Development and Evaluation of a New Lateral Flow Immunoassay for Serodiagnosis of Human Fasciolosis

Fasciolosis is an important plant-borne trematode zoonosis. This disease is of both clinical and veterinary relevance and, according to the WHO, is considered a re-emerging disease that is spreading around the world. Fasciolosis has a serious impact on health because of the large size of the parasite and the effects of the parasite in down-regulating the host immune response. Human fasciolosis can be distinguished by an acute phase, in which the parasite migrates through different tissues, and a chronic phase in which it invades the bile ducts. Here we describe the development of a rapid, simple and inexpensive immunochromatographic diagnostic method, based on the use of a recombinant cathepsin L1 protein, which performs better than other more complex indirect methods, providing similar specificity and higher sensitivity. The simplicity of the method represents a great advantage for the intervention systems applied in different endemic areas by WHO, such as passive case finding (e.g. Vietnam) and selective treatment (e.g. Egypt). Because of its characteristics, the system can be applied to both phases of the disease, and in holo, meso and hyperendemic areas where point-of-care testing is required

Detection of plasmin based on specific peptide substrate using acoustic transducer

© 2015 Elsevier B.V. All rights reserved. In this work we report the detection of plasmin protease by means of the thickness shear mode (TSM) acoustic method. The biorecognition element consists of a peptide substrate (PS) specific to plasmin immobilized on a piezoelectric quartz crystal electrode. After enzymatic reaction with plasmin, it cleaves a short fragment of the peptide causing increase in the resonance frequency of the piezo crystal. Plasmin was detected in the range of concentrations 1-20 nM, a target interval in which its presence presumably affects the quality of milk. The PS exhibited negligible response against to similar protease trypsin. This has been confirmed also by electrochemical detection method. Limit of detection of this acoustic transducer was found to be 0.65 nM. Formation of the sensing surface and kinetic effect of plasmin on the peptide substrate was studied by atomic force microscopy (AFM). The PS response was also validated in pretreated milk samples spiked by known concentrations of plasmin achieving an average recovery of 63 ± 0.6%

Counting the number of τ-exceptional sequences over Nakayama algebras

The notion of a τ-exceptional sequence was introduced by Buan and Marsh in (2018) as a generalisation of an exceptional sequence for finite dimensional algebras. We calculate the number of complete τ-exceptional sequences over certain classes of Nakayama algebras. In some cases, we obtain closed formulas which also count other well known combinatorial objects, and exceptional sequences of path algebras of Dynkin quivers

Neonatal Fc Receptor: From Immunity to Therapeutics

The neonatal Fc receptor (FcRn), also known as the Brambell receptor and encoded by Fcgrt, is a MHC class I like molecule that functions to protect IgG and albumin from catabolism, mediates transport of IgG across epithelial cells, and is involved in antigen presentation by professional antigen presenting cells. Its function is evident in early life in the transport of IgG from mother to fetus and neonate for passive immunity and later in the development of adaptive immunity and other functions throughout life. The unique ability of this receptor to prolong the half-life of IgG and albumin has guided engineering of novel therapeutics. Here, we aim to summarize the basic understanding of FcRn biology, its functions in various organs, and the therapeutic design of antibody- and albumin-based therapeutics in light of their interactions with FcRn

The recycling and transcytotic pathways for IgG transport by FcRn are distinct and display an inherent polarity

The Fc receptor FcRn traffics immunoglobulin G (IgG) in both directions across polarized epithelial cells that line mucosal surfaces, contributing to host defense. We show that FcRn traffics IgG from either apical or basolateral membranes into the recycling endosome (RE), after which the actin motor myosin Vb and the GTPase Rab25 regulate a sorting step that specifies transcytosis without affecting recycling. Another regulatory component of the RE, Rab11a, is dispensable for transcytosis, but regulates recycling to the basolateral membrane only. None of these proteins affect FcRn trafficking away from lysosomes. Thus, FcRn transcytotic and recycling sorting steps are distinct. These results are consistent with a single structurally and functionally heterogeneous RE compartment that traffics FcRn to both cell surfaces while discriminating between recycling and transcytosis pathways polarized in their direction of transport

Conjugation of a Ru(II) Arene Complex to Neomycin or to Guanidinoneomycin Leads to Compounds with Differential Cytotoxicities and Accumulation between Cancer and Normal Cells

