Miracle-dz/UniFAR-FTUs huggingface dataset:
Time course of recovery of endothelial cell surface thrombin receptor (PAR-1) expression.

We studied dynamics of cell surface expression of proteolytically activated thrombin receptor (PAR-1) in human pulmonary artery endothelial cells (HPAEC). PAR-1 activation was measured by changes in cytosolic calcium concentration ([Ca2+]i) and HPAEC retraction response (determined by real-time transendothelial monolayer electrical resistance). [Ca2+]iincrease in response to thrombin was abolished by preexposure to 25 nM thrombin for >60 min, indicating PAR-1 desensitization, but preexposure to 25 nM thrombin for only 30 min or to 10 nM thrombin for up to 2 h did not desensitize PAR-1. Exposure to 10 or 25 nM thrombin decreased monolayer electrical resistance 40-60%. Cells preexposed to 10 nM thrombin, but not those preexposed to 25 nM thrombin, remained responsive to thrombin 3 h later. Loss of cell retractility was coupled to decreased cell surface PAR-1 expression as determined by immunofluorescence. Cell surface PAR-1 disappeared upon short-term (30 min) thrombin exposure but reappeared within 90 min after incubation in thrombin-free medium. Exposure to 25 nM thrombin for >60 min prevented rapid cycloheximide-insensitive PAR-1 reappearance. Cycloheximide-sensitive recovery of cell surface PAR-1 expression required 18 h. Therefore, both duration and concentration of thrombin exposure regulate the time course of recovery of HPAEC surface PAR-1 expression. The results support the hypothesis that initial recovery of PAR-1 surface expression in endothelial cells results from a rapidly mobilizable PAR-1 pool, whereas delayed recovery results from de novo PAR-1 synthesis. We conclude that thrombin itself regulates endothelial cell surface PAR-1 expression and that decreased surface expression interferes with thrombin-induced endothelial cell activation responses.
More acidic dietary acid-base load is associated with reduced calcaneal broadband ultrasound attenuation in women but not in men: results from the EPIC-Norfolk cohort study.

BACKGROUND Dietary patterns that promote mild metabolic acidosis may have a negative effect on bone density. OBJECTIVE We investigated the relation between a measure of dietary acid-base load, potential renal acid load (PRAL), and calcaneal broadband ultrasound attenuation (BUA) after adjustment for confounders and also compared the results with different estimates of acid-base load. DESIGN A cross-sectional study was conducted in 14 563 men and women aged 42-82 y living in Norfolk, United Kingdom, in which measures of calcaneal BUA and dietary PRAL were estimated by using the European Prospective Investigation into Cancer and Nutrition Norfolk (EPIC-Norfolk) food-frequency questionnaire. RESULTS A more acidic dietary intake (high PRAL) was significantly associated with lower calcaneal BUA in women but not in men; there was a difference of approximately 2% in BUA between the highest and lowest quintiles of PRAL, independent of age, body mass index, smoking habit, physical activity, diagnosed osteoporosis, and history of fracture, and (in women) hormone replacement therapy. No relation was observed between history of fracture or incident fracture and PRAL. Those with the greatest PRAL had higher intakes of meat, fish, eggs, and cereal and cereal products and lower intakes of fruit and vegetables, tea, and coffee. CONCLUSION PRAL was inversely associated with bone ultrasound measures in women, but the magnitude of the association was relatively small compared with other known risk factors. Further longitudinal studies are required to establish whether, in the long term, these small effects are important in overall fracture risk in populations.
Endogenous attention improves perception in amblyopic macaques

Amblyopia, a developmental disorder of vision, affects many aspects of spatial vision as well as motion perception and some cognitive skills. Current models of amblyopic vision based on known neurophysiological deficiencies have yet to provide an understanding of the wide range of amblyopic perceptual losses. Visual spatial attention is known to enhance performance in a variety of detection and discrimination tasks in visually typical humans and nonhuman primates. We investigated whether and how voluntary spatial attention affected psychophysical performance in amblyopic macaques. Full-contrast response functions for motion direction discrimination were measured for each eye of six monkeys: five amblyopic and one control. We assessed whether the effect of a valid spatial cue on performance corresponded to a change in contrast gain, a leftward shift of the function, or response gain, an upward scaling of the function. Our results showed that macaque amblyopes benefit from a valid spatial cue. Performance with amblyopic eyes viewing showed enhancement of both contrast and response gain whereas fellow and control eyes' performance showed only contrast gain. Reaction time analysis showed no speed accuracy trade-off in any case. The valid spatial cue improved contrast sensitivity for the amblyopic eye, effectively eliminating the amblyopic contrast sensitivity deficit. These results suggest that engaging endogenous spatial attention may confer substantial benefit to amblyopic vision.
Multiple micronutrient supplementation during pregnancy does not lead to greater infant birth size than does iron-only supplementation: a randomized controlled trial in a semirural community in Mexico.

