Miracle-dz/UniFAR-FTUs huggingface dataset:
Determining the Requirements for Cooperative DNA Binding by Swi5p and Pho2p (Grf10p/Bas2p) at the HO Promoter (*)

SWI5 encodes a zinc finger DNA binding protein required for the transcription of the Saccharomyces cerevisiae HO gene, and PHO2 encodes a homeodomain DNA binding protein. In vitro biochemical studies using purified Swi5p and Pho2p proteins have demonstrated that Swi5p and Pho2p bind cooperatively to the HO promoter. In this report we investigate the regions of the Swi5p and Pho2p proteins required for cooperative DNA binding. The analysis of each protein gives a similar result: the zinc finger or homeodomain DNA binding domains are each sufficient for in vitro DNA binding, but additional regions of each protein are required for cooperative DNA binding. In vitro and in vivo experiments were conducted with promoters with altered spacing between the Pho2p and Swi5p binding sites. Mutations that increased the distance between the two binding sites had minimal effects on either in vitro cooperative DNA binding or in vivo upstream activating sequence activity. These observations suggest that the interaction domains of Swi5p and Pho2p are flexible and can tolerate an increase in distance between the two binding sites. The mechanism of the cooperative DNA binding by Swi5p and Pho2p is discussed.
Responses to submaximal treadmill exercise and training in the horse: changes in haematology, arterial blood gas and acid base measurements, plasma biochemical values and heart rate

Four standardbred horses with subcutaneously relocated carotid arteries were given a seven week training programme of treadmill exercise at a gradient of 19 per cent in order to assess if there were any effects of exercise and training on haematology, arterial blood gas and acid base measurements, plasma biochemistry and heart rate. The exercise consisted of one minute walking at 110 metres/minute followed by five minutes trotting at 200 metres/minute, twice daily in the first week. The period of trotting exercise was increased by one minute per week so that by the seventh week the horses were being given 12 minutes trotting twice daily. Before training commenced venous blood samples, for complete blood counts and plasma biochemistry, and arterial samples, for blood gas, acid base and lactate measurements, were taken at rest, after five minutes and 15 minutes of treadmill exercise (200 metres/minute) and 30 minutes and 60 minutes after completing the exercise. Heart rate was measured by telemetric electrocardiogram at similar intervals. This exercise test and blood collection were repeated after one, three, five and seven weeks of training. The only significant changes were a decrease in exercise lactate with training, increases in exercise and recovery total protein. The haematological response to treadmill exercise included an increase in certain red cell parametes and a leucocytosis which was caused by both a neutrophilia and a lymphocytosis. These effects had largely disappeared by 30 minutes after exercise and all values had returned to resting values by one hour after exercise. (ABSTRACT TRUNCATED AT 250 WORDS)
[Effect of early cleavage of embryos after intracytoplasmic sperm injection (ICSI) into oocytes on results of fertilization in vitro (IVF) and embryo transfer (ET)].

OBJECTIVE To evaluate the influence of early cleavage in embryos obtained using ICSI on the success rate of IVF-ET. DESIGN A prospective study. SETTING Centre of Assisted Reproduction, Department of Obstetrics and Gynecology, Medical Faculty, Palacký University, Olomouc. METHODS The rate of the first cleavage of embryos was checked between 23 to 26 hours after ICSI in 115 cycles IVF-ET. "Early Cleavage" (EC) embryos which were already cleaved were cultivated separately from the "slow" (No Early Cleavage, NEC) embryos. After 3 days cultivation, no more than three 6- to 8-cell embryos were transferred. Only EC embryos were transferred in 43 cycles and only NEC embryos were transferred also in 43 cycles. The remaining 29 cycles, where both EC and NEC embryos had to be transferred, were not evaluated. In both groups the following entry parameters were compared: age of the patient, number of fertilized oocytes, fertilization rate (FR), number of frozen and evaluated embryos, number of EC embryos recorded and number of transferred embryos. The outcome parameters compared were: pregnancy rate (PR), number of gestational sacs with heart beats per number of transferred embryos (implantation rate, IR), abortions (AB) and the frequency of multiple pregnancy. Statistical evaluation was performed by t-test, chi 2 test and by Sheffe analysis. RESULTS EC embryos were found in 57% of cycles (n = 66) after ICSI. Early cleavage was observed in 198 of 563 embryos (35%). Among entry parameters the only statistically significant differences (P < 0.05) were found in the number of retrieved oocytes as well as in the ratio of frozen embryos which were higher in the cycles with the transfer of only EC embryos. FR was the same (74%) in both types of transfer. Outcome parameters in the transfer of only EC versus NEC embryos were higher in PR (40% vs. 33%), about equal in IR (27% vs. 25%) and lower in AB (2% vs. 5%). Moreover, multiple pregnancies were less frequent in the transfer of only EC embryos: 65% of singleton pregnancy, 35% of twins and no triplet versus 36%, 57% and 7%, resp. , in the transfer of only NEC embryos. The differences in outcome parameters are not statistically significant (P > 0.05) because of small numbers. CONCLUSION The speed of the first cell cleavage is a useful additional criterion for better evaluation of quality and viability of embryos.
How should we estimate driving pressure to measure interrupter resistance in children?

