Determination and localisation of in situ substrate uptake by anaerobic wastewater treatment granular biofilms

2007 ◽  
Vol 55 (8-9) ◽  
pp. 369-376 ◽  
Author(s):  
G. Collins ◽  
T. Mahony ◽  
A.-M. Enright ◽  
A. Gieseke ◽  
D. de Beer ◽  
...  
Keyword(s):  
Wastewater Treatment ◽  
In Situ Hybridisation ◽  
Substrate Uptake ◽  
Methanogenic Activity ◽  
Anaerobic Wastewater Treatment ◽  
Complementary Approach ◽  
Incubation Experiments ◽  
Testing And Measurement

Radiotracer incubation experiments and beta microimaging, along with fluorescent in situ hybridisation (FISH), are proposed as a complementary approach to specific methanogenic activity testing and measurement of in vitro substrate utilisation rates to understand better the ecophysiology of anaerobic granular biofilms from wastewater treatment reactors.

Application of in-situ H2-assisted biogas upgrading in high-rate anaerobic wastewater treatment

2020 ◽  
Vol 299 ◽  
pp. 122598 ◽  
Author(s):  
Heng Xu ◽  
Kaijun Wang ◽  
Xiaoqian Zhang ◽  
Hui Gong ◽  
Yu Xia ◽  
...  
Keyword(s):  
Wastewater Treatment ◽  
High Rate ◽  
Biogas Upgrading ◽  
Anaerobic Wastewater Treatment

Substrate uptake by Gordonia amarae in activated sludge foams by FISH-MAR

2006 ◽  
Vol 54 (1) ◽  
pp. 39-45 ◽  
Author(s):  
E.L. Carr ◽  
K.L. Eales ◽  
R.J. Seviour
Keyword(s):  
Wastewater Treatment ◽  
Activated Sludge ◽  
Wastewater Treatment Plants ◽  
Carbon Sources ◽  
Mycolic Acid ◽  
Substrate Uptake ◽  
Wastewater Treatment Process ◽  
Pure Cultures ◽  
Gordonia Amarae

Gordonia amarae is a right-angled branching filament belonging to the mycolic acid-containing Actinobacteria which is commonly found in many foaming activated sludge wastewater treatment plants. Although studies on different substrates as sole carbon sources by pure cultures of G. amarae have been carried out, none have examined substrate uptake by this organism in situ. Uptake of several hydrophilic and hydrophobic substrates by G. amarae was evaluated in situ using a combination of fluorescence in situ hybridization and microautoradiography. G. amarae could assimilate a range of both hydrophilic and hydrophobic substrates. From the data, G. amarae appears to be physiologically active under aerobic, anaerobic and anoxic condition (NO2 and NO3) for some substrates. This might explain why attempts to control foaming caused by G. amarae using anoxic and anaerobic selectors have been unsuccessful. This study emphasizes that bacteria can behave differently in situ to pure cultures and that it is important to evaluate the in situ physiology of these bacteria if we are to better understand their role in the wastewater treatment process.

Immunological mechanisms to establish embryo tolerance in early bovine pregnancy

2011 ◽  
Vol 23 (5) ◽  
pp. 619 ◽  
Author(s):  
A. E. Groebner ◽  
K. Schulke ◽  
J. C. Schefold ◽  
G. Fusch ◽  
F. Sinowatz ◽  
...  
Keyword(s):  
In Situ Hybridisation ◽  
Immunohistochemical Analysis ◽  
Kynurenine Pathway ◽  
Endometrial Cells ◽  
Maternal Rejection ◽  
Immunological Mechanisms ◽  
Bovine Pregnancy ◽  
Coculture Model

