Miracle-dz/UniFAR-FTUs huggingface dataset:
Purification and Characterization of an Active N-Acetylglucosaminyltransferase Enzyme Complex from Streptococci

ABSTRACT A new family of bacterial serine-rich repeat glycoproteins can function as adhesins required for biofilm formation and pathogenesis in streptococci and staphylococci. Biogenesis of these proteins depends on a gene cluster coding for glycosyltransferases and accessory secretion proteins. Previous studies show that Fap1, a member of this family from Streptococcus parasanguinis, can be glycosylated by a protein glycosylation complex in a recombinant heterogeneous host. Here we report a tandem affinity purification (TAP) approach used to isolate and study protein complexes from native streptococci. This method demonstrated that a putative glycosyltransferase (Gtf2), which is essential for Fap1 glycosylation, readily copurified with another glycosyltransferase (Gtf1) from native S. parasanguinis. This result and the similar isolation of a homologous two-protein complex from Streptococcus pneumoniae indicate the biological relevance of the complexes to the glycosylation in streptococci. Furthermore, novel N-acetylglucosaminyltransferase activity was discovered for the complexes. Optimal activity required heterodimer formation and appears to represent a novel type of glycosylation.
Heritability of Anthropometric Phenotypes in Caste Populations of Visakhapatnam, India

In this study, we used anthropometric data from six Andhra caste populations to examine heritability patterns of 23 anthropometric phenotypes (linear, craniofacial, and soft tissue measures) with special reference to caste differences. We obtained anthropometric data from 342 nuclear families from Brahmin, Reddy, Telaga, Nagara, Ag. Kshatriya, and Mala castes of Visakhapatnam, India. These caste groups represent the existing hierarchical stratification of Indian populations. We used a variance components approach to determine the heritability (h2) of these 23 anthropometric phenotypes (height, weight, BMI, etc.). The sample consisted of 1918 individuals ranging in age from 6 to 72 years (mean = 21.5, S.D. = 13.8). The heritabilities (h2 ± S.E.) for all anthropometric traits for the entire sample were significant (p < 0.0001) and varied from 0.25 ± 0.05 (BMI) to 0.61 ± 0.05 (bizygomatic breadth) after accounting for sex, age, and caste effects. Since data on socioeconomic and nutritional covariates were available for a subset of families, we repeated the genetic analyses using this subset, which has yielded higher heritabilities ranging from 0.21 ± 0.16 (head breadth) to 0.72 ± 0.18 (nasal breadth). In general, craniofacial measurements exhibited higher h2 compared to linear measures. Breadth measurements and circumferences yielded more or less similar heritabilities. Age and sex effects were significant (p < 0.0001) for most of the traits, while the effects of caste, socioeconomic status, and nutritional status were inconsistent across the traits. In conclusion, anthropometric phenotypes examined in this study are under appreciable additive genetic influences.
A microfluidic device that forms and redirects pheromone gradients to study chemotropism in yeast.

Chemotropism, or directed cell growth in response to a chemical gradient, is integral to many biological processes. The mating response of the budding yeast, Saccharomyces cerevisiae, is a well studied model chemotropic system. Yeast cells of opposite mating type signal their positions by secreting soluble mating pheromones. The mutual exchange of pheromones induces the cells to grow towards one another, resulting in mating projections or "shmoos. " Yeast cells exhibit a remarkable ability to orient their growth toward the nearest potential mating partner, and to reorient (i.e., bend their mating projections) in response to a change in the direction of the pheromone gradient. Although a number of microfluidic devices have been used to generate linear pheromone gradients and to measure initial orientation, none of them have the capability to change the direction of the gradient, other than to invert it. We have developed a microfluidic device that can produce stable pheromone gradients and rapidly rotate them in 90° increments, mimicking the dynamic gradients yeast are exposed to in situ, and allowing for the study of reorientation as well as initial orientation. The mean angle of orientation exhibited by gradient-stimulated yeast cells in this device was 56.9°. In control experiments, cells subjected to pheromone coming from all four directions showed no evidence of orientation. Switching the direction of the pheromone source by 90° induced 83.6% of the polarized cells to change their direction of growth. Of these, 85.2% bent their mating projections toward the second source, demonstrating the utility of this device in the study of reorientation with specifically controlled gradients.
Glycerophosphoinositol, a Novel Phosphate Source Whose Transport Is Regulated by Multiple Factors in Saccharomyces cerevisiae*

