scholarly journals Addition of insoluble fiber to isolation media allows for increased metabolite diversity of lab-cultivable microbes derived from zebrafish gut samples

2019 ◽  
Author(s):  
Alanna R. Condren ◽  
Maria S Costa ◽  
Natalia Rivera Sanchez ◽  
Sindhu Konkapaka ◽  
Kristin L Gallik ◽  
...  
Keyword(s):  
Intestinal Tract ◽  
Optimal Growth ◽  
Bioinformatics Pipeline ◽  
Fecal Matter ◽  
Metabolite Production ◽  
Ph Values ◽  
Diverse Community ◽  
Isolation Media ◽  
Insoluble Fiber

AbstractThere is a gap in measured microbial diversity when comparing genomic sequencing techniques versus cultivation from environmental samples in a laboratory setting. Standardized methods in artificial environments may not recapitulate the environmental conditions that native microbes require for optimal growth. For example, the intestinal tract houses microbes at various pH values as well as minimal oxygen and light environments. These microbes are also exposed to an atypical source of carbon: dietary fiber compacted in fecal matter. To investigate how the addition of insoluble fiber to isolation media could affect the cultivation of microbes from zebrafish intestines, an isolate library was built and analyzed using the bioinformatics pipeline IDBac. The addition of fiber led to an increase in bacterial growth and encouraged the growth of species from several phyla. Furthermore, fiber addition altered the metabolism of the cultivated gut-derived microbes and induced the production of unique metabolites that were not produced when microbes were otherwise grown on standard isolation media. Addition of this inexpensive carbon source to media supported the cultivation of a diverse community whose specialized metabolite production may more closely replicate their metabolite production in vivo.

Activation of isolated heterophils from chicks stimulated in vivo with salmonella enteritidis-immune lymphokines

1996 ◽  
Vol 54 ◽  
pp. 760-761
Author(s):  
R. B. Moyes ◽  
R. E. Droleskey ◽  
M. H. Kogut ◽  
J. R. DeLoach
Keyword(s):  
Lamina Propria ◽  
Intestinal Mucosa ◽  
Salmonella Enteritidis ◽  
Intestinal Tract ◽  
Polymorphonuclear Cells ◽  
Subclinical Infection ◽  
Egg Products ◽  
Immune Lymphokines ◽  
Organ Invasion

Salmonella enteritidis (SE) is of great concern to the poultry industry due to the organism's ability to penetrate the intestinal mucosa of the laying hen and subsequently colonize the ovaries and yolk membrane. The resultant subclinical infection can lead to SE infection of raw eggs and egg products. Interference with the ability of the organism to invade has been linked to the activation and recruitment of inflammatory polymorphonuclear cells, heterophils, to the lamina propria of the intestinal tract.Recently it has been established that heterophil activation and increased resistance to SE organ invasion can be accomplished by the administration of SE-immune lymphokines (SE-ILK) obtained from supernatants of concanavalin-A stimulated SE immune T lymphocytes from SE hyperimmunized hens. Invasion of SE into the lamina propria provides a secondary signal for directing activated heterophils to the site of SE invasion.

scholarly journals Influence of Light Conditions and Medium Composition on Morphophysiological Characteristics of Stevia rebaudiana Bertoni In Vitro and In Vivo

Horticulturae ◽  
2021 ◽  
Vol 7 (7) ◽  
pp. 195
Author(s):  
Alla A. Shulgina ◽  
Elena A. Kalashnikova ◽  
Ivan G. Tarakanov ◽  
Rima N. Kirakosyan ◽  
Mikhail Yu. Cherednichenko ◽  
...  
Keyword(s):  
Culture Media ◽  
Stevia Rebaudiana ◽  
Adventitious Shoot ◽  
Optimal Growth ◽  
Medium Composition ◽  
Light Spectra ◽  
Stevia Rebaudiana Bertoni ◽  
Multiplication Coefficient

