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Biomedical subjects

G Ricca

Publications and source records attributed to G Ricca.

At least 19 recordsLinked to original sources
PubMed2004

Polyalkylphenyl-sulphonic acids with acid groups of variable strength from animal-vegetable wastes.

The insoluble organic fraction isolated from rice-hulls residues and animal fecal matter mixture is sulphonated in liquid SO3 at 200 degrees C to a water soluble sulphonate (III). III is compared to the sulphonate (IV) obtained from the same mix after composting. Both products have been found to contain mixtures of molecules with close molecular weight. These molecules consist of a central cicloaliphatic cluster with peripheral pending aromatic chains. III and IV appear to have the same sulphonation degree. However, the latter contains higher concentrations of cicloaliphatic fragments and of amide, phenol and ether bonds, but less carboxylic and amine functional groups. These differences may be reasonably traced back to the starting materials. By comparison with commercial lignosulphonates derived from the paper and pulp industry, the above arylsulphonates are likely candidates for a variety of applications in the chemical industry and in agriculture. We conclude that sulphonation, even under the drastic experimental conditions of this work, does not seem to erase the memory of the parent matter structure. This reaction is capable of upgrading recalcitrant organic matter in vegetable waste residues to an interesting variety of lignosulphonates.

Agriculture↗
PubMed2003

Polyalkylphenyl-sulphonic acid with acid groups of variable strength from compost.

The insoluble organic fraction (humin-like material, HLM) from rice hull-dairy cattle compost is well converted into water soluble HLM-sulphonate by reaction in liquid SO3. Microanalytical, potentiometric, molecular weight, and NMR data are consistent with a highly homogeneous polymeric arylsulphonate having 4000 Da MW, 1.3 sulphonic groups per aromatic ring and significant content of carboxylic and phenolic groups. By comparison with structure-property relationships for commercial lignosulphonates derived from the pulp and paper industry, the above arylsulphonate is likely to be a candidate at a variety of applications in the chemical industry and in agriculture. Therefore, sulphonation is a means for upgrading composts HLM to the same uses as for commercial lignosulphonates.

Agriculture↗
PubMed2003

Swallowing disorders: management data.

Aim of the investigation was to assess the workload and verify the results of oropharyngeal dysphagia management in a large state hospital by means of a descriptive, observational prospective study and descriptive statistical analysis. 81 patients [37 females, 44 males, mean age 61.3 (+/- 13) years] suffering from oropharyngeal dysphagia were evaluated and treated in the in- and outpatient Divisions of the "Azienda Ospedaliera S. Giovanni Battista" in Turin. Treatment of oropharyngeal dysphagia included changes in consistency and texture of food, compensatory postures of head, strengthening exercises for oropharyngeal muscles, and stimulation of pharyngeal sensitivity. In data collection and analysis, the following were used as outcome measures: mode of nutrition delivery (oral, enteral, parenteral), dietary adjustments, presence of aspiration or penetration, and use of compensatory head positioning. Results showed that the number of patients fed by parenteral or enteral tube (50/81 prior to treatment) dropped to 36/81 upon discharge from hospital. Those unable to take anything by mouth, from 55 dropped to 9. The number of patients with aspiration or penetration dropped, respectively, from 47 and 8 to 20 and 4. Postural changes were used in 15 cases. Data obtained indicate that oropharyngeal dysphagia rehabilitation outcomes are promising. Better understanding of the rheological characteristics of food and a stricter, more rigorous evaluation of the outcomes on activities and social participation are warranted.

Aged↗
PubMed1996

Disposition of a novel recombinant antithrombotic agent, RG 12986, in cynomolgus monkeys.

RG 12986, a novel antagonist of platelet aggregation, is a recombinant peptide based on the sequence in von Willebrand factor, which contains the GP1b binding site. Disposition of the peptide in cynomolgus monkeys was determined using nonlabeled and 35S-labeled product. After iv administration, the peptide underwent a triphasic decay in the plasma. The first phase of elimination, after distribution, had a t1/2 (approximately 20 min) similar to that observed for inhibition of platelet aggregation (approximately 25 min). The correlation between the logarithm of the plasma peptide concentration and activity was r = 0.9989. The effective duration of pharmacological activity was approximately 2 hr. After this period, a slower terminal phase of plasma elimination was observed (t1/2 approximately 2 hr). Plasma clearance (7-15 ml/min/kg) and volume of distribution at steady-state (0.4-0.9 L/kg) estimates appeared to have a slight dose dependency, but the scope of the investigation did not allow this to be verified. There was a linear correlation between dose and AUC (r2 = 0.9998), but for each 4-fold increase in dose there was a greater than 4-fold increase in AUC. Immediately after iv administration, significant fragmentation of the peptide was observed with polyacrylamide gel electrophoresis analysis of the plasma. This initial rate of metabolism was subsequently slowed to t1/2 estimates of 2 hr, followed by a very long terminal half-life of plasma radioactivity of 12 days. It is likely that this terminal half-life represents metabolic recycling of 35S. Elimination of the label was primarily via the kidneys.

