Miracle-dz/UniFAR-FTUs huggingface dataset:
Topology of NAT2, a Prototypical Example of a New Family of Amino Acid Transporters*

Amino acids are the predominant form of nitrogen available to the heterotrophic tissues of plants. These essential organic nutrients are transported across the plasma membrane of plant cells by proton-amino acid symporters. Our lab has cloned an amino acid transporter from Arabidopsis, NAT2/AAP1, that represents the first example of a new class of membrane transporters. We are investigating the structure and function of this porter because it is a member of a large gene family in plants and because its wide expression pattern suggests it plays a central role in resource allocation. In the results reported here, we investigated the topology of NAT2 by engineering a c-myc epitope on either the N or C terminus of the protein. We then used in vitro translation, partial digestion with proteinase K, and immunoprecipitation to identify a group of oriented peptide fragments. We modeled the topology of NAT2 based on the lengths of the peptide fragments that allowed us to estimate the location of protease accessible cleavage sites. We independently identified the location of the N and C termini using immunofluorescence microscopy of NAT2 expressed in COS-1 cells. We also investigated the glycosylation status of several sites of potentialN-linked glycosylation. Based on the combined data, we propose a novel 11 transmembrane domain model with the N terminus in the cytoplasm and C terminus facing outside the cell. This model of protein topology anchors our complementary investigations of porter structure and function using site-directed and random mutagenesis.
Does Loving a Brand Mean Loving Its Products? the Role of Brand-Elicited Affect in Brand Extension Evaluations

Three studies examine the influence of brand-elicited affect on consumers' evaluations of brand extensions. When a brand spontaneously elicits affective reactions, consumers appear to form an initial impression of the brand's new extension based on these reactions. The affect that they experience for other reasons and attribute to the brand can influence this impression as well. Their later evaluations of the extension are then based on this impression. This is true regardless of the similarity between the extension and the core brand. These results contrast with evidence that affect influences brand extension evaluations through its mediating impact on perceptions of core-extension similarity. This latter influence occurs only when consumers are explicitly asked to estimate the extension's similarity to the core before they evaluate it.
Genetics of the white-opaque transition in Candida albicans: demonstration of switching recessivity and mapping of switching genes

Spheroplast fusion has been used to analyze the genetics of the reversible phenotypic transition, white-opaque, in Candida albicans WO-1. This transition involves changes in cell shape, permeability, and colony morphology. Fusion of switching with nonswitching cells gave nonswitching fusants, suggesting that the white-opaque phenotype is recessive. Chromosome loss induced by heat shock gave segregants of the fusants which were able to undergo the transition, indicating that the repressor function is genetically defined and probably limited to one or two chromosomes. Chromosomes R, 1, 3, 4, and 7 were eliminated as unique sites for the repressor, leaving 2, 5, and 6 as possible locations. When a ura3 (chromosome 3) nonswitching strain was fused with a switching strain, all ura3 segregants induced by heat shock were incapable of the phenotypic transition. Therefore, some or all of the genes (called SWI genes) essential for the transition are located on chromosome 3. UV irradiation-induced recombination did give rise to Ura- switching progeny, showing that the failure to switch was not due to a side effect of the pyrimidine requirement. The failure to isolate normally switching ura3 progeny generated by UV irradiation suggests a close linkage between the two genes.
In serum‐free culture thyroid hormones can induce full expression of chondrocyte hypertrophy leading to matrix calcification