Interrupter resistance (Rint) is a widely used measure of airway caliber, but concerns remain about repeatability and sensitivity. Some Rint variability may derive from the linear back‐extrapolation algorithm (LBE 30/70) usually used to estimate driving pressure. To investigate whether other methods of estimating driving pressure could improve repeatability and sensitivity, we studied 39 children with asthma. Two measurements of Rint—each the median of 10 interruptions—were made 5 min apart, and 14 children had a third measurement after bronchodilator (RintBD). Mouth pressure transients were analyzed using several algorithms, to compare the magnitude, repeatability, and sensitivity to bronchodilator change of Rint values yielded. Algorithms taking driving pressure from later in the transient, predictably, yielded higher values of Rint than those which back‐extrapolated to time of valve closure. Algorithms which did not rely on back‐extrapolation, including mean oscillation pressure (MOP) and mean plateau pressure (MP 30/70) had better repeatability. Sensitivity to detect change, calculated as ratio of bronchodilator response to repeatability coefficient (ΔRint/CR), was also better for non‐extrapolating algorithms: MP 30/70 1.67, LBE 30/70 1.28 (P = 0.0004). Measuring Rint using techniques other than conventional back‐extrapolation may give more consistent and clinically useful results, and these approaches merit further exploration. Pediatr Pulmonol. 2007; 42:757–763. © 2007 Wiley‐Liss, Inc.
The Canadian Network for Mood and Anxiety Treatments (CANMAT) task force recommendations for the management of patients with mood disorders and comorbid metabolic disorders.

BACKGROUND One goal of the Canadian Network for Mood and Anxiety Treatments (CANMAT) is to develop evidence-based and best practice educational programs and recommendations. Our group conducted a comprehensive literature review to provide evidence-based recommendations for treating metabolic comorbidity in individuals with major depressive disorder (MDD) and bipolar disorder (BD). METHODS We searched PubMed for all English-language articles published January 1966 to November 2010 using BD and MDD cross-referenced with metabolic syndrome, obesity, diabetes mellitus, hypertension, and dyslipidemia. That search was augmented by a review of articles reporting outcomes of an intervention targeting components of metabolic syndrome in individuals with MDD or BD. RESULTS Consensus exists for the recommendation that individuals with MDD and BD should be routinely screened for risk factors that increase risk for metabolic syndrome. For excess weight, the best-studied pharmacologic approaches are metformin and topiramate, with emerging evidence for liraglutide and modafinil. For binge eating disorder, the best evidence in mood disorders was for cognitive-behavioral therapy as well as topiramate, zonisamide, and in select cases selective serotonin reuptake inhibitors. For dysglycemia, dyslipidemia, and hypertension, evidence supports cognitive-behavioral interventions and anti-diabetic, antilipidemic, and antihypertensive treatments. CONCLUSIONS Comprehensive care of individuals with mood disorders should include routine evaluation of the risk and presence of metabolic syndrome and its components. Systematic evaluation of preventative and targeted treatments of metabolic syndrome in mood disorder populations is insufficient.
Mandibular two-implant overdentures: prosthodontic maintenance using different loading protocols and attachment systems.

