Cell densities in the various layers of the rabbit's striate area

1979 ◽  
Vol 156 (3) ◽  
pp. 269-281 ◽  
Author(s):  
Kurt Fleischhauer ◽  
Angelika Vossel
Keyword(s):  
Author(s):  
W. Shain ◽  
H. Ancin ◽  
H.C. Craighead ◽  
M. Isaacson ◽  
L. Kam ◽  
...  

Neural protheses have potential to restore nervous system functions lost by trauma or disease. Nanofabrication extends this approach to implants for stimulating and recording from single or small groups of neurons in the spinal cord and brain; however, tissue compatibility is a major limitation to their practical application. We are using a cell culture method for quantitatively measuring cell attachment to surfaces designed for nanofabricated neural prostheses.Silicon wafer test surfaces composed of 50-μm bars separated by aliphatic regions were fabricated using methods similar to a procedure described by Kleinfeld et al. Test surfaces contained either a single or double positive charge/residue. Cyanine dyes (diIC18(3)) stained the background and cell membranes (Fig 1); however, identification of individual cells at higher densities was difficult (Fig 2). Nuclear staining with acriflavine allowed discrimination of individual cells and permitted automated counting of nuclei using 3-D data sets from the confocal microscope (Fig 3). For cell attachment assays, LRM5 5 astroglial cells and astrocytes in primary cell culture were plated at increasing cell densities on test substrates, incubated for 24 hr, fixed, stained, mounted on coverslips, and imaged with a 10x objective.


2020 ◽  
Vol 640 ◽  
pp. 79-105
Author(s):  
ET Porter ◽  
E Robins ◽  
S Davis ◽  
R Lacouture ◽  
JC Cornwell

Anthropogenic disturbances in the Chesapeake Bay (USA) have depleted eastern oyster Crassostrea virginica abundance and altered the estuary’s environment and water quality. Efforts to rehabilitate oyster populations are underway; however, the effect of oyster biodeposits on water quality and plankton community structure are not clear. In July 2017, we used 6 shear turbulence resuspension mesocosms (STURMs) to determine differences in plankton composition with and without the daily addition of oyster biodeposits to a muddy sediment bottom. STURM systems had a volume-weighted root mean square turbulent velocity of 1.08 cm s-1, energy dissipation rate of ~0.08 cm2 s-3, and bottom shear stress of ~0.36-0.51 Pa during mixing-on periods during 4 wk of tidal resuspension. Phytoplankton increased their chlorophyll a content in their cells in response to low light in tanks with biodeposits. The diatom Skeletonema costatum bloomed and had significantly longer chains in tanks without biodeposits. These tanks also had significantly lower concentrations of total suspended solids, zooplankton carbon, and nitrite +nitrate, and higher phytoplankton carbon concentrations. Results suggest that the absence of biodeposit resuspension initiates nitrogen uptake for diatom reproduction, increasing the cell densities of S. costatum. The low abundance of the zooplankton population in non-biodeposit tanks suggests an inability of zooplankton to graze on S. costatum and negative effects of S. costatum on zooplankton. A high abundance of the copepod Acartia tonsa in biodeposit tanks may have reduced S. costatum chain length. Oyster biodeposit addition and resuspension efficiently transferred phytoplankton carbon to zooplankton carbon, thus supporting the food web in the estuary.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mathias Fink ◽  
Monika Cserjan-Puschmann ◽  
Daniela Reinisch ◽  
Gerald Striedner

AbstractTremendous advancements in cell and protein engineering methodologies and bioinformatics have led to a vast increase in bacterial production clones and recombinant protein variants to be screened and evaluated. Consequently, an urgent need exists for efficient high-throughput (HTP) screening approaches to improve the efficiency in early process development as a basis to speed-up all subsequent steps in the course of process design and engineering. In this study, we selected the BioLector micro-bioreactor (µ-bioreactor) system as an HTP cultivation platform to screen E. coli expression clones producing representative protein candidates for biopharmaceutical applications. We evaluated the extent to which generated clones and condition screening results were transferable and comparable to results from fully controlled bioreactor systems operated in fed-batch mode at moderate or high cell densities. Direct comparison of 22 different production clones showed great transferability. We observed the same growth and expression characteristics, and identical clone rankings except one host-Fab-leader combination. This outcome demonstrates the explanatory power of HTP µ-bioreactor data and the suitability of this platform as a screening tool in upstream development of microbial systems. Fast, reliable, and transferable screening data significantly reduce experiments in fully controlled bioreactor systems and accelerate process development at lower cost.


