Diffusion through Pig Gastric Mucin: Effect of Relative Humidity.
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ABSTRACT:
SUBMITTER: Runnsjo A
PROVIDER: S-EPMC4918968 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Mucin glycoproteins consist of tandem-repeating glycosylated regions flanked by non-repetitive protein domains with little glycosylation. These non-repetitive domains are involved in polymerization of mucin and play an important role in the pH-dependent gelation of gastric mucin, which is essential for protecting the stomach from autodigestion. We examine folding of the non-repetitive sequence of PGM-2X (242 amino acids) and the von Willebrand factor vWF-C1 domain (67 amino acids) at neutral and low pH using discrete molecular dynamics (DMD) in an implicit solvent combined with a four-bead peptide model. Using the same implicit solvent parameters, folding of both domains is simulated at neutral and low pH. In contrast to vWF-C1, PGM-2X folding is strongly affected by pH as indicated by changes in the contact order, radius of gyration, free-energy landscape, and the secondary structure. Whereas the free-energy landscape of vWF-C1 shows a single minimum at both neutral and low pH, the free-energy landscape of PGM-2X is characterized by multiple minima that are more numerous and shallower at low pH. Detailed structural analysis shows that PGM-2X partially unfolds at low pH. This partial unfolding is facilitated by the C-terminal region GLU236-PRO242, which loses contact with the rest of the domain due to effective "mean-field" repulsion among highly positively charged N- and C-terminal regions. Consequently, at low pH, hydrophobic amino acids are more exposed to the solvent. In vWF-C1, low pH induces some structural changes, including an increased exposure of CYS at position 67, but these changes are small compared to those found in PGM-2X. For PGM-2X, the DMD-derived average ?-strand propensity increases from 0.26 ± 0.01 at neutral pH to 0.38 ± 0.01 at low pH. For vWF-C1, the DMD-derived average ?-strand propensity is 0.32 ± 0.02 at neutral pH and 0.35 ± 0.02 at low pH. The DMD-derived structural information provides insight into pH-induced changes in the folding of two distinct mucin domains and suggests plausible mechanisms of the aggregation/gelation of mucin.
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Project description:Relative humidity influences microbial communities
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Project description:Polyclonal antibodies raised to deglycosylated pig gastric mucin were used to screen a cDNA library constructed with pig stomach mucosal mRNA. Immunocytochemistry indicated that the antibody recognizes intracellular and secreted mucin in surface mucous cells of pig gastric epithelium. A total of 70 clones producing proteins immunoreactive to this antibody were identified, two of which (PGM-2A,9B) were fully sequenced from both ends. Clone PGM-9B hybridized to a polydisperse mRNA (3-9 kb) from pig stomach, but not liver, intestine or spleen, nor to mRNA from human, mouse, rabbit or rat stomach. Sequence analysis indicated that PGM-9B encodes 33 tandem repeats of a 16-amino-acid consensus sequence rich in serine (46%) and threonine (17%). Using the restriction enzyme MwoI, which has a single target site in the repeat, it was demonstrated that PGM-9B consists entirely of this tandem repeat. Southern-blot analysis indicated that the repeat region is contained in a 20 kb HindIII-EcoRI fragment, and BamHI digestion suggested that most of the repeats are contained in a 10 kb fragment. In situ hybridization with an antisense probe to PGM-9B showed an intense signal in the entire gastric gland. Clone PGM-2A also contains the same repeat sequence as 9B, but, in addition, has a 64-amino-acid-long non-repeat region at its 5' end. Interestingly the non-repeat region of PGM-2A has five cysteine residues, the arrangement of which is identical with that reported for human intestinal mucin gene MUC2.
