Genomic surveillance based on sequencing the entire genetic code of SARS-CoV-2 involves monitorin... more Genomic surveillance based on sequencing the entire genetic code of SARS-CoV-2 involves monitoring and studying genetic changes and variations in disease-causing organisms such as viruses and bacteria. By tracing the virus, it is possible to prevent epidemic spread in the community, ensuring a ‘precision public health’ strategy. A peptide-based design was applied to provide an efficacious strategy that is able to counteract any emerging viral variant of concern dynamically and promptly to affect the outcomes of a pandemic at an early stage while waiting for the production of the anti-variant-specific vaccine, which require longer times. The inhibition of the interaction between the receptor-binding domain (RBD) of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and one of the cellular receptors (DPP4) that its receptors routinely bind to infect human cells is an intriguing therapeutic approach to prevent the virus from entering human cells. Among the other modalities developed for this purpose, peptides surely offer unique advantages, including ease of synthesis, serum stability, low immunogenicity and toxicity, and small production and distribution chain costs. Here, we obtained a potent new inhibitor based on the rearrangement of a previously identified peptide that has been rationally designed on a cell dipeptidyl peptidase 4 (DPP4) sequence, a ubiquitous membrane protein known to bind the RBD-SPIKE domain of the virus. This novel peptide (named DPP4-derived), conceived as an endogenous “drug”, is capable of targeting the latest tested variants with a high affinity, reducing the VSV* DG-Fluc pseudovirus Omicron’s infection capacity by up to 14%, as revealed by in vitro testing in human Calu-3 cells. Surface plasmon resonance (SPR) confirmed the binding affinity of the new DPP4-derived peptide with Omicron variant RBD.
The use of GH treatment in subjects with a GH deficiency has led to contrasting results concernin... more The use of GH treatment in subjects with a GH deficiency has led to contrasting results concerning its impact to develop thyroid hyperfunction, whereas many others have underlined the possible onset of hypothyroidism. A number of studies have been carried out over short periods in subjects with multiple tropin deficiencies, in healthy adults or adults with GH deficiencies, in healthy adults or adults with GH deficiencies. The aim of the present study was to assess the effect of prolonged treatment with biosynthetic GH on thyroid function in children with an isolated idiopathic GH deficiency. The study included 8 children (mean age 10.4 +/- 0.8 years) with GH deficiencies treated with biosynthetic GH and 8 children with familial retarded stature of a similar age (mean age 10.3 +/- 0.7 years) who represented the control group. Serum levels of T3, T4, FT3, FT4 and TSH were measured at the start of the study and after one year of continuous GH treatment in subjects with GH deficiency; t...
The cellular uptake and the inhibitory effect of c-myb unmodified antisense oligonucleotides reve... more The cellular uptake and the inhibitory effect of c-myb unmodified antisense oligonucleotides reversibly bound to new polymeric nanoparticles in HL-60 cellular system have been found to increase by 50 folds if compared with the free ODN. An initial single dose (320 nM) of the nanoparticle bound unmodified antimyb ODN has been able to specifically inhibit HL-60 leukemia cell proliferation for at least 8 days.