We investigated the influence of different conditions (light composition and plant growth regulators (PGRs) in culture media) on the morphophysiological parameters of Stevia rebaudiana Bertoni in vitro and in vivo. Both PGRs and the light spectra applied were found to significantly affect plant morphogenesis. During the micropropagation stage of S. rebaudiana, optimal growth, with a multiplication coefficient of 15, was obtained in an MS culture medium containing 2,4-epibrassinolide (Epin) and indole-3-acetic acid (IAA) at concentrations of 0.1 and 0.5 mg L−1, respectively. During the rooting stage, we found that the addition of 0.5 mg L−1 hydroxycinnamic acid (Zircon) to the MS medium led to an optimal root formation frequency of 85% and resulted in the formation of strong plants with well-developed leaf blades. Cultivation on media containing 0.1 mg L−1 Epin and 0.5 mg L−1 IAA and receiving coherent light irradiation on a weekly basis resulted in a 100% increase in the multiplication coefficient, better adventitious shoot growth, and a 33% increase in the number of leaves. S. rebaudiana microshoots, cultured on MS media containing 1.0 mg L−1 6-benzylaminopurine (BAP) and 0.5 mg L−1 IAA with red monochrome light treatments, increased the multiplication coefficient by 30% compared with controls (white light, media without PGRs).

scholarly journals An optimised GAS-pharyngeal cell biofilm model

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Heema K. N. Vyas ◽  
Jason D. McArthur ◽  
Martina L. Sanderson-Smith
Keyword(s):  
Crystal Violet ◽  
Biofilm Formation ◽  
Cell Model ◽  
Matrix Material ◽  
Three Dimensional ◽  
Growth Period ◽  
Optimal Growth ◽  
Abiotic Surfaces

AbstractGroup A Streptococcus (GAS) causes 700 million infections and accounts for half a million deaths per year. Biofilm formation has been implicated in both pharyngeal and dermal GAS infections. In vitro, plate-based assays have shown that several GAS M-types form biofilms, and multiple GAS virulence factors have been linked to biofilm formation. Although the contributions of these plate-based studies have been valuable, most have failed to mimic the host environment, with many studies utilising abiotic surfaces. GAS is a human specific pathogen, and colonisation and subsequent biofilm formation is likely facilitated by distinct interactions with host tissue surfaces. As such, a host cell-GAS model has been optimised to support and grow GAS biofilms of a variety of GAS M-types. Improvements and adjustments to the crystal violet biofilm biomass assay have also been tailored to reproducibly detect delicate GAS biofilms. We propose 72 h as an optimal growth period for yielding detectable biofilm biomass. GAS biofilms formed are robust and durable, and can be reproducibly assessed via staining/washing intensive assays such as crystal violet with the aid of methanol fixation prior to staining. Lastly, SEM imaging of GAS biofilms formed by this model revealed GAS cocci chains arranged into three-dimensional aggregated structures with EPS matrix material. Taken together, we outline an efficacious GAS biofilm pharyngeal cell model that can support long-term GAS biofilm formation, with biofilms formed closely resembling those seen in vivo.

scholarly journals Brush border protocadherin CDHR2 promotes the elongation and maximized packing of microvilli in vivo

2019 ◽  
Vol 30 (1) ◽  
pp. 108-118 ◽  
Author(s):  
Julia A. Pinette ◽  
Suli Mao ◽  
Bryan A. Millis ◽  
Evan S. Krystofiak ◽  
James J. Faust ◽  
...  
Keyword(s):  
Functional Capacity ◽  
Brush Border ◽  
Intestinal Tract ◽  
Membrane Surface ◽  
Careful Examination ◽  
Tissue Cell ◽  
Apical Surface ◽  
Membrane Surface Area ◽  
Solute Uptake

Transporting epithelial cells optimize their morphology for solute uptake by building an apical specialization: a dense array of microvilli that serves to increase membrane surface area. In the intestinal tract, individual cells build thousands of microvilli, which pack tightly to form the brush border. Recent studies implicate adhesion molecule CDHR2 in the regulation of microvillar packing via the formation of adhesion complexes between the tips of adjacent protrusions. To gain insight on how CDHR2 contributes to brush border morphogenesis and enterocyte function under native in vivo conditions, we generated mice lacking CDHR2 expression in the intestinal tract. Although CDHR2 knockout (KO) mice are viable, body weight trends lower and careful examination of tissue, cell, and brush border morphology revealed several perturbations that likely contribute to reduced functional capacity of KO intestine. In the absence of CDHR2, microvilli are significantly shorter, and exhibit disordered packing and a 30% decrease in packing density. These structural perturbations are linked to decreased levels of key solute processing and transporting factors in the brush border. Thus, CDHR2 functions to elongate microvilli and maximize their numbers on the apical surface, which together serve to increase the functional capacity of enterocyte.