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Project description:Meteorological parameters are important factors that have an influence on infectious diseases. The present study aimed to explore the correlation between the spread of COVID-19, temperature, and relative humidity. The effect of human-imposed control parameters in the form of lockdown on the dissipation of COVID-19 was also analysed. Data were collected on the three study variables - temperature, relative humidity, and lockdown period - from nine of the most infected cities worldwide as well as information on changes in the number of COVID-19 patients from the beginning to a specific point in the lockdown period. A generalised regression model was applied to explore the effect of temperature and relative humidity on the change in daily new cases of COVID-19. The regression analysis did not find any significant correlation between temperature, humidity, and change in number of COVID-19 cases. Analysis of the cities with wide-ranging temperature variations showed a negative correlation of COVID-19 transmission (P=0.079) with temperature, but a relatively non-significant correlation with relative humidity (P=0.198). The number of total deaths was also higher in low-temperature countries compared with high-temperature countries. The specific growth rate in COVID-19 cases was decreased by more than 66% after implementation of a lockdown. This growth rate was exponentially decreased over time through the proper implementation of lockdown. Analysis of the real-case scenario and application of predictive models showed that for New York, Lombardy, and Madrid more than 120 days of strict lockdown was required for complete control of the transmission of COVID-19.
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Project description:Relative humidity (RH) was measured at hourly intervals for approximately one year in two caves at seven stations near Playa del Carmen in Quintana Roo, Mexico. Sistema Muévelo Rico is a 1.1 km long cave with 12 entrances and almost no dark zone. Río Secreto (Tuch) is a large river cave with more than 40 km of passages, and an extensive dark zone. Given the need for cave specialists to adapt to saturated humidity, presumably by cuticular thinning, the major stress of RH would be its deviation from saturation. RH in Río Secreto (Tuch) was invariant at three sites and displayed short deviations from 100% RH at the other four sites. These deviations were concentrated at the end of the nortes and beginning of the rainy season. Three of the sites in Sistema Muévelo Rico showed a similar pattern although the timing of the deviations from 100% RH was somewhat displaced. Four sites in Sistema Muévelo Rico were more variable, and were analyzed using a measure of amount of time of deviation from 100% RH for each 24 hour period. Strong seasonality was evident but, remarkably, periods of constant high humidity were not the same at all sites. In most Sistema Muévelo Rico sites, there was a detectable 24 hour cycle in RH, although it was quite weak in about half of them. For Río Secreto (Tuch) only one site showed any sign of a 24 hour cycle. The troglomorphic fauna was more or less uniformly spread throughout the caves and did not concentrate in any one area or set of RH conditions. Compared to temperature, RH is much more constant, perhaps even more constant than the amount of light. However, changes in RH as a result of global warming may have a major negative effect on the subterranean fauna.
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Project description:The studies expounding on the effects of storage conditions on texture changes are limited. The researchers have been proposing methods to measure pecan texture instrumentally. But current protocols and/or attributes fail to address huge variability during experimentation. Additionally, there are no predictive models to estimate changes in pecan texture during storage. This study addresses all the above concerns and investigates the effects of different relative humidity (RH, 30-90%) and packaging material (Polyethylene-Nylon [PEN], polypropylene [PP], low density polyethylene [LDPE], and metallic laminates [ML]) on pecan texture, introducing a rift ratio (F/H or fracturability to hardness ratio) to address variability in the data and predictive model to estimate changes in the textural attribute of pecans during storage. The textural analysis was conducted on pecan cores and intact pecans to measure the area under curve, fracturability, hardness, cohesiveness, chewiness, springiness, and rift ratio. It was observed that values for the rift ratio obtained using the intact pecan method had high R2 (0.72) as compared to the rest of the textural attributes. A three-parameter logistic model was employed to predict pecan texture during storage. The pecans stored at 75, 80, and 90% reached the rift ratio (F/H) of 0.5 at approx. 115, 3, and 0.15 days (~ 4 hr), respectively. Similarly, pecans stored in LDPE, PP, and PEN packs at 80% reached rift ratio (F/H) of 0.5 at approx. 26, 57, and 78 days, respectively. The presence of any kind of package delayed fracturability loss by at least eight folds at 80% RH. The pecans stored in ML did not experience a significant change in textural attributes.