The immunochemical study of the two hemoglobin components from the Arcid mollusc Scapharca inaequ... more The immunochemical study of the two hemoglobin components from the Arcid mollusc Scapharca inaequivalvis has been approached as a means to probe the quaternary structure of Arcid hemoglobins on the basis of the following considerations. Sequence changes on the surface of the molecule exceeding 40% usually suffice to eliminate immunological crossreactivity between genetically homologous proteins (1). In Arcid hemoglobins the three polypeptide chains that constitute the dimeric HbI and the tetrameric HbII display a characteristic pattern of variable and invariant regions; in the latter, notably the E and F helices, the homology reaches 75% and 100%, respectively (2–4). Thus, the presence or absence of crossreactivity between the two hemoglobins can be related in a simple way to the location of the homologous regions on the surface or in the interior of the molecule. The lack of crossreactivity between S. inaequivalvis HbI and HbII indicated that, in contrast to expectations based on the assembly of vertebrate hemoglobins, the E and F helices are not exposed to solvent (5) as shown by the low resolution X ray data of Royer et al. (6, and this volume)
Bovine spleen proteic inhibitors of serine proteases, belonging to the bovine pancreatic trypsin ... more Bovine spleen proteic inhibitors of serine proteases, belonging to the bovine pancreatic trypsin inhibitor (BPTI or aprotinin) family, have been localized, using immunocytochemical techniques, in the smooth muscle cells of some bovine spleen blood vessels. This vascular localization also occurs in a variety of bovine organs and differs from that of BPTI itself which is found exclusively in bovine mast cells, in agreement with previous reports. These data would be in favour of a possible involvement of one or more BPTI-type inhibitors in vascular processes by acting at the level of the smooth muscle cells, the tissue responsible for vasodilation/vasoconstriction events.
ABSTRACT The synthetic conjugate of the genotoxic compound 2,4 diaminotoluene (2,4 DAT) with gela... more ABSTRACT The synthetic conjugate of the genotoxic compound 2,4 diaminotoluene (2,4 DAT) with gelatin (2,4 DAT-GEL) was employed to elicit specific antibodies directed against a restricted class of aromatic diamines. Using this immunogen, mouse monoclonal antibodies (MAbs) have been produced. These MAbs have been characterized and used in ELISA to detect 2,4 DAT covalently linked to biopolymers. The MAbs could bind to different synthetic 2,4 DAT-biopolymer adducts as well as to DNA from rats treated in vivo with the aromatic diamine, but they did not react with gelatin or biopolymers alone. The use of these MAbs has been investigated in order to develop a highly sensitive test to detect adducts of this genotoxic compound with nuclear DNA.
A liposomal carrier system able to interact specifically with HL60 leukaemia cells was produced u... more A liposomal carrier system able to interact specifically with HL60 leukaemia cells was produced using small unilamellar liposomes made of pure phospholipids chemically cross-linked to human transferrin. The conjugation of transferrin to liposomes was carried out using N-succinimidyl 3-(2-pyridyldithio)-propionate and 2-iminothiolane as activating agents for the liposomes and the protein. The reaction occurred under conditions set to covalently link on the surface of a single vesicle a limited number (one to ten) of transferrin molecules, as verified by means of electron microscopy and immunoenzymic measurements. Before conjugation, the ultrastructure of the liposomes, and the content and distribution of the amino groups within the bilayer, were determined. The reactivity of the liposomes towards amino-derivatizing or thiolating compounds was also measured. Kinetic spectroscopic measurements confirmed that the distribution of the phosphatidylethanolamine in the vesicle bilayer is asy...
The effect of Lonidamine (LND), an energolytic chemosensitizing agent, on the MDR (multidrug resi... more The effect of Lonidamine (LND), an energolytic chemosensitizing agent, on the MDR (multidrug resistant) phenotype of a human breast cancer cell line (MCF-7) has been studied. The intracellular adriamycin (ADR) accumulation and distribution, the plasma membrane potential and the P170 glycoprotein phosphorylation, have been analysed after LND treatment. The analysis of the subcellular localisation of ADR in both wild type and resistant MCF-7 cells treated with ADR or ADR + LND revealed that LND induced an ADR intracellular redistribution in both cell lines. MCF-7 ADR resistant cells exposed to LND (50 micrograms/ml) showed a change in the electrical charges distribution across the plasma membrane and a time-dependent reduction of P170 phosphorylation (70% at 24 hr). These effects were associated with a marked increase in intracellular ADR accumulation in resistant cells.