A straightforward methodology for the synthesis of conjugates between a cytotoxic organometallic ruthenium(II) complex and amino- and guanidinoglycosides, as potential RNA-targeted anticancer compounds, is described. Under microwave irradiation, the imidazole ligand incorporated on the aminoglycoside moiety (neamine or neomycin) was found to replace one triphenylphosphine ligand from the ruthenium precursor [(η6-p-cym)RuCl(PPh3)2]+, allowing the assembly of the target conjugates. The guanidinylated analogue was easily prepared from the neomycin-ruthenium conjugate by reaction with N,N′-di-Boc-N″-triflylguanidine, a powerful guanidinylating reagent that was compatible with the integrity of the metal complex. All conjugates were purified by semipreparative high-performance liquid chromatography (HPLC) and characterized by electrospray ionization (ESI) and matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) and NMR spectroscopy. The cytotoxicity of the compounds was tested in MCF-7 (breast) and DU-145 (prostate) human cancer cells, as well as in the normal HEK293 (Human Embryonic Kidney) cell line, revealing a dependence on the nature of the glycoside moiety and the type of cell (cancer or healthy). Indeed, the neomycin-ruthenium conjugate (2) displayed moderate antiproliferative activity in both cancer cell lines (IC50 ≈ 80 μM), whereas the neamine conjugate (4) was inactive (IC50 ≈ 200 μM). However, the guanidinylated analogue of the neomycin-ruthenium conjugate (3) required much lower concentrations than the parent conjugate for equal effect (IC50 = 7.17 μM in DU-145 and IC50 = 11.33 μM in MCF-7). Although the same ranking in antiproliferative activity was found in the nontumorigenic cell line (3 2 > 4), IC50 values indicate that aminoglycoside-containing conjugates are about 2-fold more cytotoxic in normal cells (e.g., IC50 = 49.4 μM for 2) than in cancer cells, whereas an opposite tendency was found with the guanidinylated conjugate, since its cytotoxicity in the normal cell line (IC50 = 12.75 μM for 3) was similar or even lower than that found in MCF-7 and DU-145 cancer cell lines, respectively. Cell uptake studies performed by ICP-MS with conjugates 2 and 3 revealed that guanidinylation of the neomycin moiety had a positive effect on accumulation (about 3-fold higher in DU-145 and 4-fold higher in HEK293), which correlates well with the higher antiproliferative activity of 3. Interestingly, despite the slightly higher accumulation in the normal cell than in the cancer cell line (about 1.4-fold), guanidinoneomycin-ruthenium conjugate (3) was more cytotoxic to cancer cells (about 1.8-fold), whereas the opposite tendency applied for neomycin-ruthenium conjugate (2). Such differences in cytotoxic activity and cellular accumulation between cancer and normal cells open the way to the creation of more selective, less toxic anticancer metallodrugs by conjugating cytotoxic metal-based complexes such as ruthenium(II) arene derivatives to guanidinoglycosides

Luteinizing hormone-releasing hormone analogs with increased anti-ovulatory activity

A series of LH-RH antagonist analogs has been developed in which inhibitory activities have been increased to a potentially clinically useful level. The new peptides, which are typified by [N-acetyl-D-p-Cl-Phe 1,2, D-Trp 3, D-Phe 6,D-Ala 10]-LH-RH and [N-acetyl-D-Trp 1,3,D-p-Cl-Phe 2,D-Phe 6, D-Ala 10]-LH-RH, most importantly contain new modification to positions 1, 2 and 10, and induce full blockade of ovulation at single doses as low as 10 μg per rat (50 μg/kg). Various ring substituents on D-Trp or D-Phe in position 1 or other D-amino acid replacements in position 10 did not significantly improve anti-ovulatory activity. Incorporation of N-Me-Leu in position 7 was slightly detrimental to activity

Detection of plasmin based on specific peptide substrate using acoustic transducer

© 2015 Elsevier B.V. All rights reserved. In this work we report the detection of plasmin protease by means of the thickness shear mode (TSM) acoustic method. The biorecognition element consists of a peptide substrate (PS) specific to plasmin immobilized on a piezoelectric quartz crystal electrode. After enzymatic reaction with plasmin, it cleaves a short fragment of the peptide causing increase in the resonance frequency of the piezo crystal. Plasmin was detected in the range of concentrations 1-20 nM, a target interval in which its presence presumably affects the quality of milk. The PS exhibited negligible response against to similar protease trypsin. This has been confirmed also by electrochemical detection method. Limit of detection of this acoustic transducer was found to be 0.65 nM. Formation of the sensing surface and kinetic effect of plasmin on the peptide substrate was studied by atomic force microscopy (AFM). The PS response was also validated in pretreated milk samples spiked by known concentrations of plasmin achieving an average recovery of 63 ± 0.6%

Detection of plasmin based on specific peptide substrate using acoustic transducer

© 2015 Elsevier B.V. All rights reserved. In this work we report the detection of plasmin protease by means of the thickness shear mode (TSM) acoustic method. The biorecognition element consists of a peptide substrate (PS) specific to plasmin immobilized on a piezoelectric quartz crystal electrode. After enzymatic reaction with plasmin, it cleaves a short fragment of the peptide causing increase in the resonance frequency of the piezo crystal. Plasmin was detected in the range of concentrations 1-20 nM, a target interval in which its presence presumably affects the quality of milk. The PS exhibited negligible response against to similar protease trypsin. This has been confirmed also by electrochemical detection method. Limit of detection of this acoustic transducer was found to be 0.65 nM. Formation of the sensing surface and kinetic effect of plasmin on the peptide substrate was studied by atomic force microscopy (AFM). The PS response was also validated in pretreated milk samples spiked by known concentrations of plasmin achieving an average recovery of 63 ± 0.6%