Biomedical subjects

J M Alonso

Publications and source records attributed to J M Alonso.

At least 91 records · Page 5Linked to original sources

PubMed1995

Immunodetection of the large form of the gamma 2 subunit of mammalian GABAA receptor.

Two forms of the GABAA receptor gamma 2-subunit, named short (gamma 2S) and long (gamma 2L), and generated by alternative RNA splicing, have been identified in mammalian brain by molecular cloning techniques. We have produced antibodies against a synthetic peptide containing the 8-amino acid insertion present in the long form but not in the short one. Using the antipeptide serum, we have identified the gamma 2L subunit in membrane preparations of GABAA receptors from rat and mouse cerebellum and its cellular location in cerebellum.

Alternative Splicing↗

PubMed1995

Order-disorder in T, T', and T* phase: superconductors and related materials.

After reviewing microstructural studies on superconducting materials showing T, T', and T* structural types, results are presented on the microstructure of some n-type superconductors and related materials prepared with accurate control of the oxygen stoichiometry. Electron microscopy is used to describe the ordering of interstitial oxygen defects in T-type La2NiO4 + delta leading to the formation of the n = 2 term of a homologous series with the general formula La8nNi4nO16n + 1. Structural transitions and superstructure formation in the Pr2-x-yCexSryCuO4-delta system are reported, where T, T', and T* phases are isolated as a function of both Ce and Sr content.

Electric Conductivity↗

PubMed1995

A non-photosynthetic ferredoxin gene is induced by ethylene in Citrus organs.

The sequence and expression of mRNA homologous to a cDNA encoding a non-photosynthetic ferredoxin (Fd1) from Citrus fruit was investigated. The non-photosynthetic nature of this ferredoxin was deduced from: (1) amino acid sequence alignments showing better scores with non-photosynthetic than with photosynthetic ferredoxins, (2) higher expression in tissues containing plastids other than chloroplast such as petals, young fruits, roots and peel of fully coloured fruits, and (3) the absence of light-dark regulation characteristic of photosynthetic ferredoxins. In a phylogenetic tree constructed with higher-plant ferredoxins, Citrus fruit ferredoxin clustered together with root ferredoxins and separated from the photosynthetic ferredoxins. Non photosynthetic (root and fruit) ferredoxins, but not the photosynthetic ferredoxins, have their closest homologs in cyanobacteria. Analysis of ferredoxin genomic organization suggested that non-photosynthetic ferredoxins exist in Citrus as a small gene family. Expression of Fd1 is developmentally regulated during flower opening and fruit maturation, both processes may be mediated by ethylene in Citrus. Exogenous ethylene application also induced the expression of Fd1 both in flavedo and leaves. The induction on non-photosynthetic ferredoxins could be related with the demand for reducing power in non-green, but biosynthetically active, tissues.

Amino Acid Sequence↗

PubMed1995

Evidence for the involvement of ethylene in the expression of specific RNAs during maturation of the orange, a non-climacteric fruit.

Twelve cDNAs corresponding to mRNAs inducible by ethylene were isolated by differential screening of a cDNA library from ethylene-treated Citrus sinensis fruits. Northern analysis of RNA extracted from flavedo of ethylene-treated fruits and from fruits at different maturation stages showed that some of the mRNAs corresponding to these cDNAs were regulated both by ethylene treatment and during fruit maturation. The effect of exogenous ethylene on leaves and of endogenous ethylene on flowers showed that gene induction was not restricted to the flavedo tissue. The possible role of ethylene during maturation of the non-climacteric Citrus fruit is discussed.

Blotting, Northern↗

PubMed1995

A putative vacuolar processing protease is regulated by ethylene and also during fruit ripening in Citrus fruit.

A putative citrus vacuolar processing thiolprotease cDNA (Cit-vac) was isolated from a cDNA library of Citrus fruits (Citrus sinensis L. Osbeck var Washington navel). The cDNA is 58 and 57% identical with vacuolar processing seed proteases from castor bean and soybean, respectively. The Citvac sequence shows a typical signal peptide for entering into the endoplasmic reticulum and two glycosylation signals. Using an in vitro transcription-translation system, we show that the Citvac precursor is able to enter a microsomal fraction and to undergo proteolytic processing and glycosylation. Transcript levels for the Citvac are developmentally regulated in the flavedo (outer colored part of the fruit peel) and increase during fruit ripening and in the flower during opening. Exogenous treatment with ethylene induces Citvac mRNA expression in both fruits and leaves. Citvac is encoded by one or two genes in the Citrus genome. The possible role of the Citvac gene product during fruit ripening and other ethylene-mediated processes is discussed.