Electrochemotherapy (ECT) is a new therapeutical technique that combines the administration of tr... more Electrochemotherapy (ECT) is a new therapeutical technique that combines the administration of trains of biphasic pulses with the local application of poorly permeant anticancer molecules, thus obtaining increased chemotherapy uptake. The purpose of this study was to prospectively assess the adjuvant potentialities of ECT for the treatment of different incompletely excised canine sarcomas. Twenty-two privately owned dogs with incomplete surgical excision of high grade sarcomas were treated with bleomycin injected within the tumor bed (1.5 IU/mg) followed by the sequential application of trains of biphasic pulses (8 pulses, 1300 V/cm, 50+50 micros duration, 1 Hz frequency). The overall response rate was 95% (21 out of 22 patients) with a mean time to recurrence of 730 days. At the time of writing 11 dogs were still in remission, three dogs had died of unrelated causes, one had local recurrence and the owner declined further treatment, one had limb amputation following recurrence, fou...
Genomic surveillance based on sequencing the entire genetic code of SARS-CoV-2 involves monitorin... more Genomic surveillance based on sequencing the entire genetic code of SARS-CoV-2 involves monitoring and studying genetic changes and variations in disease-causing organisms such as viruses and bacteria. By tracing the virus, it is possible to prevent epidemic spread in the community, ensuring a ‘precision public health’ strategy. A peptide-based design was applied to provide an efficacious strategy that is able to counteract any emerging viral variant of concern dynamically and promptly to affect the outcomes of a pandemic at an early stage while waiting for the production of the anti-variant-specific vaccine, which require longer times. The inhibition of the interaction between the receptor-binding domain (RBD) of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and one of the cellular receptors (DPP4) that its receptors routinely bind to infect human cells is an intriguing therapeutic approach to prevent the virus from entering human cells. Among the other modalities developed for this purpose, peptides surely offer unique advantages, including ease of synthesis, serum stability, low immunogenicity and toxicity, and small production and distribution chain costs. Here, we obtained a potent new inhibitor based on the rearrangement of a previously identified peptide that has been rationally designed on a cell dipeptidyl peptidase 4 (DPP4) sequence, a ubiquitous membrane protein known to bind the RBD-SPIKE domain of the virus. This novel peptide (named DPP4-derived), conceived as an endogenous “drug”, is capable of targeting the latest tested variants with a high affinity, reducing the VSV* DG-Fluc pseudovirus Omicron’s infection capacity by up to 14%, as revealed by in vitro testing in human Calu-3 cells. Surface plasmon resonance (SPR) confirmed the binding affinity of the new DPP4-derived peptide with Omicron variant RBD.
The use of GH treatment in subjects with a GH deficiency has led to contrasting results concernin... more The use of GH treatment in subjects with a GH deficiency has led to contrasting results concerning its impact to develop thyroid hyperfunction, whereas many others have underlined the possible onset of hypothyroidism. A number of studies have been carried out over short periods in subjects with multiple tropin deficiencies, in healthy adults or adults with GH deficiencies, in healthy adults or adults with GH deficiencies. The aim of the present study was to assess the effect of prolonged treatment with biosynthetic GH on thyroid function in children with an isolated idiopathic GH deficiency. The study included 8 children (mean age 10.4 +/- 0.8 years) with GH deficiencies treated with biosynthetic GH and 8 children with familial retarded stature of a similar age (mean age 10.3 +/- 0.7 years) who represented the control group. Serum levels of T3, T4, FT3, FT4 and TSH were measured at the start of the study and after one year of continuous GH treatment in subjects with GH deficiency; t...
The cellular uptake and the inhibitory effect of c-myb unmodified antisense oligonucleotides reve... more The cellular uptake and the inhibitory effect of c-myb unmodified antisense oligonucleotides reversibly bound to new polymeric nanoparticles in HL-60 cellular system have been found to increase by 50 folds if compared with the free ODN. An initial single dose (320 nM) of the nanoparticle bound unmodified antimyb ODN has been able to specifically inhibit HL-60 leukemia cell proliferation for at least 8 days.