A serum‐free culture system has been developed to examine the biologic factors involved in the regulation of cellular maturation, extracellular matrix assembly, and calcification in the physis of the bovine fetal growth plate. Isolated prehypertrophic chondrocytes in high density culture undergo a process of cellular maturation whereby full expression of the hypertrophic phenotype is characterized first by type X collagen synthesis followed by matrix calcification. Using this culture system, we compared the capacity of tri‐iodothyronine (T3) with thyroxine (T4) to stimulate expression of the hypertrophic phenotype and matrix calcification in three (B, C, and D) maturationally distinct prehypertrophic chondrocyte subpopulations. The B cell subpopulation was the most mature followed by C and D subpopulations in order of decreasing maturity. Comparisons were made to cultures in fetal calf serum (FCS). In Dulbecco's modified Eagle's medium supplemented with insulin, transferrin, and selenium, both hormones (T3/T4) separately induced, in a dose‐dependent manner, chondrocyte maturation to the hypertrophic phenotype characterized by increased type X collagen mRNA and induction of protein synthesis of this molecule, together with increased alkaline phosphatase activity, and eventually calcification of the extracellular matrix. Such cellular maturation to the hypertrophic phenotype was not observed in the absence of T3 or T4 with subpopulations C and D. Only in older fetuses (>210 days) was this observed and then only in the B subpopulation. Furthermore, T3 was at least 50‐fold more potent than T4. The effects of T3 were most pronounced with the most immature cells (subpopulations C and D) where, in the case of the subpopulation C, in contrast to 0.5 nM T3 50 nM T4 was unable to induce expression of the hypertrophic phenotype. Alkaline phosphatase activity was also increased in the C cell subpopulation treated with 1 nM T3 (35.5 U/μg of DNA) over that supplemented with 50 nM T4 (7.8 U/μg of DN A). Furthermore, matrix calcification, measured by the incorporation of 45Ca2+ into the cell layer, always occurred earlier in cells cultured with T3 compared with T4. Cellular maturation to the hypertrophic phenotype was not accompanied by significant changes in DNA content; this ordinarily increases during culture in the presence of serum. Compared with cells cultured in the presence of serum, either thyroid hormone more potently induced cellular maturation. This study demonstrates that the most immature chondrocytes at the prehypertrophic stage are direct targets for T3 and T4 and, to a much a lesser degree, that either hormone is able to induce full chondrocyte hypertrophy from an early maturational stage leading to matrix calcification. But T3 is much more potent than T4. These studies also offer a new serum‐free chemically defined medium containing T3 or T4 for the culture of defined prehypertrophic chondrocytes that supports matrix assembly, hypertrophic expression, followed by matrix calcification.
Dose-response relationship of intrathecal morphine for postcesarean analgesia.

BACKGROUND This series investigated the quality of analgesia and the incidence and severity of side effects of intrathecal morphine for post-cesarean analgesia administered over a dose range of 0.0-0.5 mg. METHODS ONE hundred eight term parturients undergoing cesarean delivery at term and given spinal anesthesia were randomized to receive a single dose of intrathecal morphine (0.0, 0.025, 0.05, 0.075, 0.1, 0.2, 0.3, 0.4, or 0.5 mg). A patient-controlled analgesia (PCA) device provided free access to additional analgesics. PCA morphine use, incidence and severity of side effects, and need for treatment interventions were recorded for 24 h. Data were analyzed with analysis of variance and linear regression analysis for trends among groups. RESULTS Patient-controlled analgesia use differed significantly between groups; PCA use was higher in the control group than in groups receiving 0.075, 0.1, 0.3, 0.4, or 0.5 mg. Twenty-four-hour PCA morphine use was 45.7 mg lower (95% CI, 4.8-86.6 mg lower) in the 0.075-mg group than the control group. There was no difference in PCA morphine use between the 0.075- and 0.5-mg groups (95% CI, 36.8 mg lower to 45.0 mg higher); despite a fivefold increase in intrathecal morphine dose, PCA morphine use remained constant. There was no difference between control and treatment groups or among treatment groups with respect to nausea and vomiting. Pruritus and the need for treatment interventions increased in direct proportion to the dose of intrathecal morphine (linear regression, P = 0.001 and P = 0.0002, respectively). CONCLUSIONS These data indicate there is little justification for use of more than 0.1 mg for post-cesarean analgesia. For optimal analgesia, augmentation [corrected] of intrathecal morphine with systemic opioids may be necessary.
Arthroscopically Assisted Reconstruction of the Anterior Cruciate Ligament: A Follow-up Report

