Search

BACKGROUND: We identified a model system that exploits the inherent taste variation in early feedings to investigate food preference development. OBJECTIVE: The objective was to determine whether exposure to differing concentrations of taste compounds in milk and formulas modifies acceptance of exemplars of the 5 basic taste qualities in a familiar food matrix. Specifically, we examined the effects of consuming hydrolyzed casein formulas (HCFs), which have pronounced bitter, sour, and savory tastes compared with breast milk (BM) and bovine milk-based formulas (MFs), in which these taste qualities are weaker. DESIGN: Subgroups of BM-, MF- and HCF-fed infants, some of whom were fed table foods, were studied on 6 occasions to measure acceptance of sweet, salty, bitter, savory, sour, and plain cereals. RESULTS: In infants not yet eating table foods, the HCF group ate significantly more savory-, bitter-, and sour-tasting and plain cereals than did the BM or MF groups. HCF infants displayed fewer facial expressions of distaste while eating the bitter and savory cereals, and they and BM infants were more likely to smile while they were eating the savory cereal. In formula-fed infants eating table foods, preferences for the basic tastes reflected the types of foods they were being fed. In general, those infants who ate more food displayed fewer faces of distaste. CONCLUSIONS: The type of formula fed to infants has an effect on their response to taste compounds in cereal before solid food introduction. This model system of research investigation sheds light on sources of individual differences in taste and perhaps cultural food preferences. Copyright © 2009 American Society for Nutrition [ABSTRACT FROM AUTHOR]

Braided rivers play a significant role in replenishing groundwater, but our understanding of how these recharge rates fluctuate over time remains limited. Traditional techniques for gauging groundwater recharge are ineffective for studying complex braided river systems due to their insufficient spatiotemporal resolution. To address this gap, active-distributed temperature sensing (A-DTS) was used. This method combines fiber optic temperature measurements with an active heat source, enabling quantification of groundwater fluxes. In this study, twelve consecutive A-DTS surveys were conducted on a 100 m long hybrid fiber optic cable to a depth of 5 m beneath the Waikirikiri Selwyn River. This experiment was conducted during a period of relatively stable river stage and flow, highlighting the effectiveness of using A-DTS to measure temporal changes in groundwater recharge., (© 2023 The Author(s).)

Contamination of potable groundwater by pathogenic viruses from on-site wastewater treatment systems (OWTS) poses a serious health risk. This study investigated the attenuation and transport of rotavirus, bacteriophage MS2 and DNA-labelled-glycoprotein-coated silica nanoparticles (DGSnp) in 2 intact cores of silt loam over gravels dosed with wastewater from an OWTS at 3.53 L/day. To simulate a worst-case scenario, experiments were conducted under saturated conditions. The results from 6 experiments demonstrated that the rotavirus and DGSnp reductions were very similar and markedly greater than the MS2 reduction. This was reflected in the peak concentrations, relative mass recoveries, and temporal and spatial reduction rates. For a given log 10 reduction, the estimated soil depth required for MS2 was over twice that required for rotavirus and DGSnp. This is the first study in which DGSnp was used as a rotavirus surrogate in soil under wastewater applications. Consistent with previous studies, DGSnp showed promise at mimicking rotavirus attenuation and transport in porous media. The results suggest DGSnp could be used to assess the attenuation capacity of subsurface media to rotavirus. However, DGSnp is not conservative and will underestimate the setback distances required for rotavirus reductions by 3%. On the other hand, separation distances determined using the rotavirus parameters and criteria but based on MS2 attenuation, can be too conservative in some subsurface media. To determine safe and realistic separation distances, it would be beneficial and complementary to apply both conservative virus surrogate using MS2 bacteriophage and representative but non-conservative new virus surrogates using biomolecule-modified silica nanoparticles., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Histidine kinases (HKs) funnel diverse environmental stimuli into a single autophosphorylation event at a conserved histidine residue. The HK EnvZ is a global sensor of osmolality and cellular acid pH. In previous studies, we discovered that osmosensing in EnvZ was mediated through osmolyte-induced stabilization of the partially disordered helical backbone spanning the conserved histidine autophosphorylation site (His 243 ). Here, we describe how backbone stabilization leads to changes in the microenvironment of His 243 , resulting in enhanced autophosphorylation through relief of inhibition and repositioning of critical side chains and imidazole rotamerization. The conserved His-Asp/Glu dyad within the partially structured helix is equally geared to respond to acid pH, an alternative environmental stimulus in bacteria. This high-resolution "double-clamp" switch model proposes that a His-Asp/Glu dyad functions as an integrative node for regulating autophosphorylation in HKs. Because the His-Asp/Glu dyad is highly conserved in HKs, this study provides a universal model for describing HK function., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Unlike eukaryotes, bacteria undergo large changes in osmolality and cytoplasmic pH. It has been described that during acid stress, bacteria internal pH promptly acidifies, followed by recovery. Here, using pH imaging in single living cells, we show that following acid stress, bacteria maintain an acidic cytoplasm and the osmotic stress transcription factor OmpR is required for acidification. The activation of this response is non-canonical, involving a regulatory mechanism requiring the OmpR cognate kinase EnvZ, but not OmpR phosphorylation. Single cell analysis further identifies an intracellular pH threshold ~6.5. Acid stress reduces the internal pH below this threshold, increasing OmpR dimerization and DNA binding. During osmotic stress, the internal pH is above the threshold, triggering distinct OmpR-related pathways. Preventing intracellular acidification of Salmonella renders it avirulent, suggesting that acid stress pathways represent a potential therapeutic target. These results further emphasize the advantages of single cell analysis over studies of population averages.