The immunochemical study of the two hemoglobin components from the Arcid mollusc Scapharca inaequ... more The immunochemical study of the two hemoglobin components from the Arcid mollusc Scapharca inaequivalvis has been approached as a means to probe the quaternary structure of Arcid hemoglobins on the basis of the following considerations. Sequence changes on the surface of the molecule exceeding 40% usually suffice to eliminate immunological crossreactivity between genetically homologous proteins (1). In Arcid hemoglobins the three polypeptide chains that constitute the dimeric HbI and the tetrameric HbII display a characteristic pattern of variable and invariant regions; in the latter, notably the E and F helices, the homology reaches 75% and 100%, respectively (2–4). Thus, the presence or absence of crossreactivity between the two hemoglobins can be related in a simple way to the location of the homologous regions on the surface or in the interior of the molecule. The lack of crossreactivity between S. inaequivalvis HbI and HbII indicated that, in contrast to expectations based on the assembly of vertebrate hemoglobins, the E and F helices are not exposed to solvent (5) as shown by the low resolution X ray data of Royer et al. (6, and this volume)
Bovine spleen proteic inhibitors of serine proteases, belonging to the bovine pancreatic trypsin ... more Bovine spleen proteic inhibitors of serine proteases, belonging to the bovine pancreatic trypsin inhibitor (BPTI or aprotinin) family, have been localized, using immunocytochemical techniques, in the smooth muscle cells of some bovine spleen blood vessels. This vascular localization also occurs in a variety of bovine organs and differs from that of BPTI itself which is found exclusively in bovine mast cells, in agreement with previous reports. These data would be in favour of a possible involvement of one or more BPTI-type inhibitors in vascular processes by acting at the level of the smooth muscle cells, the tissue responsible for vasodilation/vasoconstriction events.
ABSTRACT The synthetic conjugate of the genotoxic compound 2,4 diaminotoluene (2,4 DAT) with gela... more ABSTRACT The synthetic conjugate of the genotoxic compound 2,4 diaminotoluene (2,4 DAT) with gelatin (2,4 DAT-GEL) was employed to elicit specific antibodies directed against a restricted class of aromatic diamines. Using this immunogen, mouse monoclonal antibodies (MAbs) have been produced. These MAbs have been characterized and used in ELISA to detect 2,4 DAT covalently linked to biopolymers. The MAbs could bind to different synthetic 2,4 DAT-biopolymer adducts as well as to DNA from rats treated in vivo with the aromatic diamine, but they did not react with gelatin or biopolymers alone. The use of these MAbs has been investigated in order to develop a highly sensitive test to detect adducts of this genotoxic compound with nuclear DNA.
A liposomal carrier system able to interact specifically with HL60 leukaemia cells was produced u... more A liposomal carrier system able to interact specifically with HL60 leukaemia cells was produced using small unilamellar liposomes made of pure phospholipids chemically cross-linked to human transferrin. The conjugation of transferrin to liposomes was carried out using N-succinimidyl 3-(2-pyridyldithio)-propionate and 2-iminothiolane as activating agents for the liposomes and the protein. The reaction occurred under conditions set to covalently link on the surface of a single vesicle a limited number (one to ten) of transferrin molecules, as verified by means of electron microscopy and immunoenzymic measurements. Before conjugation, the ultrastructure of the liposomes, and the content and distribution of the amino groups within the bilayer, were determined. The reactivity of the liposomes towards amino-derivatizing or thiolating compounds was also measured. Kinetic spectroscopic measurements confirmed that the distribution of the phosphatidylethanolamine in the vesicle bilayer is asy...
The effect of Lonidamine (LND), an energolytic chemosensitizing agent, on the MDR (multidrug resi... more The effect of Lonidamine (LND), an energolytic chemosensitizing agent, on the MDR (multidrug resistant) phenotype of a human breast cancer cell line (MCF-7) has been studied. The intracellular adriamycin (ADR) accumulation and distribution, the plasma membrane potential and the P170 glycoprotein phosphorylation, have been analysed after LND treatment. The analysis of the subcellular localisation of ADR in both wild type and resistant MCF-7 cells treated with ADR or ADR + LND revealed that LND induced an ADR intracellular redistribution in both cell lines. MCF-7 ADR resistant cells exposed to LND (50 micrograms/ml) showed a change in the electrical charges distribution across the plasma membrane and a time-dependent reduction of P170 phosphorylation (70% at 24 hr). These effects were associated with a marked increase in intracellular ADR accumulation in resistant cells.