Background: The purpose of this study was to further delineate the outcome of arthroscopically assisted anterior cruciate ligament reconstruction in 125 patients who had previously been followed for two to five years. One of the original 125 patients was excluded from the present study because of insufficient follow-up, and an additional group of 101 patients was added. All 225 patients in the present study were followed for a minimum of six years. Methods: Patients were randomly assigned to reconstruction with a double-stranded semitendinosus-gracilis graft with use of a two‐incision technique (group I), reconstruction with a patellar ligament graft with use of a two‐incision technique (group II), or reconstruction with a patellar ligament graft with use of a single‐incision endoscopic technique (group III). The groups were compared with regard to the rate of graft failure, the amount of instability, knee strength, radiographic signs of degenerative changes, and functional outcome. Results: There was no significant difference among the three groups with regard to the rate of graft failure, the amount of knee instability, or the functional outcome. A normal or nearly normal functional outcome was recorded for 208 (92%) of the 225 patients. There were significant differences among the groups with regard to quadriceps muscle-strength deficits: group I had fewer patients with deficits than group III, and groups I and III both had fewer patients with deficits than group II (p = 0.04). There also were significant differences among the groups with regard to hamstring muscle-strength deficits: group III had fewer patients with deficits than group II, and group II had fewer patients with deficits than group I (p < 0.01). Twelve knees (16%) in group I, six knees (8%) in group II, and eight knees (11%) in group III showed radiographic evidence of progressive degenerative changes, but the differences among the three groups were not significant. Conclusion: Although 11.6% of the 225 knees had radiographic evidence of degenerative arthritis at a minimum of six years after arthroscopically assisted reconstruction of the anterior cruciate ligament, the choice of graft and the technique of reconstruction did not seem to affect the rate of development of these changes.
Cold-Water Immersion and Contrast Water Therapy: No Improvement of Short-Term Recovery After Resistance Training.

CONTEXT An athlete's ability to recover quickly is important when there is limited time between training and competition. As such, recovery strategies are commonly used to expedite the recovery process. PURPOSE To determine the effectiveness of both cold-water immersion (CWI) and contrast water therapy (CWT) compared with control on short-term recovery (<4 h) after a single full-body resistance-training session. METHODS Thirteen men (age 26 ± 5 y, weight 79 ± 7 kg, height 177 ± 5 cm) were assessed for perceptual (fatigue and soreness) and performance measures (maximal voluntary isometric contraction [MVC] of the knee extensors, weighted and unweighted countermovement jumps) before and immediately after the training session. Subjects then completed 1 of three 14-min recovery strategies (CWI, CWT, or passive sitting [CON]), with the perceptual and performance measures reassessed immediately, 2 h, and 4 h postrecovery. RESULTS Peak torque during MVC and jump performance were significantly decreased (P < .05) after the resistance-training session and remained depressed for at least 4 h postrecovery in all conditions. Neither CWI nor CWT had any effect on perceptual or performance measures over the 4-h recovery period. CONCLUSIONS CWI and CWT did not improve short-term (<4-h) recovery after a conventional resistance-training session.
INSM1 Demonstrates Superior Performance to the Individual and Combined Use of Synaptophysin, Chromogranin and CD56 for Diagnosing Neuroendocrine Tumors of the Thoracic Cavity

Despite the importance of recognizing neuroendocrine differentiation when diagnosing tumors of the thoracic cavity, the sensitivity of traditional neuroendocrine markers is suboptimal, particularly for high-grade neuroendocrine carcinomas such as small cell lung carcinoma and large cell neuroendocrine carcinoma. To increase sensitivity, neuroendocrine markers are routinely ordered as panels of multiple immunostains where any single positive marker is regarded as sufficient evidence of neuroendocrine differentiation. Insulinoma-associated protein 1 (INSM1) is a well-validated transcription factor of neuroendocrine differentiation that has only recently been evaluated for diagnostic use. We performed INSM1 immunohistochemistry on a large series of thoracic neuroendocrine and non-neuroendocrine tumors and compared its performance to synaptophysin, chromogranin, and CD56. INSM1 was positive in 94.9% of small cell lung carcinomas and 91.3% of large cell neuroendocrine carcinomas, compared with 74.4% and 78.3% with the combined panel of traditional markers. INSM1 also stained all (100%) of the atypical carcinoids, typical carcinoids and mediastinal paragangliomas, but only 3.3% of adenocarcinomas and 4.2% of squamous cell carcinomas. Overall, INSM1 demonstrated a sensitivity of 96.4% across all grades of thoracic neuroendocrine tumors, significantly more than the 87.4% using the panel of traditional markers (P=0.02). INSM1 is sufficiently sensitive and specific to serve as a standalone first-line marker of neuroendocrine differentiation. A more restrained approach to immunohistochemical analysis of small thoracic biopsies is appropriate given the expanding demand on this limited material for therapeutic biomarker analysis.
Risk factors for intramammary infections and subclinical mastitis in post-partum dairy heifers

