scholarly journals Efficient Secretion of Murine IL-2 From an Attenuated Strain of Clostridium sporogenes, a Novel Delivery Vehicle for Cancer Immunotherapy

2021 ◽  
Vol 12 ◽  
Author(s):  
Aleksandra M. Kubiak ◽  
Tom S. Bailey ◽  
Ludwig J. Dubois ◽  
Jan Theys ◽  
Philippe Lambin
Keyword(s):  
Probiotic Strain ◽  
Biologically Active ◽  
Delivery Vehicle ◽  
Clinical Safety ◽  
Clostridium Sporogenes ◽  
Genetically Engineer ◽  
Intratumoral Delivery ◽  
Low Levels ◽  
Spleen And Lymph Nodes ◽  
Close Relatives

Despite a history dating back to the 1800s, using Clostridium bacteria to treat cancer has not advanced beyond the observation that they can colonise and partially destroy solid tumours. Progress has been hampered by their inability to eradicate the viable portion of tumours, and an instinctive anxiety around injecting patients with a bacterium whose close relatives cause tetanus and botulism. However, recent advances in techniques to genetically engineer Clostridium species gives cause to revisit this concept. This paper illustrates these developments through the attenuation of C. sporogenes to enhance its clinical safety, and through the expression and secretion of an immunotherapeutic. An 8.6 kb sequence, corresponding to a haemolysin operon, was deleted from the genome and replaced with a short non-coding sequence. The resultant phenotype of this strain, named C. sporogenes-NT, showed a reduction of haemolysis to levels similar to the probiotic strain, C. butyricum M588. Comparison to the parental strain showed no change in growth or sporulation. Following injection of tumour-bearing mice with purified spores of the attenuated strain, high levels of germination were detected in all tumours. Very low levels of spores and vegetative cells were detected in the spleen and lymph nodes. The new strain was transformed with four different murine IL-2-expressing plasmids, differentiated by promoter and signal peptide sequences. Biologically active mIL-2, recovered from the extracellular fraction of bacterial cultures, was shown to stimulate proliferation of T cells. With this investigation we propose a new, safer candidate for intratumoral delivery of cancer immunotherapeutics.

scholarly journals Mutant PrPSc Conformers Induced by a Synthetic Peptide and Several Prion Strains

2004 ◽  
Vol 78 (4) ◽  
pp. 2088-2099 ◽  
Author(s):  
Patrick Tremblay ◽  
Haydn L. Ball ◽  
Kiyotoshi Kaneko ◽  
Darlene Groth ◽  
Ramanujan S. Hegde ◽  
...  
Keyword(s):  
Prion Disease ◽  
Synthetic Peptide ◽  
Biologically Active ◽  
Spongiform Encephalopathy ◽  
Human Prion Disease ◽  
Prion Strains ◽  
Protease Digestion ◽  
Brain Extracts ◽  
Low Levels ◽  
Prp Gene

ABSTRACT Gerstmann-Sträussler-Scheinker (GSS) disease is a dominantly inherited, human prion disease caused by a mutation in the prion protein (PrP) gene. One mutation causing GSS is P102L, denoted P101L in mouse PrP (MoPrP). In a line of transgenic mice denoted Tg2866, the P101L mutation in MoPrP produced neurodegeneration when expressed at high levels. MoPrPSc(P101L) was detected both by the conformation-dependent immunoassay and after protease digestion at 4°C. Transmission of prions from the brains of Tg2866 mice to those of Tg196 mice expressing low levels of MoPrP(P101L) was accompanied by accumulation of protease-resistant MoPrPSc(P101L) that had previously escaped detection due to its low concentration. This conformer exhibited characteristics similar to those found in brain tissue from GSS patients. Earlier, we demonstrated that a synthetic peptide harboring the P101L mutation and folded into a β-rich conformation initiates GSS in Tg196 mice (29). Here we report that this peptide-induced disease can be serially passaged in Tg196 mice and that the PrP conformers accompanying disease progression are conformationally indistinguishable from MoPrPSc(P101L) found in Tg2866 mice developing spontaneous prion disease. In contrast to GSS prions, the 301V, RML, and 139A prion strains produced large amounts of protease-resistant PrPSc in the brains of Tg196 mice. Our results argue that MoPrPSc(P101L) may exist in at least several different conformations, each of which is biologically active. Such conformations occurred spontaneously in Tg2866 mice expressing high levels of MoPrPC(P101L) as well as in Tg196 mice expressing low levels of MoPrPC(P101L) that were inoculated with brain extracts from ill Tg2866 mice, with a synthetic peptide with the P101L mutation and folded into a β-rich structure, or with prions recovered from sheep with scrapie or cattle with bovine spongiform encephalopathy.