Electrochemotherapy (ECT) is a new therapeutical technique that combines the administration of tr... more Electrochemotherapy (ECT) is a new therapeutical technique that combines the administration of trains of biphasic pulses with the local application of poorly permeant anticancer molecules, thus obtaining increased chemotherapy uptake. The purpose of this study was to prospectively assess the adjuvant potentialities of ECT for the treatment of different incompletely excised canine sarcomas. Twenty-two privately owned dogs with incomplete surgical excision of high grade sarcomas were treated with bleomycin injected within the tumor bed (1.5 IU/mg) followed by the sequential application of trains of biphasic pulses (8 pulses, 1300 V/cm, 50+50 micros duration, 1 Hz frequency). The overall response rate was 95% (21 out of 22 patients) with a mean time to recurrence of 730 days. At the time of writing 11 dogs were still in remission, three dogs had died of unrelated causes, one had local recurrence and the owner declined further treatment, one had limb amputation following recurrence, fou...
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Papers by G. Citro
ensuring a ‘precision public health’ strategy. A peptide-based design was applied to provide an efficacious strategy that is able to counteract any emerging viral variant of concern dynamically and promptly to affect the outcomes of a pandemic at an early stage while waiting for the production
of the anti-variant-specific vaccine, which require longer times. The inhibition of the interaction between the receptor-binding domain (RBD) of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and one of the cellular receptors (DPP4) that its receptors routinely bind to infect
human cells is an intriguing therapeutic approach to prevent the virus from entering human cells. Among the other modalities developed for this purpose, peptides surely offer unique advantages, including ease of synthesis, serum stability, low immunogenicity and toxicity, and small production and distribution chain costs. Here, we obtained a potent new inhibitor based on the rearrangement of a previously identified peptide that has been rationally designed on a cell dipeptidyl peptidase 4
(DPP4) sequence, a ubiquitous membrane protein known to bind the RBD-SPIKE domain of the virus. This novel peptide (named DPP4-derived), conceived as an endogenous “drug”, is capable of targeting the latest tested variants with a high affinity, reducing the VSV* DG-Fluc pseudovirus
Omicron’s infection capacity by up to 14%, as revealed by in vitro testing in human Calu-3 cells. Surface plasmon resonance (SPR) confirmed the binding affinity of the new DPP4-derived peptide with Omicron variant RBD.
ensuring a ‘precision public health’ strategy. A peptide-based design was applied to provide an efficacious strategy that is able to counteract any emerging viral variant of concern dynamically and promptly to affect the outcomes of a pandemic at an early stage while waiting for the production
of the anti-variant-specific vaccine, which require longer times. The inhibition of the interaction between the receptor-binding domain (RBD) of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and one of the cellular receptors (DPP4) that its receptors routinely bind to infect
human cells is an intriguing therapeutic approach to prevent the virus from entering human cells. Among the other modalities developed for this purpose, peptides surely offer unique advantages, including ease of synthesis, serum stability, low immunogenicity and toxicity, and small production and distribution chain costs. Here, we obtained a potent new inhibitor based on the rearrangement of a previously identified peptide that has been rationally designed on a cell dipeptidyl peptidase 4
(DPP4) sequence, a ubiquitous membrane protein known to bind the RBD-SPIKE domain of the virus. This novel peptide (named DPP4-derived), conceived as an endogenous “drug”, is capable of targeting the latest tested variants with a high affinity, reducing the VSV* DG-Fluc pseudovirus
Omicron’s infection capacity by up to 14%, as revealed by in vitro testing in human Calu-3 cells. Surface plasmon resonance (SPR) confirmed the binding affinity of the new DPP4-derived peptide with Omicron variant RBD.