Amino Acid Sequence↗

PubMed1994

An indirect fluorescent antibody technique for detection of anti-Dermatophilus congolensis antibodies in sheep.

An indirect fluorescent antibody (IFA) technique has been developed for detection of anti-Dermatophilus antibodies in sheep. Sera from 25 bacteriologically confirmed clinically affected sheep and from 10 negative non affected lambs were used. Whole cell antigen from brain heart infusion cultures of D. congolensis was used and all sera were tested in the same way for cross-reactivity against antigens obtained from cultures of Actinomyces viscosus, Micrococcus luteus, Nocardia asteroides, and Corynebacterium pseudotuberculosis. Sera from Dermatophilus-infected sheep gave positive results with D. congolensis antigen and negative results with the antigens from other bacteria. The whole cell antigens employed were simple to prepare and easy to recognise by microscopy. Cross-reactivity was further tested using the D. congolensis culture whole cell antigen and 3 sera from sheep with bacteriologically confirmed natural infections due to Corynebacterium pseudotuberculosis, Actinomyces pyogenes and Nocardia asteroides. None of these sera showed positive reactions. The authors recommend this technique for serological surveys and research on dermatophilosis.

Actinomycetales↗

PubMed1994

Modulatory influence of putative inhibitors of nitric oxide synthesis on visual processing in the cat lateral geniculate nucleus.

1. Using an in vivo preparation we have examined the actions of two inhibitors of nitric oxide synthase (NOS), NG-nitro-L-arginine (L-NOArg) and NG-methyl-L-arginine (L-MeArg), in the feline dorsal lateral geniculate nucleus (dLGN). We compared the responses obtained to iontophoretic application of these substances during visual stimulation with those elicited by visual stimulation alone. The effects of concurrent ejection of L-arginine (L-Arg), the normal physiological substrate of NOS, and D-arginine, the inactive isomer, were tested on these responses. 2. Extracellular application of L-NOArg and L-MeArg produced clear and repeatable effects, consisting of substantial reduction in discharge rate without affecting response selectivity, on 94% of tested cells. These effects were prevented by simultaneous application of L-Arg, which when ejected alone produced no change on visual evoked responses. 3. The data suggest that nitric oxide (NO) is necessary for the transmission of the visual input under normal visual stimulation and show a direct involvement of NO in visual information processing at the level of dLGN, suggesting that its contribution to brain mechanisms is more profound than previously thought.

Amino Acid Oxidoreductases↗

PubMed1994

[Familial thrombophilia due to resistance to activated protein C].

Resistance to activated protein C (RAPC) has been described recently as a cause of trombophilia; this may justify up to 50% of thromboembolic disease without predisposing cause in patients under 45 years. A 29 years-old male with a previous deep venous thrombosis (DVT) in the lower left limb three years earlier, developed a DVT in the right lower limb after a trauma of the knee that required immobilization, was associated to pulmonary thromboembolism diagnosed by gammagraphic methods. The phlebographic study showed femoro-iliaco-caval venous thrombosis. The proband's father and a younger brother had a previous history of thrombotic episodes. The following tests, were performed in the proband and relatives: prothrombin time, aPTT, thrombin time, fibrinogen, (Von Clauss), antithrombin III (chromogenic), protein C and protein S (coagulometry and ELISA), plasminogen (chromogenic) and lupus anticoagulant (ITT, dRVVT, aCL). RAPC was evaluated in two different samples. The proband study was performed under oral anticoagulation treatment (OAT). Control groups were: 21 blood donors and 12 OAT patients. The results showed a decreased response to APC in the proband (ratio 1.5) and relatives: father (1.4), brothers (1.5 and 1.5), while the mother was within the normal range (> or = 2). In normal controls and OAT patients the ratio was over 2. No other abnormalities were detected in the assays performed. It is concluded that RAPC is the cause of this familial trombophilia. RAPC should be included in the evaluation study of trombophilia.

Adolescent↗

PubMed1993

Cell responses to vertical and horizontal retinal disparities in the monkey visual cortex.

Because of the horizontal separation of both ocular globes, the projection angles are slightly different. These differences are commonly termed retinal disparities. Vertical and horizontal retinal disparities occur constantly in normal life. We have investigated the responses of single cells in cortical areas V1 and V2 of behaving Macaca mulatta monkeys to retinal disparities by using dynamic random dot stereograms. Our findings show that cortical visual cells are sensitive to both vertical and horizontal disparities. To calculate the distance between two objects in a three-dimensional space from horizontal disparities, it is necessary to know the fixation distance. It has been suggested that the horizontal gradient of vertical disparity contains information to estimate the fixation distance and therefore to scale horizontal disparities. We suggest that these cells sensitive to horizontal and vertical disparities represent a neural mechanism that provides information to the visual system in order to achieve a correct eye alignment and depth perception.