β-Barrelmembrane proteins (βMPs) form barrel-shaped pores in the outer membrane of Gram-negative bacteria, mitochondria, and chloroplasts. Because of the robustness of their barrel structures, βMPs have great potential as nanosensors for single-molecule detection. However, natural βMPs currently employed have inflexible biophysical properties and are limited in their pore geometry, hindering their applications in sensing molecules of different sizes and properties. Computational engineering has the promise to generate βMPs with desired properties. Here we report a method for engineering novel βMPs based on the discovery of sequence motifs that predominantly interact with the cell membrane and appear in more than 75% of transmembrane strands. By replacing β1-β6 strands of the protein OmpF that lack these motifs with β1-β6 strands of OmpG enriched with these motifs and computational verification of increased stability of its transmembrane section, we engineered a novel βMP called OmpGF. OmpGF is predicted to form a monomer with a stable transmembrane region. Experimental validations showed that OmpGF could refold in vitro with a predominant β-sheet structure, as confirmed by circular dichroism. Evidence of OmpGF membrane insertion was provided by intrinsic tryptophan fluorescence spectroscopy, and its pore-forming property was determined by a dye-leakage assay. Furthermore, single-channel conductance measurements confirmed that OmpGF function as a monomer and exhibits increased conductance than OmpG and OmpF. These results demonstrated that a novel and functional βMP can be successfully engineered through strand replacement based on sequence motif analysis and stability calculation., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Membrane-anchored receptors are essential cellular signaling elements for stimulus sensing, propagation, and transmission inside cells. However, the contributions of lipid interactions to the function and dynamics of embedded receptor kinases have not been described in detail. In this study, we used amide hydrogen/deuterium exchange mass spectrometry, a sensitive biophysical approach, to probe the dynamics of a membrane-embedded receptor kinase, EnvZ, together with functional assays to describe the role of lipids in receptor kinase function. Our results reveal that lipids play an important role in regulating receptor function through interactions with transmembrane segments, as well as through peripheral interactions with nonembedded domains. Specifically, the lipid membrane allosterically modulates the activity of the embedded kinase by altering the dynamics of a glycine-rich motif that is critical for phosphotransfer from ATP. This allostery in EnvZ is independent of membrane composition and involves direct interactions with transmembrane and periplasmic segments, as well as peripheral interactions with nonembedded domains of the protein. In the absence of the membrane-spanning regions, lipid allostery is propagated entirely through peripheral interactions. Whereas lipid allostery impacts the phosphotransferase function of the kinase, extracellular stimulus recognition is mediated via a four-helix bundle subdomain located in the cytoplasm, which functions as the osmosensing core through osmolality-dependent helical stabilization. Our findings emphasize the functional modularity in a membrane-embedded kinase, separated into membrane association, phosphotransferase function, and stimulus recognition. These components are integrated through long-range communication relays, with lipids playing an essential role in regulation., (Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.)