BACKGROUND Little is known about the benefits of prenatal multivitamin and mineral supplements in reducing low birth weight. OBJECTIVE We conducted a randomized, double-blind clinical trial in semirural Mexico to compare the effects of multiple micronutrient (MM) supplements with those of iron supplements during pregnancy on birth size. DESIGN Pregnant women (n = 873) were recruited before 13 wk of gestation and received supplements 6 d/wk at home, as well as routine antenatal care, until delivery. Both supplements contained 60 mg Fe, but the MM group also received 1-1.5 times the recommended dietary allowances of several micronutrients. RESULTS At recruitment, the women in the 2 groups were not significantly different in age, parity, economic status, height, or hemoglobin concentration but differed significantly in marital status (4.6% and 2.0% of women in the MM and iron-only groups, respectively, were single mothers) and mean (+/- SD) body mass index (in kg/m(2); 24.6 +/- 4.3 and 23.8 +/- 3.9 in the iron-only and MM groups, respectively). Losses to follow-up (25%) and compliance (95%) did not differ significantly between the groups. In intent-to-treat analyses (MM group: n = 323; iron-only group: n = 322), mean (+/- SD) birth weight (2.981 +/- 0.391 and 2.977 +/- 0.393 kg in the MM and iron-only groups, respectively) and birth length (48.61 +/- 1.82 and 48.66 +/- 1.83 cm in the MM and iron-only groups, respectively) did not differ significantly between the groups. CONCLUSION These findings suggest that MM supplementation during pregnancy does not lead to greater infant birth size than does iron-only supplementation.
Dynamic heart rate variability: a tool for exploring sympathovagal balance continuously during sleep in men.

We have recently demonstrated that the overnight profiles of cardiac interbeat autocorrelation coefficient of R-R intervals ( r RR) calculated at 1-min intervals are related to the changes in sleep electroencephalographic (EEG) mean frequency, which reflect depth of sleep. Other quantitative measures of the Poincaré plots, i.e., the standard deviation of normal R-R intervals (SDNN) and the root mean square difference among successive R-R normal intervals (RMSSD), are commonly used to evaluate heart rate variability. The present study was designed to compare the nocturnal profiles of r RR, SDNN, and RMSSD with the R-R spectral power components: high-frequency (HF) power, reflecting parasympathetic activity; low-frequency (LF) power, reflecting a predominance of sympathetic activity with a parasympathetic component; and the LF-to-HF ratio (LF/HF), regarded as an index of sympathovagal balance. r RR, SDNN, RMSSD, and the spectral power components were calculated every 5 min during sleep in 15 healthy subjects. The overnight profiles of r RR and LF/HF showed coordinate variations with highly significant correlation coefficients ( P < 0.001 in all subjects). SDNN correlated with LF power ( P < 0.001), and RMSSD correlated with HF power ( P < 0.001). The overnight profiles of r RR and EEG mean frequency were found to be closely related with highly cross-correlated coefficients ( P < 0.001). SDNN and EEG mean frequency were also highly cross correlated ( P < 0.001 in all subjects but 1). No systematic relationship was found between RMSSD and EEG mean frequency. In conclusion, r RR appears to be a new tool for evaluating the dynamic beat-to-beat interval behavior and the sympathovagal balance continuously during sleep. This nonlinear method may provide new insight into autonomic disorders.
A novel method for analysis of nuclear receptor function at natural promoters: peroxisome proliferator-activated receptor gamma agonist actions on aP2 gene expression detected using branched DNA messenger RNA quantitation.