The prevalence of intramammary infections (IMI) and subclinical mastitis (SCM) in 436 German Holstein heifers was put in relation with clinical findings of the udder and data regarding individual rearing and housing conditions of the animals. The clinical examination took place on the day of the livestock auction (at approximately 41 d in milk, DIM). On that day, 31% of the heifers had IMI in at least one quarter, and 18% of all quarters were infected. Coagulase-negative staphylococci were the most prevalent bacteria isolated, accounting for 68% of the positive samples. Data were analysed by logistic regression. Criteria such as ‘juvenile intersucking’, ‘teats shorter than 35 mm’, ‘teats with a diameter <18 mm’ and ‘udder oedema at the day of the auction’ were associated with IMI in heifers during the first 41 DIM. Loose-housing systems during pregnancy (as opposed to tie-stalls), juvenile intersucking, clinical mastitis during the first week after calving, teat diameters <18 mm, and employing organic bedding material in the stables before calving were associated with subclinical mastitis.
Elevated adrenomedullin mRNA in lymphoblastoid cells from schizophrenic patients

Adrenomedullin (ADM) is a 52 amino acid peptide with multiple physiological functions and wide tissue distributions including brain. Recently, elevated plasma levels of ADM were found in patients with schizophrenia, bipolar affective disorder and autism, suggesting the involvement of ADM in the pathophysiology of mental diseases. Using real-time quantitative PCR, we compared the ADM mRNA levels in lymphoblastoid cell lines between schizophrenic patients and controls. Male but not female schizophrenia patients had 2- to 3-fold higher ADM mRNA levels than controls (p<0.01). Our data support that ADM may be associated with the pathophysiology of schizophrenia, although the cause of the association needs further study.
The Control of Politicians in Normal Times and Times of Crisis: Wealth Accumulation by U.S. Congressmen, 1850-1880

We employ a regression discontinuity design based on close elections to estimate the rents from a seat in the U.S. congress between 1850-1880. Using census data, we compare wealth accumulation among those who won or lost their first race by a small margin. We find evidence of significant returns for the first half of the 1860s, during the Civil War, but not for other periods. We hypothesize that increased opportunities from the sudden spike in government spending during the war and the decrease in control by the media and other monitors might have made it easier for incumbent congressmen to collect rents.
Transplantation of neural stem cells overexpressing glial cell line‐derived neurotrophic factor enhances Akt and Erk1/2 signaling and neurogenesis in rats after stroke

Background Our previous studies have indicated that the beneficial effects of grafting neural stem cells (NSCs) overexpressing glial cell line‐derived neurotrophic factor (GDNF) in rats after stroke. However, the underlying mechanisms are highly debatable. In this study, we investigated whether neurogenesis, Akt, and extracellular signal‐regulated kinase 1/2 (Erk1/2) signaling were involved in this process. Methods Transient ischemic stroke were induced by occluding middle cerebral artery for 2 hours and reperfusion. At 3 days after reperfusion, GDNF/NSCs, NSCs, and vehicle were administered. Immunohistochemical staining was used to evaluate neurogenesis by nestin antibody; phosphorylation of Akt and Erk1/2 was investigated by Western blotting analysis. Results Transplantation of GDNF/NSCs and NSCs significantly increased nestin‐positive cells compared to control group (vehicle) from 1 to 7 weeks after reperfusion, and GDNF/NSCs showed stronger effect than NSCs at 2 and 3 weeks after reperfusion. Meanwhile, enhanced phosphorylation level of Erk1/2 was observed in the GDNF/NSCs and NSCs groups compared with control group, and phosphorylation level of Erk1/2 in GDNF/NSCs group was remarkably higher than that of NSCs group at any given time. In contrast, expression of mitogen‐activated protein kinase phosphatase‐1 (MKP‐1), known as inhibitor of Erk1/2 signaling, was significantly decreased in the GDNF/NSCs and NSCs groups compared with the control group. Moreover, much enhanced and prolonged phosphorylation level of Akt of GDNF/NSCs group was detected compared with control and NSCs group. Conclusion Grafting GDNF/NSCs enhances neurogenesis and activates Akt and Erk1/2 signaling, that may provide the potential for GDNF/NSCs in stroke treatment.
Dysphagia caused by Anterior Cervical Osteophyte: A Rare Entity Revisited