Git1p mediates the transport of the phospholipid metabolite, glycerophosphoinositol, into Saccharomyces cerevisiae. We report that phosphate limitation and inositol limitation affect GIT1 expression and Git1p transport activity via distinct mechanisms that involve multiple transcription factors. GIT1 transcript levels and Git1p activity are greater in cells starved for phosphate, with or without inositol limitation, than in cells only limited for inositol. Furthermore, the kinetics of GIT1 transcript accumulation and Git1p activity upon transfer of cells to phosphate starvation media are different from those obtained upon transfer of cells to inositol-free media. Pho2p and Pho4p are required for GIT1 expression and for Git1p transport activity under all growth conditions tested. In contrast, Ino2p and Ino4p are required for full GIT1 expression when inositol is limiting, with or without phosphate limitation, but not when only phosphate is limiting. Greatly reduced transport activity was detected in ino2Δ and ino4Δ cells under all growth conditions. A 300-base pair region of the GIT1 promoter containing potential Pho4p binding sites was shown to be required for full GIT1 expression. Git1p appears to act as a H+-symporter, and neither inositol nor phosphate effectively compete with glycerophosphoinositol for transport by Git1p. Glycerophosphoinositol was shown previously to support the growth of an inositol auxotroph. Remarkably, we now report that glycerophosphoinositol can act as the sole source of phosphate for the cell, providing functional relevance for the regulation of Git1p transport activity by phosphate.
Metabolism of [14C]benzyl selenocyanate in the F344 rat.

Benzyl selenocyanate (BSC), a synthetic organoselenium compound, has been shown to inhibit chemically induced tumors in several animal model systems. However, it is not known whether BSC or one of its metabolites is responsible for the chemopreventive effect. An initial approach to this question requires the structural elucidation of BSC metabolites in vitro and in vivo. To determine the structures of BSC metabolites in vitro, we studied the metabolism of [14C]BSC using Aroclor-induced rat liver 9000g supernatant. Under these conditions, BSC was partially converted to dibenzyl diselenide (DDS) and phenylmethaneseleninic acid. The metabolism of [14C]BSC (12.5 mg/kg body weight, 8 mCi/mmol, oral administration) in male F344 rats was also studied. Excretion was monitored by measurement of radioactivity as well as by selenium content using atomic absorption spectrophotometry (AAS). The results indicate that urine was the major route of excretion. Approximately 22% of the dose was excreted in the urine over the course of 35 days; however, a large portion (approximately 70%) of the dose remained in the body. Benzoic acid, hippuric acid, and their sulfate and glucuronide conjugates, accounting for 16% of the dose, were identified in the urine. The formation of these metabolites indicates that BSC is metabolized in part via bond cleavage between the benzyl moiety and the selenocyanate function. Additional support for this cleavage was obtained from fecal analysis; over the course of 23 days 9% of the selenium (AAS) but only less than 1% of the radioactivity was recovered in feces. No radioactivity was detected in the exhaled air. We also studied the metabolism of [14C]DDS (17.3 mg/kg body weight, 2.5 mCi/mmol) in male F344 rats. (ABSTRACT TRUNCATED AT 250 WORDS)
EFFECTS OF NUMERACY AND FINANCIAL LITERACY ON UNDESIRABLE FINANCIAL AND RELATED ACADEMIC OUTCOMES IN COLLEGE STUDENTS

EFFECTS OF NUMERACY AND FINANCIAL LITERACY ON UNDESIRABLE FINANCIAL AND RELATED ACADEMIC OUTCOMES IN COLLEGE STUDENTS Sriram Govindarajan, Ph. D. Supervising Professor: Daniel S. Levine Numeracy was a better predictor of financial delinquency than financial literacy. The relationship between numeracy and undesirable academic effects was fully mediated by financial delinquency. It was also found that in a credit card related decision making task, the relationship between numeracy and performance was partially mediated by the deliberation time. This is similar to the finding in Ghazal et al. (2014) and supporting their conclusion that numeracy is just not facility with numbers, but also related directly to heuristic deliberation and metacognition. The findings also support the notion that people low in numeracy might be prone to various biases and fallacies that might lead to deleterious financial behavior and attitudes. NUMERACY AND FINANCIAL OR ACADEMIC OUTCOMES 5
Draft Genome Sequences of Vibrio sp. Strains Isolated from Tetrodotoxin-Bearing Scavenging Gastropod

