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J de la Fuente

Publications and source records attributed to J de la Fuente.

At least 19 recordsLinked to original sources
PubMed2007

Development and pattern of mRNA relative abundance of bovine embryos cultured in the isolated mouse oviduct in organ culture.

The aim of this study was to examine the development of bovine zygotes in isolated mouse oviducts (IMO) and the quality of the blastocysts produced. In vitro produced bovine zygotes were transferred into the ampullae of the IMO and cultured in SOF or KSOM. Control embryos were cultured in droplets of the same media. Following 6 days of culture, blastocysts were processed for nuclei counts or mRNA abundance. Culture in the IMO did not affect the proportion of zygotes developing to the blastocyst stage compared to the respective control droplets (SOF: 17.7 +/- 3.2% vs. 18.8 +/- 2.7%; KSOM: 20.7 +/- 2.6% vs. 22.2 +/- 2.8%). Culture in the IMO in KSOM resulted in an increased number of inner cell mass (ICM) nuclei; however, total nuclei number or incidence of apoptosis was unaffected. Culture in the IMO in SOF resulted in an increase (P < 0.05) in abundance of transcripts in blastocysts for Oct-4 and SOX, and reduced abundance of Glut-1, Na/K, Cx43, and survivin compared to blastocysts derived from culture in SOF alone. In contrast, culture in the IMO in KSOM resulted in increased abundance of transcripts for Glut-1, Cx43, Oct-4, and survivin and reduced expression of Na/K and SOX compared to KSOM alone. Transcripts for G6PDH, IFN-tau, and E-Cad were unaffected. These data confirm that the IMO is capable of supporting development of bovine embryos. Depending on the basal medium used, the pattern of transcript abundance in embryos derived from the IMO is similar to that of in vivo derived embryos.

Animals↗
PubMed2007

Experimental infection of C3H/HeJ mice with the NY18 isolate of Anaplasma phagocytophilum.

Human granulocytic anaplasmosis (HGA), an emerging disease of public health concern in many areas of the world, is caused by Anaplasma phagocytophilum. Small animal models of A phagocytophilum in laboratory mice have been developed and used to study the pathogenesis of HGA. In this study, we characterized the pathologic changes in acute infection of C3H/HeJ mice experimentally infected with the NY18 isolate of A phagocytophilum. Although no clinical signs were noted, acute infection was associated with gross splenomegaly, microscopic inflammatory lesions in the lung and liver, hyperplastic lesions on the spleen, and clinical pathology abnormalities including neutropenia and monocytosis. This study emphasizes the use of well-defined animal models as a valuable tool for the study of A phagocytophilum infections.

Anaplasma phagocytophilum↗
PubMed2006

Renal transplantation in diabetic patients.

OBJECTIVE: Diabetes is one of the main causes of end-stage renal disease (ESRD) and admission to hemodialysis, and the demand for kidney transplantation in this population has increased. Our aim was to evaluate the clinical aspects and survival of diabetic patients with kidney transplants by comparing them with the nondiabetic population. MATERIALS AND METHODS: Patients transplanted during the period from 1994 to 2003 were evaluated for this study. The transplant and demographic characteristics were analyzed by the chi-square test and Student t test according to the type of variable. Kaplan-Meier curves and the log-rank test were used to evaluate the graft and patient survival. RESULTS: From a total of 523 consecutive renal transplants, 35 (6.6%) were diabetics who were older than nondiabetics (47 +/- 11 years vs 37 +/- 16, P < .002). Patients received immunosuppression with cyclosporine (84.3%), tacrolimus (11.2%), azathioprine (46.6%), mycophenolate mofetil (43.5%), and steroids (all patients). The diabetic patients had a higher percentage of living donors (33.5% vs 17.2%; P = .04). Graft survival rates at 1, 3, and 5 years were 82.7%, 70.9%, and 63.0% in the diabetic patients and 87.6%, 79.0%, and 72.5% (P = .6) in the nondiabetic patients. Patient survival at 5 years was 90.5% in diabetic patients vs 89.0% in nondiabetic patients (P = .9). CONCLUSIONS: No differences were found in our series in transplant complications or survival in the diabetic patients compared with the nondiabetic patients. Kidney transplants, even with living donors, must be offered to well-selected diabetic patients without reservations.

Diabetes Mellitus, Type 1↗
PubMed2006

Strategies for development of vaccines for control of ixodid tick species.

Ticks are distributed worldwide and impact human and animal health, as well as food animal production. Control of ticks has been primarily by application of acaricides, which has resulted in selection of resistant ticks and environmental pollution. Vaccines have been shown to be a feasible tick control method that offers a cost-effective, environmentally friendly alternative to chemical control. However, identification of tick-protective antigens remains the limiting step in vaccine development. Tick antigens exposed naturally to the host during tick feeding and those concealed have both shown promise as candidate vaccine antigens. Development of vaccines against multiple tick species may be possible using highly conserved tick-protective antigens or by antigens showing immune cross-reaction to different tick species. Vaccines made from a combination of key protective antigens may greatly enhance vaccine efficacy. Preliminary studies have suggested the possibility of vaccine strategies directed toward both tick control and the blocking of pathogen transmission. Characterization of the tick genomes will have a great impact on the discovery of new protective antigens. The future of research directed toward tick vaccine development is exciting because of new and emerging technologies for gene discovery, and vaccine formulation and delivery.