Sea water intrusion into aquifers is problematic in many coastal areas. The physics and chemistry of this issue are complex, and sea water intrusion remains challenging to quantify. Simple assessment tools like analytical models offer advantages of rapid application, but their applicability to field situations is unclear. This study examines the reliability of a popular sharp-interface analytical approach for estimating the extent of sea water in a homogeneous coastal aquifer subjected to pumping and regional flow effects and under steady-state conditions. The analytical model is tested against observations from Canada, the United States, and Australia to assess its utility as an initial approximation of sea water extent for the purposes of rapid groundwater management decision making. The occurrence of sea water intrusion resulting in increased salinity at pumping wells was correctly predicted in approximately 60% of cases. Application of a correction to account for dispersion did not markedly improve the results. Failure of the analytical model to provide correct predictions can be attributed to mismatches between its simplifying assumptions and more complex field settings. The best results occurred where the toe of the salt water wedge is expected to be the closest to the coast under predevelopment conditions. Predictions were the poorest for aquifers where the salt water wedge was expected to extend further inland under predevelopment conditions and was therefore more dispersive prior to pumping. Sharp-interface solutions remain useful tools to screen for the vulnerability of coastal aquifers to sea water intrusion, although the significant sources of uncertainty identified in this study require careful consideration to avoid misinterpreting sharp-interface results., (© 2016, National Ground Water Association.)

Objective: The research conducted a large-scale, multisite study on the value and impact of library and information services on patient care., Methods: THE STUDY USED: (1) 2 initial focus groups of librarians; (2) a web-based survey of physicians, residents, and nurses at 56 library sites serving 118 hospitals; and (3) 24 follow-up telephone interviews. Survey respondents were asked to base their responses on a recent incident in which they had sought information for patient care., Results: Of the 16,122 survey respondents, 3/4 said that they had definitely or probably handled aspects of the patient care situation differently as a result of the information. Among the reported changes were advice given to the patient (48%), diagnosis (25%), and choice of drugs (33%), other treatment (31%), and tests (23%). Almost all of the respondents (95%) said the information resulted in a better informed clinical decision. Respondents reported that the information allowed them to avoid the following adverse events: patient misunderstanding of the disease (23%), additional tests (19%), misdiagnosis (13%), adverse drug reactions (13%), medication errors (12%), and patient mortality (6%)., Conclusions: Library and information resources were perceived as valuable, and the information obtained was seen as having an impact on patient care.

Two-component systems mediate bacterial signal transduction, employing a membrane sensor kinase and a cytoplasmic response regulator (RR). Environmental sensing is typically coupled to gene regulation. Understanding how input stimuli activate kinase autophosphorylation remains obscure. The EnvZ/OmpR system regulates expression of outer membrane proteins in response to osmotic stress. To identify EnvZ conformational changes associated with osmosensing, we used HDXMS to probe the effects of osmolytes (NaCl, sucrose) on the cytoplasmic domain of EnvZ (EnvZ(c)). Increasing osmolality decreased deuterium exchange localized to the four-helix bundle containing the autophosphorylation site (His(243)). EnvZ(c) exists as an ensemble of multiple conformations and osmolytes favoured increased helicity. High osmolality increased autophosphorylation of His(243), suggesting that these two events are linked. In-vivo analysis showed that the cytoplasmic domain of EnvZ was sufficient for osmosensing, transmembrane domains were not required. Our results challenge existing claims of robustness in EnvZ/OmpR and support a model where osmolytes promote intrahelical H-bonding enhancing helix stabilization, increasing autophosphorylation and downstream signalling. The model provides a conserved mechanism for signalling proteins that respond to diverse physical and mechanical stimuli.