scholarly journals Therapeutic Effects of Probiotic Minas Frescal Cheese on the Attenuation of Ulcerative Colitis in a Murine Model

2021 ◽  
Vol 12 ◽  
Author(s):  
Bárbara F. Cordeiro ◽  
Juliana L. Alves ◽  
Giovanna A. Belo ◽  
Emiliano R. Oliveira ◽  
Marina P. Braga ◽  
...  
Keyword(s):  
Ulcerative Colitis ◽  
Functional Foods ◽  
Activity Index ◽  
Disease Activity Index ◽  
Probiotic Strain ◽  
Human Colon ◽  
Probiotic Bacteria ◽  
Therapeutic Effects ◽  
Probiotic Cheese ◽  
Spleen And Lymph Nodes

Inflammatory bowel diseases (IBDs) constitute disturbances of gastrointestinal tract that cause irreversible changes in the structure and function of tissues. Ulcerative colitis (UC), the most frequent IBD in the population, is characterized by prominent inflammation of the human colon. Functional foods containing probiotic bacteria have been studied as adjuvants to the treatment or prevention of IBDs. The selected probiotic strain Lactococcus lactis NCDO 2118 (L. lactis NCDO 2118) exhibits immunomodulatory effects, with promising results in UC mouse model induced by dextran sodium sulfate (DSS). Additionally, cheese is a dairy food that presents high nutritional value, besides being a good delivery system that can be used to improve survival and enhance the therapeutic effects of probiotic bacteria in the host. Therefore, this work investigated the probiotic therapeutic effects of an experimental Minas Frescal cheese containing L. lactis NCDO 2118 in DSS-induced colitis in mice. During colitis induction, mice that consumed the probiotic cheese exhibited reduced in the severity of colitis, with attenuated weight loss, lower disease activity index, limited shortening of the colon length, and reduced histopathological score. Moreover, probiotic cheese administration increased gene expression of tight junctions’ proteins zo-1, zo-2, ocln, and cln-1 in the colon and increase IL-10 release in the spleen and lymph nodes. In this way, this work demonstrates that consumption of probiotic Minas Frescal cheese, containing L. lactis NCDO 2118, prevents the inflammatory process during DSS-induced colitis in mice, opening perspectives for the development of new probiotic functional foods for personalized nutrition in the context of IBD.

Peripheral metabolism of PTH: fate of biologically active amino terminus in vivo

1988 ◽  
Vol 255 (6) ◽  
pp. E886-E893 ◽  
Author(s):  
F. R. Bringhurst ◽  
A. M. Stern ◽  
M. Yotts ◽  
N. Mizrahi ◽  
G. V. Segre ◽  
...  
Keyword(s):  
Specific Activity ◽  
Limited Proteolysis ◽  
High Specific Activity ◽  
Biologically Active ◽  
Amino Terminal ◽  
Liver And Kidney ◽  
Low Levels ◽  
Peripheral Metabolism