ABSTRACT Vibrio sp. strains JCM 18905 and JCM 19053 were isolated from a tetrodotoxin (TTX)-bearing scavenging gastropod, and Vibrio sp. strain JCM 18904 was isolated from a sea cucumber. All these are closely related to Vibrio alginolyticus. Their comparative genome information is useful for studies of TTX production in bacteria.
New method for generating deletions and gene replacements in Escherichia coli

We describe a method for generating gene replacements and deletions in Escherichia coli. The technique is simple and rapid and can be applied to most genes, even those that are essential. What makes this method unique and particularly effective is the use of a temperature-sensitive pSC101 replicon to facilitate the gene replacement. The method proceeds by homologous recombination between a gene on the chromosome and homologous sequences carried on a plasmid temperature sensitive for DNA replication. Thus, after transformation of the plasmid into an appropriate host, it is possible to select for integration of the plasmid into the chromosome at 44 degrees C. Subsequent growth of these cointegrates at 30 degrees C leads to a second recombination event, resulting in their resolution. Depending on where the second recombination event takes place, the chromosome will either have undergone a gene replacement or retain the original copy of the gene. The procedure can also be used to effect the transfer of an allele from a plasmid to the chromosome or to rescue a chromosomal allele onto a plasmid. Since the resolved plasmid can be maintained by selection, this technique can be used to generate deletions of essential genes.
Perceptions of middle, high school and community college agricultural teachers in Iowa regarding agricultural awareness concepts and activities in the middle school curriculum: Grades 7-9

Teachers are relied upon to teach basic agricultural knowledge among young learners who are devoid of real-world agricultural experiences. The middle school serves as a tool for achieving this goal because self and environmental awareness, and career choices begin at this educational stage. Since agricultural teachers are the major driver of agricultural education program in the middle school, their perceptions regarding this program are important. The purpose of this study was to determine the perceptions of agricultural teachers regarding teaching agriculture and selected course content in the middle school. The Theory of Planned Behavior was used as a theoretical framework to predict teachers’ perceptions regarding teaching agriculture in the middle school. Questionnaires were sent to middle school, high school and community college agriculture teachers in Iowa through Qualtrics. The results of the study showed that agriculture teachers in Iowa had positive and strong perceptions toward teaching agriculture in the middle school. In addition, agricultural teachers in Iowa indicated selected content of the Agriculture, Food, and Natural Resources curriculum developed by the National Association of Agricultural Education should be taught to a limited or great extent. Similarly, agricultural teachers in Iowa believed teaching agricultural education in the middle school will develop students’ leadership skills and assist students in making informed career choices in agricultural industry. Based on the findings of this study, the cyclical model was developed as a guide for curriculum development, and planning of instructional activities. Teaching agriculture in the middle school develops, trains, and motivates young students to become responsible citizens and capable future policy makers. Participation in curricular and non-curricular agricultural activities could promote the development of student leadership skills. The positive perceptions of agriculture teachers regarding teaching agriculture in the middle school indicate its sustainability and positive impacts on students.
Impaired long‐term memory and long‐term potentiation in N‐type Ca2+ channel‐deficient mice