A well-balanced immunological interaction between mother and the semi-allogenic embryo is of particular importance. The objective of the present study was to analyse mechanisms of immune tolerance in bovine pregnancy during peri-implantation. Simmental heifers inseminated with either cryopreserved spermatozoa or seminal plasma were killed 12, 15 or 18 days after oestrus. Uteri were flushed for the recovery of conceptuses and the ipsilateral intercaruncular endometrium was sampled for gene expression analysis. Indoleamine 2,3-dioxygenase (IDO) mRNA, coding for the initial enzyme of the kynurenine pathway, was 18-fold (P < 0.001) more abundant in the endometrium of Day 18 pregnant v. non-pregnant animals. Tandem mass spectrometry revealed a decrease of endometrial l-tryptophan (P = 0.0008), but an increase of l-kynurenine concentration (P = 0.005) from Day 12 to Day 18, suggesting increasing IDO activity (P < 0.03). An in vitro coculture model of endometrial cells showed an induction of IDO expression following interferon-τ exposure primarily in stroma cells, which was confirmed by in situ hybridisation localising IDO mRNA mainly in deep stroma cells. Immunohistochemical analysis revealed fewer CD45-positive leucocytes in the zona basalis of pregnant animals. Elevated IDO activity may reduce the presence of leucocytes in the pregnant endometrium, providing a possible mechanism for protecting the semi-allogenic conceptus from maternal rejection.

scholarly journals Novel lncRNA UPLA1 mediates tumorigenesis and prognosis in lung adenocarcinoma

2020 ◽  
Vol 11 (11) ◽  
Author(s):  
Xiaoyang Han ◽  
Hua Jiang ◽  
Jianni Qi ◽  
Jiamei Li ◽  
Jinghan Yang ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
In Situ Hybridisation ◽  
Vital Role ◽  
Luciferase Reporter ◽  
Fluorescence In Situ Hybridisation ◽  
Biological Functions ◽  
Pulldown Assay

AbstractWith the development of molecular biotechnology and sequencing techniques, long non-coding RNAs (lncRNAs) have been shown to play a vital role in a variety of cancers including lung cancer. In our previous study, we used RNA sequencing and high-content screening proliferation screening data to identify lncRNAs that were significantly associated with tumour biological functions such as LINC01426. Herein, based on previous work, we report a novel lncRNA UPLA1 (upregulation promoting LUAD-associated transcript-1), which has not been explored or reported in any previous studies. Our results showed that UPLA1 is highly expressed and regulates important biological functions in lung adenocarcinoma. In vitro experiments revealed that UPLA1 promoted the migration, invasion, and proliferation abilities, and is related to cell cycle arrest, in lung adenocarcinoma cells. Moreover, the upregulation of UPLA1 significantly improved the growth of tumours in vivo. We identified that UPLA1 was mainly located in the nucleus using fluorescence in situ hybridisation, and that it promoted Wnt/β-catenin signalling by binding to desmoplakin using RNA pulldown assay and mass spectrometry. Additionally, luciferase reporter assay revealed that YY1 is the transcription factor of UPLA1 and suppressed the expression of UPLA1 as a transcriptional inhibitor. This finding provides important evidence regarding the two roles of YY1 in cancer. Furthermore, in situ hybridisation assay results showed that UPLA1 was closely related to the prognosis and tumour, node, metastasis (TNM) stage of lung adenocarcinoma. In summary, our results suggest that the novel lncRNA UPLA1 promotes the progression of lung adenocarcinoma and may be used as a prognostic marker, and thus, has considerable clinical significance.

Methanogenic and sulphate reducing bacterial population levels in a full-scale anaerobic reactor treating pulp and paper industry wastewater using fluorescence in situ hybridisation

2007 ◽  
Vol 55 (10) ◽  
pp. 183-191 ◽  
Author(s):  
O. Ince ◽  
M. Kolukirik ◽  
Z. Cetecioglu ◽  
O. Eyice ◽  
C. Tamerler ◽  
...  
Keyword(s):  
Relative Abundance ◽  
Paper Industry ◽  
In Situ Hybridisation ◽  
Pulp And Paper Industry ◽  
Pulp And Paper ◽  
Fluorescence In Situ Hybridisation ◽  
Methanogenic Activity ◽  
Anaerobic Reactor ◽  
Reactor Height