2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Amber R Paulson ◽  
Maureen O’Callaghan ◽  
Xue-Xian Zhang ◽  
Paul B Rainey ◽  
Mark R H Hurst

Abstract The function of microbes can be inferred from knowledge of genes specifically expressed in natural environments. Here, we report the in vivo transcriptome of the entomopathogenic bacterium Yersinia entomophaga MH96, captured during initial, septicemic, and pre-cadaveric stages of intrahemocoelic infection in Galleria mellonella. A total of 1285 genes were significantly upregulated by MH96 during infection; 829 genes responded to in vivo conditions during at least one stage of infection, 289 responded during two stages of infection, and 167 transcripts responded throughout all three stages of infection compared to in vitro conditions at equivalent cell densities. Genes upregulated during the earliest infection stage included components of the insecticidal toxin complex Yen-TC (chi1, chi2, and yenC1), genes for rearrangement hotspot element containing protein yenC3, cytolethal distending toxin cdtAB, and vegetative insecticidal toxin vip2. Genes more highly expressed throughout the infection cycle included the putative heat-stable enterotoxin yenT and three adhesins (usher-chaperone fimbria, filamentous hemagglutinin, and an AidA-like secreted adhesin). Clustering and functional enrichment of gene expression data also revealed expression of genes encoding type III and VI secretion system-associated effectors. Together these data provide insight into the pathobiology of MH96 and serve as an important resource supporting efforts to identify novel insecticidal agents.


2020 ◽  
Vol 25 (4) ◽  
pp. 417-432
Author(s):  
Hidetoshi Mori ◽  
Jennifer Bolen ◽  
Louis Schuetter ◽  
Pierre Massion ◽  
Clifford C. Hoyt ◽  
...  

AbstractMultiplex immunofluorescence (mIF) allows simultaneous antibody-based detection of multiple markers with a nuclear counterstain on a single tissue section. Recent studies have demonstrated that mIF is becoming an important tool for immune profiling the tumor microenvironment, further advancing our understanding of the interplay between cancer and the immune system, and identifying predictive biomarkers of response to immunotherapy. Expediting mIF discoveries is leading to improved diagnostic panels, whereas it is important that mIF protocols be standardized to facilitate their transition into clinical use. Manual processing of sections for mIF is time consuming and a potential source of variability across numerous samples. To increase reproducibility and throughput we demonstrate the use of an automated slide stainer for mIF incorporating tyramide signal amplification (TSA). We describe two panels aimed at characterizing the tumor immune microenvironment. Panel 1 included CD3, CD20, CD117, FOXP3, Ki67, pancytokeratins (CK), and DAPI, and Panel 2 included CD3, CD8, CD68, PD-1, PD-L1, CK, and DAPI. Primary antibodies were first tested by standard immunohistochemistry and single-plex IF, then multiplex panels were developed and images were obtained using a Vectra 3.0 multispectral imaging system. Various methods for image analysis (identifying cell types, determining cell densities, characterizing cell-cell associations) are outlined. These mIF protocols will be invaluable tools for immune profiling the tumor microenvironment.


Diagnostics ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 388
Author(s):  
Tatsunori Satoh ◽  
Masataka Kikuyama ◽  
Keiko Sasaki ◽  
Hirotoshi Ishiwatari ◽  
Shinya Kawaguchi ◽  
...  