Clearance of intact parathyroid hormone (PTH) from blood is associated with rapid uptake by liver and kidney, limited proteolysis by tissue endopeptidases and, within minutes, appearance of circulating carboxyl-(COOH)-terminal PTH fragments. The fate of the corresponding amino(NH2)-terminal portion of the hormone during this peripheral metabolism is still unknown, however. To determine this, we have employed [35S]bovine PTH (bPTH) labeled to high specific activity at NH2-terminal methionines, which permits direct monitoring of the fate of the PTH NH2-terminus during metabolism in vivo. The [35S]PTH was administered by bolus or continuous intravenous infusion to anesthetized normal rats, to rats subjected to acute ablation of the liver, the kidneys, or both, and to rats receiving co-infusions of excess synthetic bPTH(1-34) NH2-terminal fragments. Analysis by high-resolution chromatographic techniques sensitive to 10(-13) M [35S]PTH peptides in plasma yields no evidence that peripheral metabolism of PTH generates circulating NH2-terminal fragments, even when special measures are taken to block clearance of such putative fragments from blood. We find that the NH2-terminus of PTH is rapidly degraded in situ by the liver but that both liver and especially kidney nevertheless contain low levels of NH2-terminal PTH fragments that, although not released into the blood, are large enough to be potentially active. Thus, the peripheral metabolism of PTH in normal animals does not normally lead to the formation of circulating amino terminal fragments of the hormone that might act independently of intact PTH on peripheral target tissues.

Effect of cigarette smoking on levels of bioavailable testosterone in healthy men

2001 ◽  
Vol 100 (6) ◽  
pp. 661-665 ◽  
Author(s):  
Katherine M. ENGLISH ◽  
Peter J. PUGH ◽  
Helen PARRY ◽  
Nanette E. SCUTT ◽  
Kevin S. CHANNER ◽  
...  
Keyword(s):  
Cigarette Smoking ◽  
Case Control Study ◽  
Free Testosterone ◽  
Early Morning ◽  
Biologically Active ◽  
Sex Hormone Binding Globulin ◽  
Bioavailable Testosterone ◽  
Low Levels ◽  
Never Smokers ◽  
Control Study

The effect of smoking on androgen levels is important given the recent interest in the link between low levels of androgens and the development of cardiovascular disease. Numerous studies examining the effects of cigarette smoking on the levels of total and free testosterone have reported conflicting findings, but there has been no accurate assessment of the effects of cigarette smoking on the levels of bioavailable testosterone [not bound to sex hormone-binding globulin (SHBG)]. We attempted to determine whether smoking affects the level of bioavailable testosterone. We undertook a case-control study of 25 healthy male smokers and 25 healthy never-smokers, matched by age and body mass index. Early morning levels of total, free and bioavailable testosterone, 17β-oestradiol, SHBG and cotinine were determined and compared between the two groups. Levels of total (18.5 ± 4.6 ± nM versus 15.1 ± 4.9 ± nM, P = 0.01) and free testosterone (462 ± 91 ± pM versus 402 ± 93 ± pM, P = 0.03) were found to be higher in smokers compared with non-smokers respectively, as was SHBG (34.1 ± 12.8 versus 28.1 ± 9.0 ± nM, P = 0.06). There were no significant differences in the levels of bioavailable testosterone (3.78 ± 1.59 versus 3.51 ± 1.26 ± nM, P = 0.49) or 17β-oestradiol (44.5 ± 11.4 versus 42.3 ± 11.5 ± pM, P = 0.50) between smokers and non-smokers respectively. These data suggest that cigarette smoking has no significant effect on the biologically active fraction of testosterone, but may influence the levels of total and free testosterone through changes in the levels of SHBG.

scholarly journals Potential of the Probiotic Lactobacillus Plantarum ATCC 14917 Strain to Produce Functional Fermented Pomegranate Juice