Human observers can recognize real-world visual scenes with great efficiency. Cortical regions such as the parahippocampal place area (PPA) and retrosplenial complex (RSC) have been implicated in scene recognition, but the specific representations supported by these regions are largely unknown. We used functional magnetic resonance imaging adaptation (fMRIa) and multi-voxel pattern analysis (MVPA) to explore this issue, focusing on whether the PPA and RSC represent scenes in terms of general categories, or as specific scenic exemplars. Subjects were scanned while viewing images drawn from 10 outdoor scene categories in two scan runs and images of 10 familiar landmarks from their home college campus in two scan runs. Analyses of multi-voxel patterns revealed that the PPA and RSC encoded both category and landmark information, with a slight advantage for landmark coding in RSC. fMRIa, on the other hand, revealed a very different picture: both PPA and RSC adapted when landmark information was repeated, but category adaptation was only observed in a small subregion of the left PPA. These inconsistencies between the MVPA and fMRIa data suggests that these two techniques interrogate different aspects of the neuronal code. We propose three hypotheses about the mechanisms that might underlie adaptation and multi-voxel signals., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

In this paper, simple indicators of the propensity for sea water intrusion (SWI) to occur (referred to as "SWI vulnerability indicators") are devised. The analysis is based on an existing analytical solution for the steady-state position of a sharp fresh water-salt water interface. Interface characteristics, that is, the wedge toe location and sea water volume, are used in quantifying SWI in both confined and unconfined aquifers. Rates-of-change (partial derivatives of the analytical solution) in the wedge toe or sea water volume are used to quantify the aquifer vulnerability to various stress situations, including (1) sea-level rise; (2) change in recharge (e.g., due to climate change); and (3) change in seaward discharge. A selection of coastal aquifer cases is used to apply the SWI vulnerability indicators, and the proposed methodology produces interpretations of SWI vulnerability that are broadly consistent with more comprehensive investigations. Several inferences regarding SWI vulnerability arise from the analysis, including: (1) sea-level rise impacts are more extensive in aquifers with head-controlled rather than flux-controlled inland boundaries, whereas the opposite is true for recharge change impacts; (2) sea-level rise does not induce SWI in constant-discharge confined aquifers; (3) SWI vulnerability varies depending on the causal factor, and therefore vulnerability composites are needed that differentiate vulnerability to such threats as sea-level rise, climate change, and changes in seaward groundwater discharge. We contend that the approach is an improvement over existing methods for characterizing SWI vulnerability, because the method has theoretical underpinnings and yet calculations are simple, although the coastal aquifer conceptualization is highly idealized., (© 2011, The Author(s). Ground Water © 2011, National Ground Water Association.)

Spatial navigation is believed to be guided in part by reference to an internal map of the environment. We used functional magnetic resonance imaging (fMRI) to test for a key aspect of a cognitive map: preservation of real-world distance relationships. University students were scanned while viewing photographs of familiar campus landmarks. fMRI response levels in the left hippocampus corresponded to real-world distances between landmarks shown on successive trials, indicating that this region considered closer landmarks to be more representationally similar and more distant landmarks to be more representationally distinct. In contrast, posterior visually responsive regions such as retrosplenial complex and the parahippocampal place area were sensitive to landmark repetition and encoded landmark identity in their multivoxel activity patterns but did not show a distance-related response. These data suggest the existence of a map-like representation in the human medial temporal lobe that encodes the coordinates of familiar locations in large-scale, real-world environments.

In bacterial pathogenesis, virulence gene regulation is controlled by two-component regulatory systems. In Escherichia coli, the EnvZ/OmpR two-component system is best understood as regulating expression of outer membrane proteins, but in Salmonella enterica, OmpR activates transcription of the SsrA/B two-component system located on Salmonella pathogenicity island 2 (SPI-2). The response regulator SsrB controls expression of a type III secretory system in which effectors modify the vacuolar membrane and prevent its degradation via the endocytic pathway. Vacuolar modification enables Salmonella to survive and replicate in the macrophage phagosome and disseminate to the liver and spleen to cause systemic infection. The signals that activate EnvZ and SsrA are unknown but are related to the acidic pH encountered in the vacuole. Our previous work established that SsrB binds to regions of DNA that are AT-rich, with poor sequence conservation. Although SsrB is a major virulence regulator in Salmonella, very little is known regarding how it binds DNA and activates transcription. In the present work, we solved the structure of the C-terminal DNA binding domain of SsrB (SsrB(C)) by NMR and analyzed the effect of amino acid substitutions on function. We identified residues in the DNA recognition helix (Lys(179), Met(186)) and the dimerization interface (Val(197), Leu(201)) that are important for SsrB transcriptional activation and DNA binding. An essential cysteine residue in the N-terminal receiver domain was also identified (Cys(45)), and the effect of Cys(203) on dimerization was evaluated. Our results suggest that although disulfide bond formation is not required for dimerization, dimerization occurs upon DNA binding and is required for subsequent activation of transcription. Disruption of the dimer interface by a C203E substitution reduces SsrB activity. Modification of Cys(203) or Cys(45) may be an important mode of SsrB inactivation inside the host.