Animals↗

PubMed1993

Effect of bromoconduritol on glucosidase II from rat liver. A new kinetic model for the binding and hydrolysis of the substrate.

Bromoconduritol inhibits the p-nitrophenyl-glucosidase and maltase activities of glucosidase II purified from rat liver, an enzyme that removes the two alpha-1,3-linked glucose residues of the protein-bound oligosaccharide Glc2Man9GlcNAc2 in the processing of N-glycoproteins. The inactivation process exhibits pseudo-first-order kinetics. Previously, we have demonstrated the occurrence of two binding (active) sites in the glucosidase II for the substrates p-nitrophenyl alpha-D-glucopyranoside (pNphGlc) and maltose (high- and low-affinity sites). The inhibition kinetic studies with bromoconduritol indicate that the high- and low-affinity sites for pNphGlc correspond to high- and low-affinity sites for maltose, respectively. Bromoconduritol has no effect on the binding of the substrates (pNphGlc and maltose) to the high-affinity site, although it does modify the low-affinity site and hinders the binding of the two indicated substrates to this site. These results, together with previous reports, have prompted us to propose a new kinetic model of binding and hydrolysis of the physiological substrate of the enzyme, in which the outermost glucose residue would bind and be released at the high-affinity site, whereas the innermost glucose residue would do so at the low-affinity site.

Animals↗

PubMed1993

Functional double-negative T cells in the periphery express T cell receptor V beta gene products that cause deletion of single-positive T cells.

A proportion of peripheral T cells lack surface expression of the CD4 or CD8 coreceptor molecules and hence are designated as " double negative" (DN). Most DN T lymphocytes express the gamma/delta T cell receptor (TcR), but a minor fraction of them, in both humans and mice, express the alpha/beta TcR. Whereas alpha/beta+ DN T lymphocytes are infrequent (< 1%) in conventional lymphoid organs (spleen, blood, lymph node), they account for two-thirds of the T cells residing in adult bone marrow. Analysis of the TcR V beta repertoire expressed by peripheral DN T cells revealed a high frequency of cells bearing autoreactive TcR that cause deletion of "single-positive" (SP) (CD4+CD8-or CD4-CD8+) T cells. Peripheral DN cells thus represent a cell type that is relatively resistant to clonal deletion. Furthermore, such cells have not been inactivated (anergized) in vivo since they proliferate and secrete interleukins in response to cross-linking by monoclonal antibodies specific for these V beta gene products that are deleted in SPT cells. These results might help to understand the association of peripheral expansion of DN cells and development of autoimmune diseases.

Animals↗

PubMed1993

Orientational influences of layer V of visual area 18 upon cells in layer V of area 17 in the cat cortex.

We examined the orientation tuning curves of 86 cells located in layer V of area 17, before, during, and after focal blockade of a small (300-microns diameter) region of near-retinotopic register in layer V of area 18 of quantitatively established orientation preference. Such focal blockade revealed three distinct populations of area 17 layer V cells-cells with decreased responses to stimuli of some orientations (21%), cells with increased responses to stimuli of some orientations (43%), and cells unaffected by the focal blockade (36%). These effects were clearcut, reproducible, and generally directly related to the known receptive field properties of the cell recorded in area 18 at the center of the zone of blockade. These effects were also analyzed in terms of alterations in orientation bandwidth in the cells in area 17 as a result of the blockade-bandwidth increases (22%) and decreases (24%) were found; however, these changes were essentially unrelated to the measured receptive field properties. Inhibitory and excitatory effects were most pronounced when the regions in areas 17 and 18 were of like ocular dominance and were of similar orientation preference. Inhibitory effects (suggesting a normally excitatory input) were most dependent upon the similarity of receptive fields; excitatory effects (suggesting a normally inhibitory input) were less heavily dependent.

Animals↗

PubMed1993

Influence of layer V of area 18 of the cat visual cortex on responses of cells in layer V of area 17 to stimuli of high velocity.