Foods ◽  
2018 ◽  
Vol 8 (1) ◽  
pp. 4 ◽  
Author(s):  
Ioanna Mantzourani ◽  
Stavros Kazakos ◽  
Antonia Terpou ◽  
Athanasios Alexopoulos ◽  
Eugenia Bezirtzoglou ◽  
...  
Keyword(s):  
Lactobacillus Plantarum ◽  
Probiotic Strain ◽  
Pomegranate Juice ◽  
Total Phenolic ◽  
Acid Fermentation ◽  
Research Survey ◽  
Trolox Equivalent ◽  
Low Levels ◽  
And Storage ◽  
Fermented Juice

In this research survey the application of probiotic strain Lactobacillus plantarum ATCC 14917 in pomegranate juice fermentation is sought. Pomegranate juice was fermented for 24 h and then it was stored 4 for 4 weeks. Cell viability retained in high levels after the 24 h of fermentation and storage for 4 weeks (above 8.8 log cfu/mL), while fermented pomegranate juice was scored better at the 4th week of storage compared to non-fermented pomegranate juice. The probiotic strain was effective regarding lactic acid fermentation as was proved through sugar and organic acids analysis. Concentration of ethanol was maintained at low levels (0.3–1% v/v). Fermented pomegranate juice contained more and in higher percentages desirable volatile compounds (alcohols, ketones and esters) even at the 4th week of cold storage compared to non-fermented juice. Antioxidant activity (150.63 mg Trolox equivalent (TE)/100 mL at the 2nd week) and total phenolic content (206.46 mg gallic acid equivalents (GAE)/100 mL at the 2nd week) were recorded in higher levels for all the storage time compared to non-fermented juice.

scholarly journals Interactions between Closely Related Bacterial Strains Are Revealed by Deep Transcriptome Sequencing

2015 ◽  
Vol 81 (24) ◽  
pp. 8445-8456 ◽  
Author(s):  
Pedro González-Torres ◽  
Leszek P. Pryszcz ◽  
Fernando Santos ◽  
Manuel Martínez-García ◽  
Toni Gabaldón ◽  
...  
Keyword(s):  
Transcriptome Sequencing ◽  
Halophilic Bacterium ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Bacterial Strains ◽  
Content Type ◽  
Pure Cultures ◽  
Low Levels ◽  
Close Relatives ◽  
Bacterial Assemblages

ABSTRACTComparative genomics, metagenomics, and single-cell technologies have shown that populations of microbial species encompass assemblages of closely related strains. This raises the question of whether individual bacterial lineages respond to the presence of their close relatives by modifying their gene expression or, instead, whether assemblages simply act as the arithmetic addition of their individual components. Here, we took advantage of transcriptome sequencing to address this question. For this, we analyzed the transcriptomes of two closely related strains of the extremely halophilic bacteriumSalinibacter rubergrown axenically and in coculture. These organisms dominate bacterial assemblages in hypersaline environments worldwide. The strains used here cooccurred in the natural environment and are 100% identical in their 16S rRNA genes, and each strain harbors an accessory genome representing 10% of its complete genome. Overall, transcriptomic patterns from pure cultures were very similar for both strains. Expression was detected along practically the whole genome albeit with some genes at low levels. A subset of genes was very highly expressed in both strains, including genes coding for the light-driven proton pump xanthorhodopsin, genes involved in the stress response, and genes coding for transcriptional regulators. Expression differences between pure cultures affected mainly genes involved in environmental sensing. When the strains were grown in coculture, there was a modest but significant change in their individual transcription patterns compared to those in pure culture. Each strain sensed the presence of the other and responded in a specific manner, which points to fine intraspecific transcriptomic modulation.