Response regulators undergo regulated phosphorylation and dephosphorylation at conserved aspartic acid residues in bacterial signal transduction systems. OmpR is a winged helix-turnhelix DNA-binding protein that functions as a global regulator in bacteria and is also important in pathogenesis. A detailed mechanistic picture of how OmpR binds to DNA and activates transcription is lacking. We used NMR spectroscopy to solve the solution structure of the C-terminal domain of OmpR (OmpR(C)) and to analyze the chemical shift changes that occur upon DNA binding. There is little overlap in the interaction surface with residues of PhoB that were reportedly involved in protein/protein interactions in its head-to-tail dimer. Multiple factors account for the lack of overlap. One is that the spacing between the OmpR half-sites is shorter than observed with PhoB, requiring the arrangement of the two OmpR molecules to be different from that of the PhoB dimer on DNA. A second is the demonstration herein that OmpR can bind to its high affinity site as a monomer. As a result, OmpR(C) appears to be capable of adopting alternative orientations depending on the precise base composition of the binding site, which also contributes to the lack of overlap. In the presence of DNA, chemical shift changes occur in OmpR in the recognition alpha-helix 3, the loop between beta-strand 4 and alpha-helix 1, and the loop between beta-strands 5 and 6. DNA contact residues are Val(203) (T), Arg(207) (G), and Arg(209) (phosphate backbone). Our results suggest that OmpR binds to DNA as a monomer and then forms a symmetric or asymmetric dimer, depending on the binding site. We propose that during activation OmpR binds to DNA and undergoes a conformational change that promotes phosphorylation of the N-terminal receiver domain, the receiver domains dimerize, and then the second monomer binds to DNA. The flexible linker of OmpR enables the second monomer to bind in multiple orientations (head-to-tail and head-to-head), depending on the specific DNA contacts.

Objective: The present experimental study was designed to determine how breastfeeding from a mother who smokes affects infants in the short-term., Methods: Fifteen mother-infant dyads were tested on 2 days separated by 1 week. Mothers smoked (not in the presence of their infants) on one test day and refrained from smoking on the other. For the next 3.5 hours, infants breastfed on demand. Sleep and activity patterns were monitored by placing an actigraph on the infants' leg, and milk intake was determined by weighing the infants before and after each feeding. The nicotine content of the milk was measured to determine the dose of nicotine delivered to the infants., Results: Although there was no significant difference in breast milk intake, despite the taste changes in the milk, infants spent significantly less time sleeping during the hours immediately after their mothers smoked (53.4 minutes), compared with the session when their mothers abstained from smoking (84.5 minutes). This reduction was attributable to shortening of the longest sleep bout and reductions in the amounts of time spent in both active sleep and quiet sleep. With greater doses of nicotine delivered to the infant, less time was spent in active sleep., Conclusions: An acute episode of smoking by lactating mothers altered infants' sleep/wake patterning. Perhaps concerns that their milk would taste like cigarettes and their infants' sleep patterning would be disrupted would motivate lactating mothers to abstain from smoking and to breastfeed longer.

Over the last two decades, changes in technology have allowed academic medical center libraries to bring the world of biomedical information to the physician's computer desktop. Because digital libraries have grown so rapidly and in so many ways, some clinicians may be uncertain about the services and resources that are available to them. This article explains how clinical faculty can best utilize their library to support their research and patient care. It addresses some of the most common myths about the "new" medical library, and it highlights innovations in library resources and services that can help physicians to better access, use and manage medical information.

To provide optimal care for their patients, clinicians must be able to locate and interpret the most current literature. Teaching the necessary skills to medical students is essential, if we wish to train clinicians to be able to keep up with the expansion of biomedical knowledge for their entire working lives. In this paper, we describe our school's four-year curriculum in evidence-based medicine and the performance of three senior classes on the summative evaluation exercise devised to measure the program's success.