Peroxisome proliferator-activated receptor-gamma (PPARgamma), a member of the nuclear hormone receptor superfamily, plays an essential role in the mediation of the actions of antidiabetic drugs known as thiazolidinediones (TZDs). PPARgamma activates many target genes involved in lipid anabolism including the adipocyte fatty acid binding protein (aP2). In this study, induction of aP2 gene expression by PPARgamma agonists was examined in both cultured cells and diabetic mice using branched DNA (bDNA)-mediated mRNA quantitation. bDNA technology allows for the direct measurement of a particular mRNA directly within cellular lysate using a 96-well plate format in a time frame comparable to a reporter gene assay. In cultured human subcutaneous preadipocytes, the TZDs, troglitazone and BRL-49653, both rapidly induced aP2 mRNA as detected with the bDNA method. In these cells, the effect of BRL-49653 on aP2 mRNA levels was detectable as early as 30 min after treatment (47% increase) and was maximal after 24 h of treatment (12-fold increase). The effects of troglitazone on aP2 mRNA induction were similar to those of BRL-49653 except that the maximal level of induction was consistently lower (e.g. 24 h treatment = 4-fold increase). Dose-response relationships for both of the TZDs were also determined using the 24-h treatment time point. EC50s for both BRL-49653 and troglitazone were estimated to be 80 nM and 690 nM, respectively. A natural PPARgamma ligand, 15-deoxy-delta12,14-PGJ2, was also active in this assay with a maximal induction of aP2 mRNA of approximately 5-fold when tested at 1 microM. Since the PPARgamma:retinoid X receptor (RXR) heterodimer has been characterized as a permissive heterodimer with respect to RXR ligands, the ability of 9-cis-retinoic acid (9-cis-RA) to induce aP2 mRNA was examined. Although 9-cis-RA had very low efficacy (2-fold induction), the maximal effect was reached at 100 nM. No synergism or additivity in aP2 mRNA induction was detected when 9-cis-RA was included with either of the TZDs used in this study. Significant induction of aP2 mRNA in bone marrow of db/db mice treated with either troglitazone or BRL-49653 was also detected, indicating that the bDNA assay may be a simple method to monitor nuclear receptor target gene induction in vivo.
Angiotensin II acts at AT1receptors in the nucleus of the solitary tract to attenuate the baroreceptor reflex.

The object of the current study was to determine if ANG II acts at type 1 (AT1) or type 2 (AT2) receptors in the nucleus of the solitary tract (NTS) to reduce baroreceptor reflex control of renal sympathetic nerve activity (RSNA) and heart rate (HR). Experiments were carried out in urethan-anesthetized Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR). Reflex changes in RSNA and HR were elicited by intravenous infusion of either phenylephrine or sodium nitroprusside before and after bilateral microinjection of CV-11974 (AT1 receptor antagonist, 10 pmol), PD-123319 (AT2 receptor antagonist, 100 pmol), or artificial cerebrospinal fluid (aCSF, 50 nl) in the NTS. Mean arterial pressure (MAP)-RSNA and MAP-HR data were fit to logistic functions to analyze the baroreceptor reflex. Baroreceptor reflex sensitivities for RSNA and HR were attenuated in SHR compared with those in WKY rats. Bilateral injection of CV-11974, PD-123319, or aCSF in the NTS of either strain had no effect on baseline arterial pressure, HR, or RSNA. However, CV-11974 injected in the NTS increased significantly ( P < 0.01) the sensitivities for baroreceptor reflex control of RSNA and HR in SHR and WKY rats. Neither PD-123319 nor aCSF altered baroreceptor reflex control of RSNA and HR in either SHR or WKY rats. These results demonstrate that endogenous ANG II acts at AT1 receptors of the NTS to attenuate the baroreceptor reflex in SHR as well as in WKY rats.
Association of Alcohol With Mortality After Traumatic Brain Injury

Although alcohol exposure results in reduced mortality after traumatic brain injury (TBI) in animal models, clinical trials based on proposed mechanisms have been disappointing and have reported conflicting results. Methodological issues common to many of these clinical studies may have contributed to the spurious results. Our objective was to evaluate the association between blood alcohol concentration (BAC) and in-hospital mortality after TBI, and overcome methodological problems of prior studies. We conducted a retrospective cohort study on individuals treated for isolated TBI (n = 1,084) at the R Adams Cowley Shock Trauma Center (Baltimore, Maryland) from 1997 to 2012. We excluded individuals with injury to other body regions and examined multiple cutpoints of BAC. Our primary outcome was in-hospital mortality. In adjusted logistic regression models, the upper level of each blood alcohol categorization from 0.10 g/dL (odds ratio = 0.63, 95% confidence interval: 0.40, 0.97) through 0.30 g/dL (odds ratio = 0.25, 95% confidence interval: 0.08, 0.84) was associated with reduced risk of mortality after TBI compared with individuals with undetectable BAC. In sensitivity analyses among individuals without penetrating brain injuries (95% firearm-related) (n = 899), the protective association was eliminated. This study provides evidence that the observed protective association between BAC and in-hospital mortality after TBI resulted from bias introduced by inclusion of penetrating injuries.
Synthetic flavonoids cross the placenta in the rat and are found in fetal brain.