Voltage‐dependent N‐type Ca2+ channels, along with the P/Q‐type, have a crucial role in controlling the release of neurotransmitters or neuromodulators at presynaptic terminals. However, their role in hippocampus‐dependent learning and memory has never been examined. Here, we investigated hippocampus‐dependent learning and memory and synaptic plasticity at hippocampal CA3–CA1 synapses in mice deficient for the α1B subunit of N‐type Ca2+ channels. The mutant mice exhibited impaired learning and memory in the Morris water maze and the social transmission of food preference tasks. In particular, long‐term memory was impaired in the mutant mice. Interestingly, among activity‐dependent long‐lasting synaptic changes, theta burst‐ or 200‐Hz‐stimulation‐induced long‐term potentiation (LTP) was decreased in the mutant, compared with the wild‐type mice. This type of LTP is known to require brain‐derived neurotrophic factor (BDNF). It was found that both BDNF‐induced potentiation of field excitatory postsynaptic potentials and facilitation of the frequency of miniature excitatory postsynaptic currents (mEPSCs) were reduced in the mutant. Taken together, these results demonstrate that N‐type Ca2+ channels are required for hippocampus‐dependent learning and memory, and certain forms of LTP.
Political determinants of social expenditures in Greece: an empirical analysis

A view prominently expounded is that the interaction between the composition and the volume of public expenditures is directly affected by political, institutional, psephological and ideological indicators. A crucial component of public expenditures, social expenditures play an important role in the economy as they directly and indirectly affect the distribution of income and wealth. Social expenditures aim at reallocating income and wealth unequal distribution. These expenditures comprise cash benefits, direct in-kind provision of goods and services, and tax breaks with social purposes.The aim of this study is to determine the relationship between political structure, i.e. government fragmentation, ideological composition, elections and so on, and the social expenditures in Greece. Employing data from the Comparative Political Dataset (CPDS) and the OECD Social Expenditure Database (SOCX), a time series analysis was conducted for Greece for the 1980-2014 period. The findings of the study indicate that voter turnout, spending on the elderly population and the number of government changes have positive and statistically significant effects on social expenditures in Greece while debt stock and cabinet composition have negative effects.
Quantification of myocardial blood flow using 13N-ammonia and PET: comparison of tracer models.

UNLABELLED Several tracer kinetic methods have been proposed for quantification of regional myocardial blood flow (MBF) with 13N-ammonia and PET. Merits and limitations specific to each approach, however, generally are not clear, because they have not been evaluated in the same experimental environment. Therefore, we compared six different commonly used methods (11 modifications) to characterize the accuracy of each approach. The methods included the two-parameter model (method 1), the modified two-parameter model (method 2), the four-parameter model (method 3), the graphical analysis (method 4), the first-pass extraction method (method 5) and the dose uptake index (DUI; method 6). METHODS Eleven studies in four dogs, 16 studies in eight healthy human volunteers and 14 studies in seven patients were performed using 13N-ammonia and PET. MBF in dogs was varied with dipyridamole and coronary occlusions and was measured independently and simultaneously with microspheres. Volunteers and patients were studied at baseline and after dipyridamole. MBF and DUI were estimated using a time-activity curve (Qi[t]) derived from dynamic images and regions of interest (ROls) and using the six methods. DUI was defined as Qi(t = 2 min) x weight/dose. RESULTS MBF estimated by methods 1-5 correlated well with microsphere MBF in dogs. MBF estimates by method 1 correlated well with those by methods 2, 4 and 5 and to a lesser degree with those by method 3 in both dog and human studies. DUI correlated poorly with MBF by microspheres and by methods 1-5 in both dog and human studies. MBF estimates by method 3 showed larger dispersion (SD/mean flow) and higher sensitivity to metabolites correction in arterial blood than those by methods 1, 2, 4 and 5. CONCLUSION MBF can be measured accurately using 13N-ammonia PET and tracer kinetic methods. DUI is a poor indicator of MBF values. The results indicate that preference should be given to the two-parameter model, incorporating geometrical ROI representation (method 2) among the compartment models, and to the graphical analysis (method 4) among the noncompartmental approaches.
Angiotensin‐(1–7) increases neuronal potassium current via a nitric oxide‐dependent mechanism