Focal blockade of restricted regions in layer V of area 18 was used to assess the contribution of this region to the responses to high-velocity stimuli of cells in retinotopically matched, layer V in area 17. In 40% of cases, blockade within area 18 revealed responses of area 17 cells to high-velocity stimuli to which they previously showed only poor responses. Stimulus specificity of the cells in area 17 was otherwise unaltered. All effects were reversible and repeatable. We suggest that a component of the output of layer V from area 18 normally suppresses the responses of retinotopically matched cells within area 17 to stimuli of high velocity, thereby enhancing the specificity of those cells to stimuli of low velocity.

Animals↗

PubMed1993

Binocular matching in monkey visual cortex: single cell responses to correlated and uncorrelated dynamic random dot stereograms.

This paper presents results obtained from experiments performed on two behaving monkeys (Macaca mulatta) viewing correlated and uncorrelated dynamic random dot stereograms. Extracellular single unit activity was recorded from areas V1 and V2. We found that cells sensitive to correlated stereograms (38%) were also sensitive to uncorrelated stereograms. Regarding their response to both situations, these cells were grouped into two types. The first type (57%), termed "correlation-sensitive cells", responded with statistically significant difference to correlated and uncorrelated stereograms. The second type responded equally to both stereograms (43%), and therefore they were termed "correlation-insensitive cells". Our data suggest that these two types may form two populations with different functions subserving depth perception.

Animals↗

PubMed1993

Enzymatic activities of Dermatophilus congolensis measured by API ZYM.

API ZYM kit was used to test enzymatic activities on eighteen strains of Dermatophilus congolensis. All strains produced lipase and acid phosphatase, which act on lipids, and leucine arylamidase which act on proteins. Another 10 exoenzymes were present in at least one of the strains.

Actinomycetales↗

PubMed1993

Geographical variations in the prevalence of HIV infection among drug users receiving ambulatory treatment in Spain.

To identify the factors responsible for the regional differences in HIV-positive serostatus among drug users in Spain receiving outpatient treatment, the 17 autonomous regions into which Spain is divided were classified as high (mean 52%) or low prevalence groups (mean 34%) depending on the prevalence of seropositivity. In regions where the prevalence of positive serostatus was high, unemployment was more markedly associated with HIV infection than in low prevalence regions, while other potential risk factors yielded the same strength of association. Even so, adjustment for all the factors only accounted for 13% of the total difference in HIV+ prevalence between regions. The current distribution of risk factors among the two groups of autonomous regions does not explain these differences.

Acquired Immunodeficiency Syndrome↗

PubMed1993

Passive local immunotherapy of experimental staphylococcal pneumonia with human intravenous immunoglobulin.

Staphylococcus aureus remains a life-threatening agent of nosocomial pneumonia in immunocompromised patients. The increasing incidence of strains exhibiting wide-spectrum resistance to antibiotics, such as methicillin-resistant S. aureus (MRSA), requires new therapeutic strategies. There is renewed interest in passive immunization with human plasma-derived immunoglobulins (IVIG) as antiinfective agents. The efficacy of IVIG was tested in an experimental model of staphylococcal pneumonia, using both an MRSA clinical isolate and reference strain Cowan III, in mice immunosuppressed with cyclophosphamide. Efficient antistaphylococcal activities were obtained with IVIG administered intravenously or intranasally. IVIG saturated with protein A or its F(ab')2 fragments were as efficient as intact IVIG, suggesting that protection did not require opsonization through IgG Fc-phagocyte Fc gamma-receptor interactions. Thus, topical administration of IVIG may replace a local antibody response to S. aureus in an immunocompromised host and may be useful in prophylaxis and treatment of nosocomial S. aureus pneumonia.

Animals↗

PubMed1993

Characterization of the maltase activity of glucosidase II from rat liver. Kinetic model.

Glucosidase II is a key enzyme in the processing of N-glycoproteins since it removes the two glucose residues from the protein-linked oligosaccharide Glc2Man9GlcNAc2-R. We have studied the kinetics of the purified enzyme, using maltose as substrate. Analysis of data fitting to single and double-hyperbolic equations and the Eadie-Hofstee profile indicate that the enzyme has two binding (active) sites for the hydrolysis of maltose. The Km and Vmax values for the high-affinity site were 0.43 mM and 691 mU/mg, respectively, whereas the values for the low-affinity site were 57.7 mM and 2888 mU/mg, respectively. The Vmax/Km ratios were 1607 and 50.1 ml/min per g for the high- and low-affinity sites, respectively. A new kinetic model for this enzyme is proposed from the equilibria corresponding to the partial competitive inhibition produced by maltose on p-nitrophenyl-glucosidase activity. The amino acid composition of the enzyme has been established.

Amino Acids↗