PURPOSE The aim of this research was to determine the long-term prosthodontic maintenance requirements of mandibular two-implant overdentures using different loading protocols and attachment systems. MATERIALS AND METHODS A total of 106 participants were allocated randomly to one of four different implant systems (Steri-Oss, Southern, Straumann, or Branemark). Three different loading protocols (2, 6, and 12 weeks) were used with six different ball abutment patrices and their respective matrices (Steri-Oss rubber, Straumann gold, Straumann titanium, Branemark gold, Southern plastic, and Southern gold/platinum). Prosthodontic maintenance events were documented prospectively from baseline until the 8-year recall according to predefined categories. RESULTS After 6 years, 90 participants attended recall and, thereafter, 68 participants were followed for 8 years. No significant differences were found between the number of prosthodontic maintenance events and the loading protocol used. Steri-Oss rubber matrices had the highest mean number of maintenance events at 32.2 ± 14.5 events, followed by the Branemark gold matrices at 28.8 ± 12.6 events. The Southern plastic matrices had a significantly lower mean number of maintenance events (8.7 ± 4.2) when compared with all other groups. Over a 6-year period, the matrices with the best longevity were Straumann gold at 3.9 ± 2.1 years. Straumann gold matrices also lasted significantly longer than all other matrices (P < .05). Southern gold/platinum, Branemark gold, and Southern plastic matrices all lasted significantly longer than the Straumann titanium and Steri-Oss matrices (P < .05). The mean time to reline for overdentures was 3.37 ± 2.06 years; remaking of overdentures peaked by year 7, with a mean time to remake of 5.81 ± 2.04 years. CONCLUSION Early loading protocols do not influence long-term prosthodontic maintenance requirements of unsplinted mandibular two-implant overdentures. By contrast, attachment systems do influence prosthodontic maintenance, particularly with regard to the type of matrices used.
Improving handwriting through kinaesthetic sensitivity practice.

Children in their first two years of formal education (kindergarten and year one) were assessed for handwriting performance. Those with poor handwriting were given practice on one of three tasks: kinaesthetic acuity, kinaesthetic memory and perception or a handwriting task. Both forms of kinaesthetic sensitivity practice produced significant improvement in handwriting performance by the year one children, while handwriting practice did not. The younger children did not demonstrate as clear cut gains as the older. These results support the findings of previous research which indicates the importance of kinaesthetic ability for performance of complex motor tasks.
Validation of a Single Concentration Methacholine Inhalation Provocation Test (SCIPT) in Children

A new method to assess bronchial hyperresponsiveness (BHR) using a single concentration methacholine has already been validated in adults with asthma. Because the geometrical dimensions of the airways in children are different, the results from studies in adults cannot be extrapolated to children. In this study, we validated the single concentration methacholine inhalation provocation test (SCIPT) in children. Twenty-two children performed three methacholine inhalation challenge tests in random order. Two challenges were performed according to the SCIPT: doubling doses (0.03–1.8 mg; maximal cumulated dose 3.6 mg) were administered with an Aerosol Provocation System (Masterscope, Jaeger). The third challenge was performed according to a standard dosimeter method (SDM): doubling doses (0.002–1.8 mg; maximal cumulative dose 3.5 mg) were administered with a DeVillbiss 646 nebulizer. The degree of BHR is expressed as a PD20. A difference of < 1.5 dose step was assumed to be due to intraindividual variation. We found an intraclass correlation of 0.91 between both tests according to the SCIPT and of 0.80 between the SCIPT and SDM. We found, according to the method of Bland and Altman, good agreement when comparing these two challenge tests. The single concentration inhalation provocation test is reproducible and shows good agreement with a standard dosimeter method to test bronchial responsiveness in children.
Photosensitizing properties of chlorins in solution and in membrane-mimicking systems