In this study, specific methanogenic activity (SMA) test and fluorescence in situ hybridisation (FISH) were respectively used to determine acetoclastic methanogenic capacity, and composition and number of methanogenic and sulphate reducing bacterial (SRB) populations within a full scale anaerobic contact reactor treating a pulp and paper industry effluent. The sludge samples were collected from three different heights along the anaerobic reactor having a difficulty of completely stirring. Performance of the anaerobic reactor in terms of COD removal efficiency varied between 47 and 55% at organic loading rates in a range of 1.6–1.8 kg COD m−3 d−1 and methane yield varied between 0.18 and 0.20 m3CH4 kg CODrem−1. The anaerobic reactor was not operated for 2 weeks during the monitoring period. According to SMA test results, potential methane production rate was 276 mL CH4 gVSS−1 d−1 before the off period of the reactor, however it decreased to 159 mL CH4 gVSS−1 d−1 after this period. SMA test and FISH results along the reactor height showed that the acetoclastic methanogenic activity of the sludge samples, the relative abundance of acetoclastic methanogens, hydrogenotrophic methanogens and acetate oxidising SRB decreased as the reactor height increased, however the relative abundance of non-acetate oxidising SRB increased.

Localisation of mRNA for collagenase in osteocytic, bone surface and chondrocytic cells but not osteoclasts

1995 ◽  
Vol 108 (6) ◽  
pp. 2221-2230
Author(s):  
K. Fuller ◽  
T.J. Chambers
Keyword(s):  
Bone Resorption ◽  
Bone Cells ◽  
In Situ Hybridisation ◽  
Cysteine Proteinases ◽  
Bone Surface ◽  
Matrix Metalloproteinase 1 ◽  
Osteoclastic Resorption ◽  
Stimulation Of

Osteoclasts resorb the extracellular matrix of bone by secreting protons and enzymes into a circumpherentially sealed compartment between the osteoclast and the bone surface. Although the lysosomal cysteine proteinases play a major role in matrix degradation by osteoclasts, collagenase (matrix metalloproteinase-1, EC 3.4.24.7) is also required for osteoclastic bone resorption, and may be directly involved in collagen degradation in the hemivacuole. We assessed the effects of inhibitors of cysteine proteinases and collagenase on bone resorption by osteoclasts isolated from rodent bone. We found that while inhibition of cysteine proteinases strongly suppressed osteoclastic resorption, inhibitors of collagenase were without effect on the number, size, or demineralised fringe of excavations. We could find no evidence of expression of mRNA for collagenase in rat osteoclasts by in situ hybridisation, but found that it was expressed by chondrocytes, bone surface cells and osteocytes adjacent to osteoclasts. The distribution of these cells, and the correlation between increased collagenase production and increased stimulation of osteoclastic resorption in vitro by bone cells, suggests that these cells might be involved in the regulation of bone resorption in situ, and that collagenase production might play a role in this process.

Expression and regulation of Lhx6 and Lhx7, a novel subfamily of LIM homeodomain encoding genes, suggests a role in mammalian head development

Development ◽  
1998 ◽  
Vol 125 (11) ◽  
pp. 2063-2074 ◽  
Author(s):  
M. Grigoriou ◽  
A.S. Tucker ◽  
P.T. Sharpe ◽  
V. Pachnis
Keyword(s):  
Basal Forebrain ◽  
In Situ Hybridisation ◽  
Branchial Arch ◽  
Tooth Formation ◽  
Head Development ◽  
Expression Studies ◽  
Overlapping Domains ◽  
Encoding Genes

LIM-homeobox containing (Lhx) genes encode trascriptional regulators which play critical roles in a variety of developmental processes. We have identified two genes belonging to a novel subfamily of mammalian Lhx genes, designated Lhx6 and Lhx7. Whole-mount in situ hybridisation showed that Lhx6 and Lhx7 were expressed during mouse embryogenesis in overlapping domains of the first branchial arch and the basal forebrain. More specifically, expression of Lhx6 and Lhx7 was detected prior to initiation of tooth formation in the presumptive oral and odontogenic mesenchyme of the maxillary and mandibular processes. During tooth formation, expression was restricted to the mesenchyme of individual teeth. Using explant cultures, we have shown that expression of Lhx6 and Lhx7 in mandibular mesenchyme was under the control of signals derived from the overlying epithelium; such signals were absent from the epithelium of the non-odontogenic second branchial arch. Furthermore, expression studies and bead implantation experiments in vitro have provided strong evidence that Fgf8 is primarily responsible for the restricted expression of Lhx6 and Lhx7 in the oral aspect of the maxillary and mandibular processes. In the telencephalon, expression of both genes was predominantly localised in the developing medial ganglionic eminences, flanking a Fgf8-positive midline region. We suggest that Fgf8 and Lhx6 and Lhx7 are key components of signalling cascades which determine morphogenesis and differentiation in the first branchial arch and the basal forebrain.

