de Janeiro (RJ), Brazil. The samples were obtained from three public hospitals and were previously investigated for the presence of rotavirus (RV) and enteric adenovirus (AdV). The aim of this study was to determine the prevalence and molecular characterization of HAstV by reverse‐transcription polymerase chain reaction (RT‐PCR) and partial sequencing of open reading frame 2 (ORF2). HAstV infections were detected in 43 (14%) of the samples, of which 19 (6%) were mixed infections with HAstV and RV. HAstV were identified in all age groups, with 85% of the cases occurring in children younger than 2 years old. HAstV infections were more prevalent from March to May. Diarrhea, vomiting, and fever were the main clinical manifestations observed in these infections. Partial capsid sequencing of 35 HAstV strains characterized 25 (71%) as genotype 1 (HAstV‐1), 6 (17%) HAstV‐2, 3 (9%) HAstV‐8, and 1 (3%) HAstV‐4. Based on the genetic variability, three lineages were observed between RJ HAstV‐1 strains. This study demonstrated the importance of HAstV infections causing acute infantile gastroenteritis in hospitalized children in RJ, Brazil. Molecular epidemiological surveillance is helpful for designing prevention strategies of HAstV transmission. J. Med. Virol. 79:939–944, 2007. © 2007 Wiley‐Liss, Inc.