The photosensitizing properties of three chlorins, meso -tetra(3-hydroxyphenyl)chlorin ( m -THPC), chlorin e6 (Ce6) and meso -tetraphenylchlorin substituted by two adjacent sulfonated groups (TPCS_2a) are compared in solution and when incorporated in dioleoyl-sn-phosphatidylcholine (DOPC) liposomes. In solution, the three chlorins possess a similar efficacy to generate singlet oxygen (quantum yield ∼0.65). The formation of conjugated dienes was used to determine their ability to induce the peroxidation of methyl linoleate as a target of singlet oxygen. In ethanol solution, the apparent quantum yield for this process is the same for the three chlorins and its value agrees with that expected from the known rates for the decay of singlet oxygen and its reaction with methyl linoleate. When incorporated in liposomes, the order of efficacy is m -THPC > TPCS_2a > Ce6. This order is tentatively assigned to the relative embedment of the photosensitizer within the lipidic bilayer, TPCS_2a and Ce6 being anchored by their negative chains nearer to the water–lipid interface. The photoinduced permeation of the lipidic bilayer by these chlorins was investigated by measuring the release of carboxyfluorescein entrapped into DOPC liposomes. The charged chlorins, in particular TPCS_2a, are the most efficient, a result discussed in relation with the technology of photochemical internalization, PCI.
Human apolipoprotein E. Role of arginine 61 in mediating the lipoprotein preferences of the E3 and E4 isoforms.

Abstract Human apolipoprotein (apo) E4 (arginine at residue 112) preferentially associates with very low density lipoproteins (VLDL), and apoE3 (cysteine at 112) associates with high density lipoproteins. It has been postulated that the amino-terminal domain, which contains residue 112, influences the lipoprotein preference by interacting with the carboxyl-terminal domain, which contains the lipid-binding region. To delineate the region in the carboxyl-terminal domain mediating lipoprotein binding and involved in isoform preference, we produced truncated apoE3 and apoE4 variants (terminating at residues 251, 260, 266, or 272) in Escherichia coli and assessed them for lipoprotein association. This analysis suggested that residues 260-272 contain important determinants for complete lipoprotein association and isoform preferences. To determine whether positive charge at residue 112 was an absolute requirement for the apoE4 VLDL preference, we compared the distributions of rabbit apoE (equivalent to apoE3, with cysteine at a position corresponding to 112), canine apoE (arginine at the corresponding site), and cysteamine-treated rabbit apoE (cysteine converted to a positively charged residue). Surprisingly, all distributed like human apoE3, suggesting that positive charge at a position corresponding to 112 was not directly responsible for the isoform preference and that other residues in the amino-terminal domain were involved. To determine which residues were involved, the structure of the apoE4 22-kDa fragment (the amino-terminal two-thirds of the molecule) was determined to 2.5 A by x-ray crystallography. Compared with the known four-helix bundle structure of apoE3, the only significant differences in the apoE4 structure were that glutamic acid 109 formed a salt bridge with arginine 112 and that the arginine 61 side chain was displaced to a new position. Site-directed mutagenesis of glutamic acid 109 in apoE3 and arginine 61 in apoE4 demonstrated that the position of the arginine 61 side chain in apoE4 was critical in determining apoE4 lipoprotein distribution, suggesting that arginine 61 interacted with the carboxyl-terminal domain to direct binding to VLDL.
Radiation exposure during midfacial imaging using 4- and 16-slice computed tomography, cone beam computed tomography systems and conventional radiography.

OBJECTIVES Radiation doses were determined to balance risks against usefulness of the different modalities available for the imaging of the facial skeleton. METHODS An Alderson Rando Phantom, armed with lithium fluoride thermoluminescent dosemeters (TLDs) was exposed using a set of four conventional radiographs (orbital view, modified Waters view, orthopantomography, skull posterior--anterior 0 degrees ), two different cone beam computed tomography (CBCT) (NewTom 9000 and Siremobil Iso-C3D), and multislice computed tomography (CT) modalities (Somatom VolumeZoom and Somatom Sensation 16). TLDs from 14 well defined anatomical sites lying within the primary beam as well as the TLD corresponding to the thyroid gland were evaluated. RESULTS Multislice CT showed the highest exposure values. Exposure levels of the CBCT systems lay between CT and conventional radiography. Dose measurement for the 16-slice CT revealed nearly the same radiation exposure as the 4-slice system when adapted examination protocols were used. CONCLUSIONS Selection of the most appropriate imaging modality should be performed in view of the delivered doses, required image quality and information and the clinical circumstances.
Pioglitazone reduces inflammation through inhibition of NF-κB in polymicrobial sepsis