scholarly journals Identification and enumeration of Clostridium spp. In sufu

2021 ◽  
Vol 233 ◽  
pp. 02035
Author(s):  
Xu Qiong ◽  
Liu Yang ◽  
Yu Yi ◽  
Zhang Nana ◽  
Wang Yue ◽  
...  
Keyword(s):  
Clostridium Botulinum ◽  
High Throughput Sequencing ◽  
Fermented Food ◽  
Clostridium Beijerinckii ◽  
Potential Hazard ◽  
Clostridium Sporogenes ◽  
Clostridium Bifermentans ◽  
Low Levels ◽  
Food Needs ◽  
Clostridium Spp

In this study, Clostridium spp. was counted in 49 Chinese commercial sufu samples, and 25 samples were detected Clostridium in all samples: the detection rate was 51.02%. About 93.87% of the samples contained Clostridium at low levels (log CFU/g＜3.0) and one sample had over 4.12lg CFU/g indicating potential hazard to consumers. The 16S rDNA identification was carried out on the genus Clostridium, and the isolates were confirmed as Clostridium botulinum, Clostridium sporogenes, Clostridium beijerinckii and Clostridium bifermentans, while Clostridium butyricum, Clostridium botulinum, Clostridium perfringens and Clostridium bifermentans were the species detected in the sufu samples by high-throughput sequencing. Based on these results, the research on the number and distribution of clostridium and the possible risks of biotoxins in fermented food needs to be strengthened. To our knowledge, this is the first study to isolate and identify Clostridium spp. in sufu.

scholarly journals Enhancing microRNA activity through increased endosomal release mediated by nigericin

2018 ◽  
Author(s):  
Esteban A. Orellana ◽  
Loganathan Rangasamy ◽  
Srinivasarao Tenneti ◽  
Ahmed M. Abdelaal ◽  
Philip S. Low ◽  
...  
Keyword(s):  
Biologically Active ◽  
Endosomal Escape ◽  
Potassium Ions ◽  
Delivery Vehicle ◽  
Early Endosomes ◽  
Intracellular Compartments ◽  
Delivery Vehicles ◽  
The Difference ◽  
Endosomal Release ◽  
Rnai Response

AbstractThe therapeutic promise of small RNA therapeutics (siRNAs, miRNAs) is not only limited by the lack of delivery vehicles, but also by the inability of the small RNAs to reach intracellular compartments where they can be biologically active. We previously reported successful delivery of functionally active miRNAs via receptor-mediated endocytosis1. This type of targeted therapy still faces one of the major challenges in the delivery field, endosomal sequestration. Here, a new method has been developed to promote endosomal escape of delivered miRNA. The strategy relies on the difference in solute contents between nascent endosomes and the cytoplasm: early endosomes are rich in sodium ions (Na+) while the intracellular fluid is rich is potassium ions (K+). Exploiting this difference through favoring the influx of K+into the endosomes without the exchange for a osmotically active ion (Na+), results in an osmotic differential leading to endosome swelling and bursting. One molecule that is able to exchange K+for an osmotically inactive hydrogen ion is the ionophore nigericin. Via generating an intramolecular miRNA delivery vehicle, containing a ligand, in this case folate, and nigericin we achieve escape of folate-RNA conjugates (e.g. FolamiRs) from their entrapping endosomes into the cytoplasm where they bind the RNA Induced Silencing complex (RISC) and activate the RNAi response.

Low Levels of Pyridoxal 5'-Phosphate in Patients With Cystic Fibrosis

PEDIATRICS ◽  
1986 ◽  
Vol 78 (2) ◽  
pp. 278-282
Author(s):  
Bahjat A. Faraj ◽  
Daniel B. Caplan ◽  
Vernon M. Camp ◽  
Edith Pilzer ◽  
Michael Kutner
Keyword(s):  
Cystic Fibrosis ◽  
Vitamin B6 ◽  
Active Form ◽  
Serum Levels ◽  
Hepatic Disease ◽  
Biologically Active ◽  
Control Group ◽  
Hospitalized Children ◽  
Low Levels