scholarly journals DISCo-microbe: design of an identifiable synthetic community of microbes

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e8534 ◽  
Author(s):  
Dana L. Carper ◽  
Travis J. Lawrence ◽  
Alyssa A. Carrell ◽  
Dale A. Pelletier ◽  
David J. Weston
Keyword(s):  
Microbial Community ◽  
16S Rrna ◽  
Dna Sequence ◽  
Sequence Alignment ◽  
Amplicon Sequencing ◽  
Ribosomal Database Project ◽  
Community Members ◽  
Dna Sequence Alignment ◽  
Diverse Community

Background Microbiomes are extremely important for their host organisms, providing many vital functions and extending their hosts’ phenotypes. Natural studies of host-associated microbiomes can be difficult to interpret due to the high complexity of microbial communities, which hinders our ability to track and identify individual members along with the many factors that structure or perturb those communities. For this reason, researchers have turned to synthetic or constructed communities in which the identities of all members are known. However, due to the lack of tracking methods and the difficulty of creating a more diverse and identifiable community that can be distinguished through next-generation sequencing, most such in vivo studies have used only a few strains. Results To address this issue, we developed DISCo-microbe, a program for the design of an identifiable synthetic community of microbes for use in in vivo experimentation. The program is composed of two modules; (1) create, which allows the user to generate a highly diverse community list from an input DNA sequence alignment using a custom nucleotide distance algorithm, and (2) subsample, which subsamples the community list to either represent a number of grouping variables, including taxonomic proportions, or to reach a user-specified maximum number of community members. As an example, we demonstrate the generation of a synthetic microbial community that can be distinguished through amplicon sequencing. The synthetic microbial community in this example consisted of 2,122 members from a starting DNA sequence alignment of 10,000 16S rRNA sequences from the Ribosomal Database Project. We generated simulated Illumina sequencing data from the constructed community and demonstrate that DISCo-microbe is capable of designing diverse communities with members distinguishable by amplicon sequencing. Using the simulated data we were able to recover sequences from between 97–100% of community members using two different post-processing workflows. Furthermore, 97–99% of sequences were assigned to a community member with zero sequences being misidentified. We then subsampled the community list using taxonomic proportions to mimic a natural plant host–associated microbiome, ultimately yielding a diverse community of 784 members. Conclusions DISCo-microbe can create a highly diverse community list of microbes that can be distinguished through 16S rRNA gene sequencing, and has the ability to subsample (i.e., design) the community for the desired number of members and taxonomic proportions. Although developed for bacteria, the program allows for any alignment input from any taxonomic group, making it broadly applicable. The software and data are freely available from GitHub (https://github.com/dlcarper/DISCo-microbe) and Python Package Index (PYPI).

scholarly journals Inactivation of tyrosine aminotransferase in neutral homogenates and rat liver slices

1972 ◽  
Vol 129 (5) ◽  
pp. 1131-1138 ◽  
Author(s):  
F. Auricchio ◽  
L. Mollica ◽  
A. Liguori
Keyword(s):  
Amino Acid ◽  
Rat Liver ◽  
Acid Concentration ◽  
Tyrosine Aminotransferase ◽  
Amino Acid Concentration ◽  
Acidic Ph ◽  
Ph Values ◽  
Neutral Ph ◽  
Liver Slices

Inactivation of tyrosine aminotransferase induced in vivo by triamcinolone was studied in a homogenate incubated at neutral pH values. The integrity and the presence of subcellular particles together with a compartment of acidic pH are necessary for inactivation of tyrosine aminotransferase. It is suggested that tyrosine aminotransferase is inactivated inside lysosomes. The system responsible for inactivation of tyrosine aminotransferase was partially purified and identified with lysosomal cathepsins B and B1. Inactivation of tyrosine aminotransferase in liver slices is controlled by the amino acid concentration and strongly stimulated by cysteine. 3,3′,5-Tri-iodo-l-thyronine reversibly and strongly decreases the rate of inactivation of tyrosine aminotransferase. The effect is not due to an increased rate of tyrosine aminotransferase synthesis.