An appropriate diagnosis is required to avoid unnecessary surgery for gallbladder cholesterol polyps (GChPs) and to appropriately treat pedunculated gallbladder carcinomas (GCs). Generally, polyps >10 mm are regarded as surgical candidates. We retrospectively evaluated plain and contrast-enhanced (CE) computed tomography (CT) findings and histopathological features of 11 early GCs and 10 GChPs sized 10–30 mm to differentiate between GC and GChP >10 mm and determine their histopathological background. Patient characteristics, including polyp size, did not significantly differ between groups. All GCs and GChPs were detected on CE-CT; GCs were detected more often than GChPs on plain CT (73% vs 9%; p < 0.01). Sensitivity, specificity, positive and negative predictive values, and diagnostic accuracy for GCs were 73%, 90%, 89%, 75%, and 81%, respectively. On multivariate analysis, lesion detectability on plain CT was independently associated with GCs (odds ratio, 27.1; p = 0.044). Histopathologically, GChPs consisted of adipose tissue. Although larger vessel areas in GCs than in GChPs was not significant (52,737 μm2 vs 31,906 μm2; p = 0.51), cell densities were significantly greater in GCs (0.015/μm2 vs 0.0080/μm2; p < 0.01). Among GPs larger than 10 mm, plain CT could contribute to differentiating GCs from GChPs.


Foods ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 294
Author(s):  
Sana M’hir ◽  
Pasquale Filannino ◽  
Asma Mejri ◽  
Ali Zein Alabiden Tlais ◽  
Raffaella Di Cagno ◽  
...  

This study investigated the fortification of a carob-based kefir-like beverage (KLB) with whey permeate (WP) and oat flour (OF). The response surface method was used to show the effect of WP and OF concentrations on lactic acid bacteria and yeast cell densities, pH, total titratable acidity (TTA), total phenolics content (TCP), DPPH radical scavenging activity, and overall acceptability (OA) in KLB. The statistical design provided thirteen formulations where OF concentration varied from 3% to 5% and WP from 10% to 15%. The enrichment of carob pods decoction with WP and OF had a positive effect on biomass production. Overall fermentation was shown to increase TPC of KLB. Furthermore, OF supplementation led to the higher levels of TPC and antiradical activity. WP negatively affected OA at linear and quadratic levels, whereas no effect of OF was observed at the linear level. The optimum point was found by using WP at 11.51% and OF at 4.77%. Optimized KLB resulted in an enrichment of bioavailable phenolics derivatives and highly digestible proteins.


1999 ◽  
Vol 89 (12) ◽  
pp. 1233-1239 ◽  
Author(s):  
J. A. McGarvey ◽  
T. P. Denny ◽  
M. A. Schell

One susceptible and two resistant cultivars of tomato were tested for differences in infection by Ralstonia solanacearum and for the subsequent multiplication, colonization, and production of the wilt-inducing virulence factor, exopolysaccharide I (EPS I). Bacterial ingress into the taproot was fastest in the susceptible cv. Marion, followed by the resistant cvs. L285 (fivefold slower) and Hawaii 7996 (15-fold slower). Once inside the taproot, R. solanacearum colonized, to some extent, almost all regions of the resistant and susceptible plants. However, colonization occurred sooner in the susceptible than in the resistant cultivars, as measured by viablecell counts of bacteria in the midstems. Rates of multiplication and maximum bacterial cell densities were also greater in the susceptible than in the resistant cultivars. Growth experiments utilizing xylem fluid from infected and uninfected plants indicated that neither antimicrobial activities nor reduced levels of growth-supporting nutrients in the xylem fluids were responsible for the reduced bacterial multiplication in the resistant cultivars. Quantification of EPS I in the infected plants, using an enzyme-linked immunosorbent assay, revealed that the bacterial populations in the susceptible cultivar produced greater amounts of EPS I per plant than those in the resistant cultivars. Immunofluorescence microscopy using antibodies against either EPS I or R. solanacearum cells revealed that bacteria and EPS I were distributed throughout the vascular bundles and intercellular spaces of the pith in the susceptible cultivar, whereas in the resistant cultivars, bacteria and EPS I were restricted to the vascular tissues.


1991 ◽  
Vol 13 (11) ◽  
pp. 873-881 ◽  
Author(s):  
Claudia Käsehagen ◽  
Fritjof Linz ◽  
Gerlinde Kretzmer ◽  
Thomas Scheper ◽  
Karl Schügerl

1972 ◽  
Vol 43 (10) ◽  
pp. 614-622 ◽  
Author(s):  
Lewis Paul Robinson ◽  
Thomas J. De Marco
Keyword(s):  

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