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Project description:The effect of acidity and relative humidity on bulk isoprene aerosol parameters has been investigated in several studies; however, few measurements have been conducted on individual aerosol compounds. The focus of this study has been the examination of the effect of acidity and relative humidity on secondary organic aerosol (SOA) chemical composition from isoprene photooxidation in the presence of nitrogen oxide (NO x ). A detailed characterization of SOA at the molecular level was also investigated. Experiments were conducted in a 14.5 m3 smog chamber operated in flow mode. Based on a detailed analysis of mass spectra obtained from gas chromatography-mass spectrometry of silylated derivatives in electron impact and chemical ionization modes, ultra-high performance liquid chromatography/electrospray ionization/time-of-flight high-resolution mass spectrometry, and collision-induced dissociation in the negative ionization modes, we characterized not only typical isoprene products but also new oxygenated compounds. A series of nitroxy-organosulfates (NOSs) were tentatively identified on the basis of high-resolution mass spectra. Under acidic conditions, the major identified compounds include 2-methyltetrols (2MT), 2-methylglyceric acid (2mGA), and 2MT-OS. Other products identified include epoxydiols, mono- and dicarboxylic acids, other organic sulfates, and nitroxy- and nitrosoxy-OS. The contribution of SOA products from isoprene oxidation to PM2.5 was investigated by analyzing ambient aerosol collected at rural sites in Poland. Methyltetrols, 2mGA, and several organosulfates and nitroxy-OS were detected in both the field and laboratory samples. The influence of relative humidity on SOA formation was modest in non-acidic-seed experiments and stronger under acidic seed aerosol. Total secondary organic carbon decreased with increasing relative humidity under both acidic and non-acidic conditions. While the yields of some of the specific organic compounds decreased with increasing relative humidity, others varied in an indeterminate manner from changes in the relative humidity.
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Project description:Pig gastric mucins were isolated from the surface epithelium of the cardiac region, corpus and antrum using density-gradient centrifugation after extraction in 6 M guanidinium chloride. In CsCl/0.5 M guanidinium chloride, mucins solubilized from the cardiac region appeared as a broad unimodal band at 1.52 g/ml whereas those from the corpus and antrum occurred as high- and low-density populations at 1.50 and 1.45 g/ml respectively. High-iron diamine reacted more strongly with the cardiac mucins and the high-density populations from corpus and antrum than with the two low-density ones. In keeping with this, approx. 60% of the oligosaccharides from the former mucins and 20% from the latter contained sulphate. All surface epithelial cells of the cardiac region stained with high-iron diamine, whereas in the corpus only the epithelium in the bottom of the pits reacted, suggesting that the high-density population from this region originates from these cells. Mucins from all regions were composed of subunits, each containing highly glycosylated domains. The mucins from the cardiac region were larger than those from the corpus and antrum, and reduced subunits as well as high-molecular-mass glycopeptides from the cardiac mucins were larger than the corresponding fragments from the other regions. Ion-exchange HPLC showed that reduced subunits from the cardiac mucins and the high-density populations from the corpus and antrum were more 'acidic' than reduced subunits from the two low-density ones. All mucins contained a 'neutral'fraction, in particular those from the antrum. Pig gastric mucus thus contains a number of distinctly different mucin populations varying in buoyant density, size, 'acidity', glycosylation, sulphation and tissue origin.
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Project description:The survival on stainless steel of ten Salmonella isolates from food factory, clinical and veterinary sources was investigated. Stainless steel coupons inoculated with Salmonella were dried and stored at a range of temperatures and relative humidity (RH) levels representing factory conditions. Viability was determined from 1 to 22 days. Survival curves obtained for most isolates and storage conditions displayed exponential inactivation described by a log-linear model. Survival was affected by environmental temperatures and RH with decimal reduction times (DRTs) ranging from <1 day to 18 days. At 25 °C/15% RH, all isolates survived at levels of 103 to 105 cfu for >22 days. Furthermore, temperatures and RH independently influenced survival on stainless steel; increasing temperatures between 10 °C and 37 °C and increasing RH levels from 30-70% both decreased the DRT values. Survival curves displaying a shoulder followed by exponential death were obtained for three isolates at 10 °C/70% RH. Inactivation kinetics for these were described by modified Weibull models, suggesting that cumulative injury occurs before cellular inactivation. This study highlights the need to control temperature and RH to limit microbial persistence in the food manufacturing environment, particularly during the factory shut-down period for cleaning when higher temperature/humidity levels could be introduced.
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