Evaluation of a system for intragastric pH monitoring of intensive care unit patients: preliminary report

1992 ◽  
Vol 1 (1) ◽  
pp. 81-84 ◽  
Author(s):  
JM Clochesy
Keyword(s):  
Ph Monitoring ◽  
Gastric Ph ◽  
Gastric Aspirate ◽  
Ph Probe ◽  
Buffer Solutions ◽  
Ph Values ◽  
Small Series ◽  
Significant Difference

BACKGROUND: A new pH probe-tipped nasogastric sump tube is available to monitor gastric pH conveniently. This study assesses its ability to measure gastric acidity accurately. METHODS: The accuracy of the combined pH probe nasogastric tube (GrapHprobe ST) was determined by comparing it with standard buffer solutions (pH 1.0, 2.0, 4.0 and 7.0) traceable to the National Institute of Standards and Technology. Gastric pH values obtained were compared with values obtained using indicator paper and a calibrated glass electrode on gastric aspirate. RESULTS: Although statistically significant differences were found in vitro between the pH of three of the buffer solutions and the pH values obtained by the nasogastric sump tube, the results were within 0.5 pH unit. When rounded to the nearest pH unit, all values were the same as the buffer solutions. No significant difference was found in the pH values obtained during in vivo testing. CONCLUSIONS: The GrapHprobe ST measured gastric pH within reasonable accuracy in this small series.

scholarly journals Early pH Changes in Musculoskeletal Tissues upon Injury—Aerobic Catabolic Pathway Activity Linked to Inter-Individual Differences in Local pH

2020 ◽  
Vol 21 (7) ◽  
pp. 2513 ◽  
Author(s):  
Julia C. Berkmann ◽  
Aaron X. Herrera Martin ◽  
Agnes Ellinghaus ◽  
Claudia Schlundt ◽  
Hanna Schell ◽  
...  
Keyword(s):  
Individual Differences ◽  
Animal Models ◽  
Time Course ◽  
Tricarboxylic Acid Cycle ◽  
Controlled Drug Release ◽  
Post Injury ◽  
Ph Changes ◽  
Ph Values ◽  
Local Ph

Local pH is stated to acidify after bone fracture. However, the time course and degree of acidification remain unknown. Whether the acidification pattern within a fracture hematoma is applicable to adjacent muscle hematoma or is exclusive to this regenerative tissue has not been studied to date. Thus, in this study, we aimed to unravel the extent and pattern of acidification in vivo during the early phase post musculoskeletal injury. Local pH changes after fracture and muscle trauma were measured simultaneously in two pre-clinical animal models (sheep/rats) immediately after and up to 48 h post injury. The rat fracture hematoma was further analyzed histologically and metabolomically. In vivo pH measurements in bone and muscle hematoma revealed a local acidification in both animal models, yielding mean pH values in rats of 6.69 and 6.89, with pronounced intra- and inter-individual differences. The metabolomic analysis of the hematomas indicated a link between reduction in tricarboxylic acid cycle activity and pH, thus, metabolic activity within the injured tissues could be causative for the different pH values. The significant acidification within the early musculoskeletal hematoma could enable the employment of the pH for novel, sought-after treatments that allow for spatially and temporally controlled drug release.

ISOMERIZATION OF 11-cis VITAMIN A IN VIVO

1960 ◽  
Vol 38 (11) ◽  
pp. 1219-1222 ◽  
Author(s):  
D. W. Stainer ◽  
T. K. Murray ◽  
J. A. Campbell
Keyword(s):  
Vitamin A ◽  
Intestinal Tract ◽  
Biological Potency ◽  
Oral Doses ◽  
Poor Absorption ◽  
Cis Isomer

Single oral doses of all-trans and 11-cis vitamin A acetate were given to young, vitamin A deficient rats and the proportion of cis isomer in the intestinal tract and liver measured. Some conversion of 11-cis to all-trans occurred in the stomach and intestine, and a mixture of the two isomers was absorbed and stored in the liver. The high proportion of cis isomer found in the liver stores 5 hours after a dose of 11-cis vitamin A disappeared completely in 23 days. Oral doses of both all-trans and 11-cis vitamin A produced greater liver stores than the same doses given subcutaneously. The relative biological potency of the 11-cis isomer was the same by either route, which indicated that the low potency of this isomer was not due only to poor absorption from the intestine.

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