A Feed Additive Containing Encapsulated 6-Phytase within Recombinant Yarrowia lipolytica Cells Produced by Cultivation on Fat-Containing Waste

Feed phytases are purchased as a dry culture medium of secreting producers, mostly micellar fungi. These preparations are required to withstand heating up to 75–80 °C because they are intended for mixing with feed components with subsequent granulation by spray drying. For this reason, many phytases that have a high specific activity at 37 °C and correspond to the optimal pH of intestinal chyme are not used in practice. A novel expression system allowing accumulation of the phytase from Obesumbacterium proteus within yeast Yarrowia lipolytica was proposed. Encapsulation increases thermal stability of the enzyme from 55 °C up to 70 °C. The obtained preparation exhibited a high impact on the daily weight gain of a weaned mouse model fed a phosphorus-deficient diet at a dosage 165 phytase activity units (FYT)/kg, whereas a commercial phytase preparation—Ladozyme Proxi derived from Aspergillus ficuum—did not improve the daily weight gain even at the dosage of 15,000 FYT/kg

Using ELP Repeats as a Scaffold for De Novo Construction of Gadolinium-Binding Domains within Multifunctional Recombinant Proteins for Targeted Delivery of Gadolinium to Tumour Cells

Three artificial proteins that bind the gadolinium ion (Gd3+) with tumour-specific ligands were de novo engineered and tested as candidate drugs for binary radiotherapy (BRT) and contrast agents for magnetic resonance imaging (MRI). Gd3+-binding modules were derived from calmodulin. They were joined with elastin-like polypeptide (ELP) repeats from human elastin to form the four-centre Gd3+-binding domain (4MBS-domain) that further was combined with F3 peptide (a ligand of nucleolin, a tumour marker) to form the F3-W4 block. The F3-W4 block was taken alone (E2-13W4 protein), as two repeats (E1-W8) and as three repeats (E1-W12). Each protein was supplemented with three copies of the RGD motif (a ligand of integrin αvβ3) and green fluorescent protein (GFP). In contrast to Magnevist (a Gd-containing contrast agent), the proteins exhibited three to four times higher accumulation in U87MG glioma and A375 melanoma cell lines than in normal fibroblasts. The proteins remained for >24 h in tumours induced by Ca755 adenocarcinoma in C57BL/6 mice. They exhibited stability towards blood proteases and only accumulated in the liver and kidney. The technological advantages of using the engineered proteins as a basis for developing efficient and non-toxic agents for early diagnosis of tumours by MRI as well as part of BRT were demonstrated

A Novel Efficient Producer of Human ω-Amidase (Nit2) in Escherichia Coli

Nit2/ω-amidase catalyzes the hydrolysis of α-ketoglutaramate (KGM, the α-keto acid analogue of glutamine) to α-ketoglutarate and ammonia. The enzyme also catalyzes the amide hydrolysis of monoamides of 4- and 5-C-dicarboxylates, including α-ketosuccinamate (KSM, the α-keto acid analogue of asparagine) and succinamate (SM). Here we describe an inexpensive procedure for high-yield expression of human Nit2 (hNit2) in Escherichia coli and purification of the expressed protein. This work includes: 1) the design of a genetic construct (pQE-Nit22) obtained from the previously described construct (pQE-Nit2) by replacing rare codons within an 81 bp-long DNA fragment preferred by E. coli near the translation initiation site; 2) methods for producing and maintaining the pQE-Nit22 construct; 3) purification of recombinant hNit2; and 4) activity measurements of the purified enzyme with KGM and SM. Important features of the hNit2 gene within the pQE-Nit22 construct are: 1) optimized codon composition, 2) the presence of an N-terminus His6 tag immediately after the initiating codon ATG (Met) that permits efficient purification of the end-product on a Ni-NTA-agarose column. We anticipate that the availability of high yield hNit2/ω-amidase will be helpful in elucidating the normal and pathological roles of this enzyme and in the design of specific inhibitors