The synthetic flavonoid EMD-49209 is a potent inhibitor of the in vivo and in vitro binding of thyroxine (T4) to transthyretin (TTR). We studied the distribution of125I-labeled EMD-49209 in maternal tissues, intestinal contents, and fetal tissues in rats that were 20 days pregnant (from 1 to 24 h after intravenous injection). The percent dose of EMD decreased quickly with time. In maternal brain no radioactive flavonoid could be detected. EMD was excreted very rapidly from the intestines. In the fetal compartment the percent dose of EMD increased with time; after 24 h it contained 17% of the EMD. The flavonoid was found in all fetal tissues investigated and also in the fetal brain. Because TTR concentrations are high in the fetal rat, especially in the brain, the transfer of flavonoid to the fetal brain might be linked to TTR expression. The presence of flavonoid in the fetal brain raises the possibility of an essential interference of flavonoids with the availablity of T4 in the fetal compartment.
Control of hepatic insulin-like growth factor II gene expression by thyroid hormones in fetal sheep near term.

The effects of thyroid hormones on hepatic insulin-like growth factor (IGF) II gene expression and their interaction with cortisol in the ontogenic control of this gene were investigated in fetal sheep during late gestation (term 145 ± 2 days) and after experimental manipulation of fetal plasma hormone concentrations. In intact fetuses, a significant decrease in hepatic IGF-II mRNA abundance was observed between 127-130 and 142-145 days of gestation, which coincided with the normal prepartum rise in plasma cortisol and triiodothyronine (T3) concentrations. This ontogenic decline in hepatic IGF-II gene expression was abolished in fetuses in which the prepartum rise in plasma T3, but not cortisol, was prevented by fetal thyroidectomy. At 127-130 days, downregulation of hepatic IGF-II mRNA abundance was induced prematurely in intact fetuses by an infusion of cortisol for 5 days (2-3 mg ⋅ kg-1 ⋅ day-1iv). Plasma concentrations of cortisol and T3 in the cortisol-infused intact fetuses were increased to values seen close to term. Similar findings were observed in thyroidectomized fetuses, in which, despite thyroidectomy, cortisol infusion significantly increased plasma T3 concentrations and caused a premature decrease in hepatic IGF-II mRNA levels. However, in intact fetuses at 127-130 days, the increasing of T3 concentrations alone by exogenous T3 infusion (8-12 μg ⋅ kg-1 ⋅ day-1iv for 5 days) had no effect on hepatic IGF-II mRNA levels. Overall, a decrease in hepatic IGF-II mRNA abundance was only observed in fetuses in which there were concurrent increases in plasma cortisol and T3 concentrations. When observations from all fetuses were considered, irrespective of gestational age or treatment, hepatic IGF-II mRNA levels were negatively correlated with plasma cortisol and T3 but not thyroxine concentrations. Partial correlation analysis of hepatic IGF-II, cortisol, and T3 values showed that the plasma concentration of cortisol in the fetus had the predominant effect on hepatic IGF-II mRNA abundance. These findings show that T3 may mediate, in part, the maturational effects of cortisol on hepatic IGF-II gene expression but that it is ineffective without a concomitant rise in fetal plasma cortisol. Hence, increased concentrations of both cortisol and T3 appear necessary to induce downregulation of hepatic IGF-II mRNA abundance in fetal sheep close to term.
Effects of multiple simulated birth traumas on urethral continence function in rats.