Animals↗
PubMed1992

Expression of human serum albumin in the milk of transgenic mice.

We have tested the feasibility of producing large quantities of human serum albumin (HSA) in the milk of transgenic livestock by generating transgenic mice as a model system. The sheep beta-lactoglobulin (BLG) 5'-regulatory promoter sequences were used to support expression of BLG or HSA in transgenic mice. Transgenic animals generated from the entire BLG gene including 3, 5.5 or 10.8 kb of 5'-sequences demonstrated that 3 kb of 5'-sequences were sufficient to support high levels of expression of BLG, and that the longer 5'-sequences did not improve upon the levels of expression. As such, the 3 kb 5'-sequences were used to drive expression of HSA in BLG-HSA constructs. HSA was not detectably expressed in eight transgenic lines generated from a BLG-HSA construct containing the HSA cDNA. Two transgenic lines of 26 generated, using five different constructs, with an HSA minigene possessing the first intron expressed HSA in their milk. One of these expressed HSA at high levels (2.5 mg ml-1) and has stably transmitted this ability to its progeny. A high percentage of transgenic mouse lines (four of six) generated from a vector containing an HSA minigene possessing introns 1 and 2 expressed HSA in their milk at levels which ranged from 1 to 35 micrograms ml-1. In a similar trend, levels of expression of HSA by transfected tissue culture cells from BLG-HSA vectors containing an introduced SV40 enhancer were low with the HSA cDNA, increased with the HSA minigene with intron 1 and increased further with the minigene containing introns 1 and 2. This study demonstrates that high levels of HSA can be expressed in the milk of transgenic animals, that introns of the HSA gene play a role in its expression and that transfected cell lines may be used to quickly evaluate the relative expression efficiencies of various vector constructs intended for future transgenic evaluation.

Animals↗
PubMed1992

Production and functional characterization of a recombinant fragment of von Willebrand factor (vWF): an antagonist to platelet receptor GP Ib.

We expressed a recombinant peptide fragment (Ser445-Val733) of human von Willebrand factor (vWF), containing the binding domain for the platelet receptor of GP Ib, in E. coli. This 33 kD peptide blocks binding of the intact vWF molecule to GP Ib in the presence of modulators. Thus, it offers potential as an antithrombotic agent. High level expression was achieved in a plasmid construct driven by the bacteriophage T7 promoter. The peptide was solubilized from inclusion bodies in strong chaotrope, then reduced and alkylated. Following purification, formulation at pH 3.5, and lyophilization, the reconstituted experimental product (RG 12986) exists as an equilibrium of monomer and dimer species. When formulated above pH 5.0, soluble aggregates are formed; these solutions have less bioactivity than RG 12986. Interestingly, the non-aggregated state of RG 12986 remains conserved following dilution and incubation with platelet-poor plasma. The overall purification/low pH formulation strategies may be applicable to other E. coli recombinant proteins having a tendency to aggregate following removal of chaotrope near physiologic pH when in a concentrated format.

Cloning, Molecular↗
PubMed1991

Functional modulation of the isolated glycoprotein Ib binding domain of von Willebrand factor expressed in Escherichia coli.

We have expressed in Escherichia coli the domain of von Willebrand factor (vWF) containing the binding site for platelet glycoprotein (GP) Ib and used it to study the regulation of vWF-platelet interaction. The recombinant fragment, comprising residues 445-733 of the mature vWF subunit and designated rvWF445-733, did not have the native conformation of the corresponding domain in the intact molecule because, in order to prevent formation of random aggregates, the seven cysteine residues in the sequence were reduced and alkylated. Unlike native vWF, rvWF445-733 bound to GP Ib in the absence of any modulator, suggesting that the lack of disulfide bonds and/or carbohydrate side chains within this domain may expose platelet interaction sites. In the presence of two modulators, the glycopeptide ristocetin and the snake protein botrocetin, rvWF445-733 inhibited native vWF binding to GP Ib as well as platelet aggregation mediated by vWF, suggesting that both the fragment and the native molecule interact with the same site on platelets. This conclusion was also supported by the observation that the recombinant fragment competed with the binding to platelets of an anti-GP Ib monoclonal antibody known to inhibit vWF binding. Botrocetin formed a complex with rvWF445-733, but the affinity of this interaction was approximately 25-fold lower than with native vWF. However, the complexes of botrocetin with either rvWF445-733 or multimeric native vWF bound to GP Ib with similar dissociation constant. Therefore, conformational attributes of vWF regulate its affinity for botrocetin, but once the complex is formed, interaction with GP Ib is independent of native vWF conformation. These findings provide insights into the regulation of vWF-platelet interaction.