Efficacy of Synthetic Peptide Corresponding to the ACTH-Like Sequence of Human Immunoglobulin G1 in Experimental Autoimmune Encephalomyelitis

Peptide immunocortin sequence corresponds to the amino acid residues 11–20 of the variable part of human immunoglobulin G1 (IgG1) heavy chain. Since immunocortin was shown previously to inhibit phagocytosis in peritoneal macrophages and ConA-induced T-lymphocytes proliferation in culture, we suggested that immunocortin administering may be of use for patients with self-immune syndrome. Immunocortin in concentration 10 μM inhibited proliferation of both antigen (myelin)-induced and ConA-induced LN lymphocytes isolated from the lymph nodes of Dark Agouti (DA) rats immunized with chorda shear. The biological trials of the synthetic immunocortin were carried out on the DA rats with induced experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. These in vivo experiments have shown that intraperitoneal injections of immunocortin in a daily dosage 100 μg per animal reduced symptoms of EAE in DA rats

Extracting the speed of sound in the strongly interacting matter created in ultrarelativistic lead-lead collisions at the LHC

International audienceUltrarelativistic nuclear collisions create a strongly interacting state of hot and dense quark-gluon matter that exhibits a remarkable collective flow behavior with minimal viscous dissipation. To gain deeper insights into its intrinsic nature and fundamental degrees of freedom, we extracted the speed of sound in this medium created using lead-lead (PbPb) collisions at a center-of-mass energy per nucleon pair of 5.02 TeV. The data were recorded by the CMS experiment at the CERN LHC and correspond to an integrated luminosity of 0.607 nb$^{-1}$. The measurement is performed by studying the multiplicity dependence of the average transverse momentum of charged particles emitted in head-on PbPb collisions. Our findings reveal that the speed of sound in this matter is nearly half the speed of light, with a squared value of 0.241 $\pm$ 0.002 (stat) $\pm$ 0.016 (syst) in natural units. The effective medium temperature, estimated using the mean transverse momentum, is 219 $\pm$ 8 (syst) MeV. The measured squared speed of sound at this temperature aligns precisely with predictions from lattice quantum chromodynamic (QCD) calculations. This result provides a stringent constraint on the equation of state of the created medium and direct evidence for a deconfined QCD phase being attained in relativistic nuclear collisions

Review of searches for vector-like quarks, vector-like leptons, and heavy neutral leptons in proton-proton collisions at s\sqrt{s} = 13 TeV at the CMS experiment

International audienceThe LHC has provided an unprecedented amount of proton-proton collision data, bringing forth exciting opportunities to address fundamental open questions in particle physics. These questions can potentially be answered by performing searches for very rare processes predicted by models that attempt to extend the standard model of particle physics. The data collected by the CMS experiment in 2015-2018 at a center-of-mass energy of 13 TeV help to test the standard model at the highest precision ever and potentially discover new physics. An interesting opportunity is presented by the possibility of new fermions with masses ranging from the MeV to the TeV scale. Such new particles appear in many possible extensions of the standard model and are well motivated theoretically. They may explain the appearance of three generations of leptons and quarks, the mass hierarchy across the generations, and the nonzero neutrino masses. In this report, the status of searches targeting vector-like quarks, vector-like leptons, and heavy neutral leptons at the CMS experiment is discussed. A complete overview of final states is provided together with their complementarity and partial combination. The discovery potential for several of these searches at the High-Luminosity LHC is also discussed

Search for a standard model-like Higgs boson in the mass range between 70 and 110 GeV in the diphoton final state in proton-proton collisions at s\sqrt{s} = 13 TeV