The insulin sensitizing thiazolidinedione drugs, rosiglitazone and pioglitazone are specific peroxisome proliferator-activated receptor-gamma agonists and reduce pro-inflammatory responses in patients with type 2 diabetes and coronary artery disease, and may be beneficial in sepsis. Sepsis was induced in 8–10-wk-old C57BL/6 mice by cecal ligation and puncture (CLP) with a 22 -g double puncture technique. Mice received an i.p. injection of vehicle (DMSO:PBS) or pioglitazone (20 mg/kg) at 1 h and 6 h after CLP, and were sacrificed at various time points. In sepsis, vehicle-treated mice had hypoglycemia, increased lung injury and increased lung neutrophil infiltration. Pro-inflammatory plasma cytokines were increased, but the plasma adipokine, adiponectin, was decreased in vehicle-treated septic mice. This corresponded with inhibitor κB (IκBα) protein degradation and an increase in NF-κB activity in lung. Pioglitazone treatment improved plasma Glc and adiponectin levels, and decreased pro-inflammatory cytokines. Lung IκBα protein expression increased and corresponded with a decrease in NF-κB activity in the lung from pioglitazone-treated mice. Pioglitazone reduces the inflammatory response in polymicrobial sepsis in part through inhibition of NF-κB and may be a novel therapy in sepsis.
A single short session of media perfusion induces osteogenesis in hBMSCs cultured in porous scaffolds, dependent on cell differentiation stage

Perfusing culture media through porous cell‐seeded scaffolds is now a common approach within many tissue engineering strategies. Human bone‐marrow derived mesenchymal stem cells (hBMSC) are a clinically valuable source of osteoprogenitors that respond to mechanical stimuli. However, the optimal mechanical conditions for their osteogenic stimulation in vitro have not been defined. Whereas the effects of short durations of media fluid flow have been studied in monolayers of osteoblastic cells, in 3D culture continuous or repeated perfusion is usually applied. Here, we investigated whether a short, single perfusion session applied to hBMSCs cultured in 3D would enhance their osteogenesis in vitro. We cultured hBMSCs on gelatine‐coated, porous polyurethane scaffolds with osteogenic supplements and stimulated them with a single 2‐h session of unidirectional, steady, 2.5 mL/min media perfusion, at either early or late stages of culture in 3D. Some cells were pre‐treated in monolayer with osteogenic supplements to advance cell differentiation, followed by 3D culture also with the osteogenic supplements. We report that this single, short session of media perfusion can markedly enhance the expression of bone‐related transcription and growth factors, and matrix components, by hBMSCs but that it is more effective when cells reach the pre‐osteoblast or osteoblast differentiation stage. These findings could aid in the optimization of 3D culture protocols for efficient bone tissue engineering. Biotechnol. Bioeng. 2016;113: 1814–1824. © 2016 Wiley Periodicals, Inc.
[Effects of APOC3 polymorphisms on the plasma lipids in healthy adolescents with different body mass index].

OBJECTIVE To investigate the possible effects of apolipoprotein C I gene (APOC3) polymorphisms on plasma lipids in healthy adolescents with different body mass index (BMI). METHODS Seven hundred and twenty three adolescents were divided into four groups according to BMI: group 1 CBMI= (17.80 +/- 0.75) kg/m2,n=180], group 2 [BMI = (19.39 +/- 0.32) kg/m2, n=182), group 3 [BMI= (20.68 +/- 0.43) kg/m2, n=1813 and group 4 [BMI= (23.40 +/- 2.05) kg/m2 ,n=180J. Fasting venous blood samples were collected, plasma lipids were determined and genome DNA was extracted for determining the genotypes of the APOC3 Sst I and -482C>T polymorphisms by PCR-RFLP. RESULTS With the elevation of BMI, height and plasma high-density lipoprotein cholesterol decreased significantly (P<0.001 for both), body mass, waist circumference, hip circumference, waist/hip ratio, plasma triglycerides (TG), total cholesterol and low-density lipoprotein cholesterol levels increased significantly (P<0.001 for all). No significant differences in TG levels among Sst I genotypes were observed in group 1, group 2 and group 3; but in group 4, significant differences in TG levels among Sst I genotypes were observed, S2 carriers had higher TG levels than the adolescents with S1S1 genotype. No significant differences in plasma lipids among -482C>T genotypes were observed in all groups. CONCLUSION The elevation of plasma TG levels by the S2 allele of APOC3 Sst I polymorphism is associated with BMI. It is possible that the reduction of body mass could favorably modulate the elevation of TG levels by S2 allele in healthy adolescents.