Multiple vaginal parities have been reported to be an important risk factor for stress urinary incontinence (SUI). Simulated birth trauma with single vaginal distention (VD) has been used to induce the SUI condition in animals; however, the effect of multiple simulated birth traumas on the urethral continence function has not been well characterized. Therefore, we examined the effects of multiple VDs on urethral functions in vivo and the changes in gene expressions of several molecules in the urethra using female SD rats, which were divided into three groups; sham, VD-1 (single VD), and VD-3 groups (3 times of VDs every 2 wk). Two weeks after the final VD, leak point pressure (LPP) and urethral responses during sneezing were evaluated. Also, changes in mRNA levels of urethral molecules were quantified with RT-PCR. The VD-1 group did not show any change in LPP with only a tendency of decrease in amplitudes of the urethral responses during sneezing (A-URS); however, the VD-3 group showed a significant decrease in LPP and urethral responses such as baseline urethral pressure and A-URS accompanied with SUI episodes during sneezing. Nicotinic receptor subtypes and transforming growth factor (TGF)-β1 were significantly increased in both VD-1 and VD-3 groups while TNF receptor (TNFR)-1, IL-6, collagens, and matrix metalloproteinases-9 were significantly increased only in the VD-3 group. These data indicate that rats with multiple simulated birth traumas exhibit profound impairment of the urethral continence function and that these functional changes are associated with those in cytokines, extracellular matrix molecules, and nicotinic receptor subtypes in the urethra.
Effects of acidosis on Ca2+sensitivity of contractile elements in intact ferret myocardium.

We investigated the effects of acidosis on the intracellular Ca2+ concentration ([Ca2+]i) and contractile properties of intact mammalian cardiac muscle during tetanic and twitch contractions. Aequorin was injected into ferret papillary muscles, and the [Ca2+]iand tension were simultaneously measured. Acidosis was attained by increasing the CO2 concentration in the bicarbonate (20 mM)-buffered Tyrode solution from 5% (pH 7.35, control) to 15% (pH 6.89, acidosis). Tetanic contraction was produced by repetitive stimulation of the preparation following treatment with 5 μM ryanodine. The relationship between [Ca2+]iand tension was measured 6 s after the onset of the stimulation and was fitted using the Hill equation. Acidosis decreased the maximal tension to 81 ± 2% of the control and shifted the [Ca2+]i-tension relationship to the right by 0.18 ± 0.01 pCa units. During twitch contraction, a quick shortening of muscle length from the length at which developed tension became maximal ( L max) to 92% L maxproduced a transient change in the [Ca2+]i(extra Ca2+). The magnitude of the extra Ca2+ was dependent on the [Ca2+]iimmediately before the length change, suggesting that the extra Ca2+ is related to the amount of troponin-Ca complex. Acidosis decreased the normalized extra Ca2+ to [Ca2+]iimmediately before the length change, which indicates that the amount of Ca2+ bound to troponin C is less when [Ca2+]iis the same as in the control. The decrease in the Ca2+ binding to troponin C explains the decrease in tetanic and twitch contraction, and mechanical stress applied to the preparation induced less [Ca2+]ichange in acidosis.
Systematic Complex Haploinsufficiency-Based Genetic Analysis of Candida albicans Transcription Factors: Tools and Applications to Virulence-Associated Phenotypes

Genetic interaction analysis is a powerful approach to the study of complex biological processes that are dependent on multiple genes. Because of the largely diploid nature of the human fungal pathogen Candida albicans, genetic interaction analysis has been limited to a small number of large-scale screens and a handful for gene-by-gene studies. Complex haploinsufficiency, which occurs when a strain containing two heterozygous mutations at distinct loci shows a phenotype that is distinct from either of the corresponding single heterozygous mutants, is an expedient approach to genetic interactions analysis in diploid organisms. Here, we describe the construction of a barcoded-library of 133 heterozygous TF deletion mutants and deletion cassettes for designed to facilitate complex haploinsufficiency-based genetic interaction studies of the TF networks in C. albicans. We have characterized the phenotypes of these heterozygous mutants under a broad range of in vitro conditions using both agar-plate and pooled signature tag-based assays. Consistent with previous studies, haploinsufficiency is relative uncommon. In contrast, a set of 12 TFs enriched in mutants with a role in adhesion were found to have altered competitive fitness at early time points in a murine model of disseminated candidiasis. Finally, we characterized the genetic interactions of a set of biofilm related TFs in the first two steps of biofilm formation, adherence and filamentation of adherent cells. The genetic interaction networks at each stage of biofilm formation are significantly different indicating that the network is not static but dynamic.