To determine the effect of cystic fibrosis on the regulation of plasma pyridoxal 5'-phosphate (PLP), the biologically active form of vitamin B6, we measured this compound in plasma from 56 patients with cystic fibrosis. The concentration of PLP in plasma was assayed by a radioenzymatic technique. The results of this study showed that PLP concentration was decreased significantly (6.44 ± 5.20 ng/mL, mean ± SD; median 4.45 ng/mL) in patients with cystic fibrosis as compared with a group of hospitalized children with neither cystic fibrosis nor hepatic disease serving as a control group (13.2 ± 5.04 ng/mL, mean ± SD; median 12.5 ng/mL). Additionally, 25% of the population with cystic fibrosis exhibited exceedingly low plasma PLP level (less than 2.75 ng/mL). In patients with cystic fibrosis, significant inverse linear associations were found between plasma PLP and serum levels of SGOT and SGPT (PLP v SGOT: r = -. 60, P < .03; PLP v SGPT: r = -.50, P < .03). This study demonstrated that a deficiency of plasma PLP is a common abnormality in cystic fibrosis and that the low PLP level may be a reflection of impaired vitamin B6 metabolism associated with this disorder.

scholarly journals Exo-Oligosaccharides of Rhizobium sp. Strain NGR234 Are Required for Symbiosis with Various Legumes

2006 ◽  
Vol 188 (17) ◽  
pp. 6168-6178 ◽  
Author(s):  
Christian Staehelin ◽  
Lennart S. Forsberg ◽  
Wim D'Haeze ◽  
Mu-Yun Gao ◽  
Russell W. Carlson ◽  
...  
Keyword(s):  
Biologically Active ◽  
Nodule Formation ◽  
Glycoside Hydrolase Family ◽  
Nitrogen Fixing ◽  
Low Levels ◽  
Culture Supernatants ◽  
Hydrolase Family ◽  
Dependent Mechanism ◽  
Group 1

ABSTRACT Rhizobia are nitrogen-fixing bacteria that establish endosymbiotic associations with legumes. Nodule formation depends on various bacterial carbohydrates, including lipopolysaccharides, K-antigens, and exopolysaccharides (EPS). An acidic EPS from Rhizobium sp. strain NGR234 consists of glucosyl (Glc), galactosyl (Gal), glucuronosyl (GlcA), and 4,6-pyruvylated galactosyl (PvGal) residues with β-1,3, β-1,4, β-1,6, α-1,3, and α-1,4 glycoside linkages. Here we examined the role of NGR234 genes in the synthesis of EPS. Deletions within the exoF, exoL, exoP, exoQ, and exoY genes suppressed accumulation of EPS in bacterial supernatants, a finding that was confirmed by chemical analyses. The data suggest that the repeating subunits of EPS are assembled by an ExoQ/ExoP/ExoF-dependent mechanism, which is related to the Wzy polymerization system of group 1 capsular polysaccharides in Escherichia coli. Mutation of exoK (NGRΩexoK), which encodes a putative glycanase, resulted in the absence of low-molecular-weight forms of EPS. Analysis of the extracellular carbohydrates revealed that NGRΩexoK is unable to accumulate exo-oligosaccharides (EOSs), which are O-acetylated nonasaccharide subunits of EPS having the formula Gal(Glc)5(GlcA)2PvGal. When used as inoculants, both the exo-deficient mutants and NGRΩexoK were unable to form nitrogen-fixing nodules on some hosts (e.g., Albizia lebbeck and Leucaena leucocephala), but they were able to form nitrogen-fixing nodules on other hosts (e.g., Vigna unguiculata). EOSs of the parent strain were biologically active at very low levels (yield in culture supernatants, ∼50 μg per liter). Thus, NGR234 produces symbiotically active EOSs by enzymatic degradation of EPS, using the extracellular endo-β-1,4-glycanase encoded by exoK (glycoside hydrolase family 16). We propose that the derived EOSs (and not EPS) are bacterial components that play a crucial role in nodule formation in various legumes.

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