Molecular cloning, characterisation and expression of two catalase genes from peach

2004 ◽  
Vol 31 (4) ◽  
pp. 349 ◽  
Author(s):  
Francesca Bagnoli ◽  
Susanna Danti ◽  
Valentina Magherini ◽  
Radiana Cozza ◽  
Anna M. Innocenti ◽  
...  
Keyword(s):  
Stress Responses ◽  
Seasonal Changes ◽  
Prunus Persica ◽  
Vascular Tissue ◽  
In Situ Hybridisation ◽  
Leaf Tissue ◽  
Amino Acid Sequences ◽  
Cdna Clones

Two cDNA clones encoding catalase (Cat1 and Cat2) from peach [Prunus persica (L.) Batsch] were identified, that show homologies to other plant catalases. The nucleotide sequences of the two coding regions showed 88% identity to each other. The amino acid sequences predicted from the two full-length clones showed the highest homology to a catalase from cotton and Nicotiana plumbaginifolia L. and included C-terminal tri-peptides typical of those used to target proteins to peroxisomes. Southern hybridisation analysis suggested the existence of two catalase genes in peach. The expression of Cat1 and Cat2 was determined in seeds, vegetative tissue, leaves during the seasonal cycle and in leaves in response to light / dark treatments. Cat1 had high levels of expression only in leaf tissue and was responsive to light and seasonal changes. Cat2 had high levels of expression in in vitro shoots and was also responsive to seasonal changes, but not to light. In situ hybridisations to leaf tissue indicated that the expression of Cat1 was localised mainly in palisade cells, while Cat2 mRNA was present in the vascular tissue. The results of the expression analysis and in situ hybridisation suggest a role for Cat1 in photorespiration and for Cat2 in stress responses.

Macro and micro nutrient limitation of microbial productivity in oligotrophic subtropical Atlantic waters

2008 ◽  
Vol 5 (2) ◽  
pp. 135 ◽  
Author(s):  
Joanna L. Dixon
Keyword(s):  
Primary Production ◽  
Bacterial Production ◽  
Heterotrophic Bacteria ◽  
Total Carbon ◽  
North East ◽  
Incubation Experiments ◽  
Subtropical Oceans ◽  
Marine Primary Production

Environmental context. The subtropical oceans comprise ~70% of the world’s ocean surface and profoundly affect global biogeochemistry and climate. They are characteristically low-nutrient regions, but, owing to their large extent and often rapid nutrient turnover, may contribute to greater than 30% of the total marine primary production. However, there remains long-standing uncertainty as to what individual or combination of resources, e.g. macro (N, P) and micro (trace metals) nutrients, limit or co-limit marine productivity and thus total carbon fixation in these spatially dominant gyre systems. Abstract. The subtropical oceans are characteristically low-nutrient low-chlorophyll regions, but owing to their geographical dominance and rapid nutrient cycling may contribute >30% of the total marine primary production. The present study investigates the addition of P, Fe, Co and Zn on rates of primary production and heterotrophic bacterial production, through a combination of mesoscale in situ (P, and P + Fe) and in vitro (Co or Zn) bioassay incubation experiments. Results from the bioassay incubation experiments suggest that primary production and chlorophyll a biomass are limited by N and P in this oligotrophic region. However, both were increased further after addition of trace metal micronutrients in the order Fe + Zn ≥ Fe + Co > Fe ≈ Co. In contrast, rates of heterotrophic bacterial production did not appear to be P, or significantly, P + Fe limited, although in situ rates did increase during the first 12 h of mesoscale P fertilisation (which were not mirrored in the mesoscale P + Fe addition). The addition of Co to unfertilised waters increased heterotrophic bacterial production and the numbers of heterotrophic bacteria, Prochlorococcus spp. and Synechococcus spp., suggesting Co limitation. Prochlorococcus spp. were the most abundant autotrophs. The highest increases in both heterotrophic and autotrophic carbon assimilation were shown after in vitro addition of either Co or Zn to mesoscale enriched P + Fe waters, suggesting multiple limitation of microbial growth rates in the subtropical oligotrophic north-east Atlantic.

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