Large anterior cervical osteophytes can occur in degeneration of the cervical spine, cervical spondylosis or in diffuse idiopathic skeletal hyperostosis (DISH). Voluminous anterior cervical osteophytes which can develop from C3 to C7 can cause narrowing of the pharyngoesophageal segment by external compression and may cause dysphagia, which may be life threatening when it is associated with aspiration and or dyspnea. The objective of this case report is to highlight how commonly occurring anterior cervical osteophytes may become an unrecognized cause for life-threatening dysphagia. The clinical and radiographic findings in patient with dysphagia and ventral osteophytes of the cervical spine due to degeneration are demonstrated. The anterolateral approach for removal of these osteophytes is described.
EFFECTS OF RECIPROCAL PEER-QUESTIONING INSTRUCTION ON EFL COLLEGE STUDENTS ’ ENGLISH

This study investigated the effects of reciprocal peer-questioning instruction to enhance the English reading comprehension of English as a foreign language (EFL) college students by comparison with the conventional lecture instruction. We employed a pretest-posttest quasi-experimental comparison group design in college English reading courses in a complete semester. A total of 78 EFL college students enrolled in compulsory English reading courses participated in this study; 38 participants were in the experimental group and 40 were in the comparison group. The experimental group received reciprocal peer-questioning instruction, whereas the comparison group received conventional lecture instruction. The data were analyzed by mean, standard deviation, t tests, and one-way ANCOVA. The findings indicate statistically significant differences in favor of reciprocal peer-questioning instruction on English reading comprehension, particularly among highand medium-proficiency students. Compared with conventional lecture instruction, reciprocal peer-questioning instruction created a more positive attitude toward learning English reading. In conclusion, we strongly recommend EFL instructors use reciprocal peer-questioning instruction in college English reading classrooms. © 2014 AESS Publications. All Rights Reserved.
Oestrogen receptor α genotype, and interactions between vitamin D receptor and transforming growth factor‐β1 genotypes are associated with quantitative calcaneal ultrasound in postmenopausal women

objective Quantitative ultrasound (QUS) of bone is a new radiation‐free, low‐cost method that measures both bone mass and quality. We investigated associations between QUS parameters and polymorphisms of vitamin D receptor (VDR), oestrogen receptor α (ERα) and transforming growth factor‐β1 (TGF‐β1) genes in postmenopausal women residing in a community.
Universal In Vitro Antifungal Resistance of Genetic Clades of the Fusarium solani Species Complex

ABSTRACT Eleven antifungal drugs were tested against representative isolates of the four phylogenetic clades of the Fusarium solani species complex obtained in a multilocus sequence analysis. They all showed very poor activity, with no differences among the clades. Amphotericin B was the most active drug.
Sensitivity of different evapotranspiration calculation methods in different crop-weather models

The calculation of evapotranspiration and its components transpiration and evapotranspiration is of crucial importance in dynamic crop-weather models, irrigation models and SVAT models. Although many approaches were developed and adapted for various applications and based on available input data, there is still a remarkable range of uncertainty related for a representative calculation. However, many of the problems are based on weak quality of model input data, but others are related to the method itself and its sensitivity to various environmental conditions. In our study, several methods of evapotranspiration estimation in different crop weather models are compared and tested against data from lysimeters at a location in Austria. An analysis shows differences in the results, related sensitivities in the used methods and crucial parameters for their parameterisation.