Binding Sites↗
PubMed1990

Cloning of three human tyrosine phosphatases reveals a multigene family of receptor-linked protein-tyrosine-phosphatases expressed in brain.

A human brainstem cDNA library in bacteriophage lambda gt11 was screened under conditions of reduced hybridization stringency with a leukocyte common antigen (LCA) probe that spanned both conserved cytoplasmic domains. cDNA encoding a receptor-linked protein-tyrosine-phosphatase (protein-tyrosine-phosphate phosphohydrolase, EC 3.1.3.48), RPTPase alpha, has been cloned and sequenced. Human RPTPase alpha consists of 802 amino acids. The extracellular domain of 150 residues includes a hydrophobic signal peptide and eight potential N-glycosylation sites. This is followed by a transmembrane region and two tandemly repeated conserved domains characteristic of all RPTPases identified thus far. The gene for RPTPase alpha has been localized to human chromosome region 20pter-20q12 by analysis of its segregation pattern in rodent-human somatic cell hybrids. Northern blot analysis revealed the presence of two major transcripts of 4.3 and 6.3 kilobases. In addition to RPTPase alpha, two other RPTPases (beta and gamma), identified in the same screen, have been partially cloned and sequenced. Analysis of sequence comparisons among LCA, the LCA-related protein LAR, and RPTPases alpha, beta, and gamma reveals the existence of a multigene family encoding different RPTPases, each containing a distinct extracellular domain, a single hydrophobic transmembrane region, and two tandemly repeated conserved cytoplasmic domains.

Amino Acid Sequence↗
PubMed1990

[Modification of the blink reflex in hemidystonia].

With the aim of evaluating the excitability of the brain stem reflex centers, we studied the side-to-side differences in the EMG activity of the early and late components of the blink reflex, in subjects with unilateral dystonia without demonstrable brain lesions. We observed that both early and late responses of direct blink reflex were significantly higher in the affected side than in the contralateral one.

Blinking↗
PubMed1990

Unilateral enhancement of early and late blink reflex components in hemidystonia.

The side-to-side differences in the EMG activity of the early and late components of blink reflex, regarded as revealing the state of excitability of the brain stem reflex centers, have been analyzed in patients with unilateral dystonia without demonstrable brain lesions. It has been observed that both early and late responses of direct blink reflex were higher on the side affected by hemidystonia than on the contralateral one, while the latency values were in the normal range. Possibility that an abnormal output from the striatum towards the brainstem structures involved in blink reflex appears on the affected side of hemidystonic patients is discussed.

Adult↗
PubMed1988

Influences of dopaminergic systems on the blink reflex.

The effect of the anti-dopaminergic drug haloperidol (6 mg/day i.m. for 2 days) on the blink reflex elicited by electric stimulation of supraorbital nerves was investigated in 12 adult volunteers. A significant increase in amplitude of the early component (R1) of the reflex was observed. This increase disappeared within 5 days of stopping the drug. Possible neural systems which might be involved in these effects are considered.

Adult↗
PubMed1987

Molecular cloning of the human gene for von Willebrand factor and identification of the transcription initiation site.

A series of overlapping cosmid genomic clones have been isolated that contain the entire coding unit of the human gene for von Willebrand factor (vWf), a major component of the hemostatic system. The cloned segments span approximately 175 kilobases of human DNA sequence, and hybridization analysis suggests that the vWf coding unit is approximately 150 kilobases in length. Within one of these clones, the vWf transcription initiation site has been mapped and a portion of the vWf promoter region has been sequenced, revealing a typical "TATA box," a downstream "CCAAT box," and a perfect downstream repeat of the 8 base pairs containing the transcription start site. Sequencing of a segment of another genomic clone has revealed the vWf translation termination codon. Where tested, comparative restriction analysis of cloned and chromosomal DNA segments strongly suggests that no major alterations occurred during cloning and that there is only one complete copy of the vWf gene in the human haploid genome. Similar analyses of DNA from vWf-producing endothelial cells and nonexpressing leukocytes suggest that vWf gene expression is not accompanied by gross genomic rearrangements. In addition, there is significant homology of C-terminal coding sequences among the vWf genes of several vertebrate species.

Amino Acid Sequence↗
PubMed1986

Isolation and sequencing of a cDNA clone homologous to the v-sis oncogene from human endothelial cells.

A clone containing the 3' end of the mRNA for the human c-sis gene (homologous to the B chain of platelet-derived growth factor) was isolated from a cDNA library derived from human umbilical vein endothelial cells and then sequenced. The analysis of possible translation products in all three reading frames indicated that the A chain of platelet-derived growth factor was not coded for within the 3' end of the c-sis mRNA. The 3' end of the mRNA for c-sis is contained in or adjacent to exon 6.

Base Sequence↗