Angiotensin 1–7 [Ang‐(1–7)] signaling in the brain may play an important protective role in cardiovascular diseases by decreasing sympathetic output. It is generally well‐accepted that nitric oxide (NO•) in the brain functions as a sympatho‐inhibitory molecule. We hypothesized that Ang‐(1–7) increases NO• in neurons and modulates neuronal activity. We tested this hypothesis in catecholaminergic CATH.a neurons. Protein analysis revealed these neurons express the Ang‐(1–7) receptor (MasR) and three isoforms of NO• synthase (nNOS, eNOS, iNOS). Expression of MasR, nNOS, and eNOS protein markedly increased in CATH.a neurons following six days of differentiation with N6,2′‐O‐Dibutyryladenosine 3′,5′‐cyclic monophosphate sodium salt (dbcAMP, 1 mM). As measured by DAF‐FM fluorescence and confocal microscopy, Ang‐(1–7) (100 nM) increased NO• levels in differentiated CATH.a neurons (31 ± 3%; P<0.05 vs. baseline) and this response was attenuated (P<0.05) by the MasR antagonist A779 (5 ± 3%), non‐specific NOS inhibitor (L‐NAME; −5 ± 1%), and nNOS inhibitor (SMTC; −5 ± 7%). Using the whole‐cell patch‐clamp technique, we observed that Ang‐(1–7) increased outward K+ current (IKv) in differentiated CATH.a neurons (53 ± 6% increase; P<0.05 vs. vehicle). These findings indicate that Ang‐(1–7) increases nNOS‐dependent NO• levels via MasR activation and elevates IKv in differentiated CATH.a neurons. NIH P01HL062222
Evidence that the middle T antigen of polyomavirus interacts with the membrane skeleton

The transforming protein of polyomavirus, middle T antigen, is associated with cellular membranes. We have examined the subcellular location of the middle T antigen in two different cell types by fractionation and detergent phase partitioning. Middle T antigen expressed in human cells by a recombinant adenovirus was detected primarily in the membrane skeleton. Sucrose gradient fractionation revealed that the middle T antigen was associated with complexes with molecular weights of 500,000 to 1,000,000. Several markers for cytoskeleton cofractionate with these complexes, including actin, tubulin, and vimentin. Electron micrographs of membrane skeleton prepared from cells expressing middle T antigen demonstrated that this material contained primarily fibrous structures and was clearly devoid of bilayer membranes. These structures were distinct from the filamentous structures observed in fractions enriched for cytoskeleton. Consistent with a role for membrane skeleton localization in transformation, middle T antigen was detected exclusively in fractions enriched for membrane skeleton in middle T antigen-transformed Rat-2 cells. Our results may resolve the apparent difference between middle T antigen localization as determined by immunomicroscopy and that determined by subcellular fractionation.
Combat-Exposed War Veterans at Risk for Suicide Show Hyperactivation of Prefrontal Cortex and Anterior Cingulate During Error Processing

Objective Suicide is a significant public health problem. Suicidal ideation (SI) increases the risk for completed suicide. However, the brain basis of SI is unknown. The objective of this study was to examine the neural correlates of self-monitoring in individuals at risk for suicide. We hypothesized that combat veterans with a history of SI relative to those without such a history would show altered activation in the anterior cingulate cortex and related circuitry during self-monitoring. Methods Two groups of combat-exposed war veterans (13 men with and 13 men without history of SI) were studied. Both the SI and non-SI participants had two or more of the following: a) current major depressive disorder, b) current posttraumatic stress disorder, and c) history of mild traumatic brain injury, and each subject performed a validated stop task during functional magnetic resonance imaging. Error-related activation was compared between the SI and non-SI groups. Results The SI group demonstrated more error-related activation of the anterior cingulate (8256 mm3, t = 2.51) and prefrontal cortex (i.e., clusters >2048 mm3, voxelwise p < .05). The SI and non-SI participants showed similar behavioral task performance (i.e., mean error rate, F values < 0.63, p values > .43; and mean reaction times, F = 0.27, p = .61). Conclusions These findings suggest neural correlates of altered self-monitoring in individuals with a history of SI and may further suggest that functional magnetic resonance imaging could be used to identify individuals at risk for suicide before they engage in suicidal behavior. Abbreviations SI = suicidal ideation ACC = anterior cingulate cortex OEF = Operation Enduring Freedom OIF = Operation Iraqi Freedom fMRI = functional magnetic resonance imaging TBI = traumatic brain injury MDD = major depressive disorder PTSD = posttraumatic stress disorder AFNI = Analysis of Functional Neuroimages PFC = prefrontal cortex