International audienceThe results of a search for a standard model-like Higgs boson decaying into two photons in the mass range between 70 and 110 GeV are presented. The analysis uses the data set collected by the CMS experiment in proton-proton collisions at $\sqrt{s}$ = 13 TeV corresponding to integrated luminosities of 36.3 fb$^{-1}$, 41.5 fb$^{-1}$ and 54.4 fb$^{-1}$ during the 2016, 2017, and 2018 LHC running periods, respectively. No significant excess over the background expectation is observed and 95% confidence level upper limits are set on the product of the cross section and branching fraction for decays of an additional Higgs boson into two photons. The maximum deviation with respect to the background is seen for a mass hypothesis of 95.4 GeV with a local (global) significance of 2.9 (1.3) standard deviations. The observed upper limit ranges from 15 to 73 fb

Search for a standard model-like Higgs boson in the mass range between 70 and 110 GeV in the diphoton final state in proton-proton collisions at s\sqrt{s} = 13 TeV

International audienceThe results of a search for a standard model-like Higgs boson decaying into two photons in the mass range between 70 and 110 GeV are presented. The analysis uses the data set collected by the CMS experiment in proton-proton collisions at $\sqrt{s}$ = 13 TeV corresponding to integrated luminosities of 36.3 fb$^{-1}$, 41.5 fb$^{-1}$ and 54.4 fb$^{-1}$ during the 2016, 2017, and 2018 LHC running periods, respectively. No significant excess over the background expectation is observed and 95% confidence level upper limits are set on the product of the cross section and branching fraction for decays of an additional Higgs boson into two photons. The maximum deviation with respect to the background is seen for a mass hypothesis of 95.4 GeV with a local (global) significance of 2.9 (1.3) standard deviations. The observed upper limit ranges from 15 to 73 fb

Measurement of inclusive and differential cross sections of single top quark production in association with a W boson in proton-proton collisions at s\sqrt{s} = 13.6 TeV

International audienceThe first measurement of the inclusive and normalised differential cross sections of single top quark production in association with a W boson in proton-proton collisions at a centre-of-mass energy of 13.6 TeV is presented. The data were recorded with the CMS detector at the LHC in 2022, and correspond to an integrated luminosity of 34.7 fb$^{-1}$. The analysed events contain one muon and one electron in the final state. For the inclusive measurement, multivariate discriminants exploiting the kinematic properties of the events are used to separate the signal from the dominant top quark-antiquark production background. A cross section of 82.3 $\pm$ 2.1 (stat) ${}^{+9.9}_{-9.7}$ (syst) $\pm$ 3.3 (lumi) pb is obtained, consistent with the predictions of the standard model. A fiducial region is defined according to the detector acceptance to perform the differential measurements. The resulting differential distributions are unfolded to particle level and show good agreement with the predictions at next-to-leading order in perturbative quantum chromodynamics

Search for flavor changing neutral current interactions of the top quark in final states with a photon and additional jets in proton-proton collisions at s\sqrt{s} = 13 TeV

International audienceA search for the production of a top quark in association with a photon and additional jets via flavor changing neutral current interactions is presented. The analysis uses proton-proton collision data recorded by the CMS detector at a center-of-mass energy of 13 TeV, corresponding to an integrated luminosity of 138 fb$^{-1}$. The search is performed by looking for processes where a single top quark is produced in association with a photon, or a pair of top quarks where one of the top quarks decays into a photon and an up or charm quark. Events with an electron or a muon, a photon, one or more jets, and missing transverse momentum are selected. Multivariate analysis techniques are used to discriminate signal and standard model background processes. No significant deviation is observed over the predicted background. Observed (expected) upper limits are set on the branching fractions of top quark decays: $\mathcal{B}$(t$\to$u$\gamma$) $\lt$ 0.95$\times$10$^{-5}$ (1.20$\times$10$^{-5}$) and $\mathcal{B}$(t$\to$c$\gamma$) $\lt$ 1.51$\times$10$^{-5}$ (1.54$\times$10$^{-5}$) at 95% confidence level, assuming a single nonzero coupling at a time. The obtained limit for $\mathcal{B}$(t$\to$u$\gamma$) is similar to the current best limit, while the limit for $\mathcal{B}$(t$\to$c$\gamma$) is significantly tighter than previous results