Animals↗
PubMed2005

Infection with Anaplasma phagocytophilum in a seronegative patient in Sicily, Italy: case report.

BACKGROUND: Anaplasma phagocytophilum causes human granulocytic anaplasmosis (HGA) in humans, which has been recognized as an emerging tick-borne disease in the United States and Europe. Although about 65 cases of HGA have been reported in Europe, some of them do not fulfill the criteria for confirmed HGA. Confirmation of HGA requires A. phagocytophilum isolation from blood, and/or identification of morulae in granulocytes and/or positive PCR results with subsequent sequencing of the amplicons to demonstrate specific rickettsial DNA. Seroconversion or at least fourfold increase in antibody titers to A. phagocytophilum has been used as criteria for confirmed HGA also. CASE PRESENTATION: Infection with A. phagocytophilum was confirmed by PCR in a patient in Sicily, Italy, who had negative serology for A. phagocytophilum. A fragment of A. phagocytophilum 16S rDNA was amplified by two independent laboratories and sequenced from two separate patient's blood samples. The 16S rDNA sequence was identical in both samples and identical to the sequence of the A. phagocytophilum strain USG3 originally obtained from a dog. CONCLUSION: Infection with A. phagocytophilum was confirmed in a patient without a detectable antibody response against the pathogen. The results reported herein documented the first case of confirmed HGA in Sicily, Italy. These results suggested the possibility of human infections with A. phagocytophilum strains that result in clinical symptoms and laboratory findings confirmatory of HGA but without detectable antibodies against the pathogen.

Anaplasma phagocytophilum↗
PubMed2005

Genetic diversity of Anaplasma marginale strains from cattle farms in the province of Palermo, Sicily.

Bovine anaplasmosis, caused by the tick-borne rickettsia Anaplasma marginale, is endemic in Sicily and results in economic loss to the cattle industry. This study was designed to characterize strains of A. marginale at the molecular level from cattle in the Province of Palermo, Sicily. Seropositivity of cattle >or=1 year old for A. marginale in the study area ranged from 62% to 100%. The observed prevalence of A. marginale infections in cattle herds ranged from 25% to 100%. Two predominant A. marginale msp4 genotypes were found. A positive correlation was found between the prevalence of infection and the presence of Rhipicephalus (Boophilus) annulatus. Phylogenetic analysis of msp4 sequences of European strains of A. marginale did not provide phylogeographical information. These results suggest that development of farm husbandry systems and vaccines for genetically heterogeneous populations of A. marginale are needed for control of anaplasmosis in this region of Sicily.

Anaplasma marginale↗
PubMed2005

Serologic cross-reactivity between Anaplasma marginale and Anaplasma phagocytophilum.

In the context of a serosurvey conducted on the Anaplasma marginale prevalence in Swiss cattle, we suspected that a serological cross-reactivity between A. marginale and A. phagocytophilum might exist. In the present study we demonstrate that cattle, sheep and horses experimentally infected with A. phagocytophilum not only develop antibodies to A. phagocytophilum (detected by immunofluorescent-antibody assay) but also to A. marginale (detected by a competitive enzyme-linked immunosorbent assay). Conversely, calves experimentally infected with A. marginale also developed antibodies to A. phagocytophilum using the same serological tests. The identity of 63% determined in silico within a 209-amino-acid sequence of major surface protein 5 of an isolate of A. marginale and one of A. phagocytophilum supported the observed immunological cross-reactivity. These observations have important consequences for the serotesting of both, A. marginale and A. phagocytophilum infection of several animal species. In view of these new findings, tests that have been considered specific for either infection must be interpreted carefully.

Anaplasma marginale↗
PubMed2005

[Sterile maggots as adjuvant procedure for local treatment in a patient with proximal calciphylaxis].

Proximal calciphylaxis with skin ulcerations has a very poor survival and infection is the main cause of the high mortality rate. We present the case of a diabetic obese hemodialysis woman diagnosed of severe calciphylaxis with extensive ulcers in the abdomen and left thigh. After a first ineffective debridement attempt, systemic medical treatment was associated with daily wet cures with enzymatic ointments and maggot therapy resulting in spectacular granulation and healing of these ulcers. We want to underline the great properties of sterile maggots to selectively dissolve necrotic tissue, to disinfect the wound and to stimulate healing. Thus, sterile-maggot debridement may constitute a very good adjuvant treatment even in proximal ulcers of calciphylaxis, preventing systemic infection.

Animals↗
PubMed2004

The risks of translocating wildlife. Pathogenic infection with Theileria sp. and Elaeophora elaphi in an imported red deer.

It is well known that the translocation of wild animals poses risks of the introduction of pathogens into populations, and regulations and recommendations regarding quarantine and screening protocols for wild animals do exist. Less is known about the infection of imported animals with local endemic pathogens. A red deer stag that had been imported from Germany was found recumbent and died from hemolytic anaemia and a process of exertional myopathy. Infection with Theileria sp. was detected in thin blood smears and confirmed by PCR and sequencing. In addition, massive parasitation by Elaeophora elaphi, a parasite endemic to Iberian red deer, was detected. Sequence comparison between the 18S rRNA gene sequence determined that the Theileria strain involved in this case had a 99.7% identity with a Theileria sp. strain obtained from sika-deer, and 95.3% identity with T. cervi. Using sequence distance analysis, the strain from red deer grouped with isolates from Cervus spp. as opposed to isolates from Odocoileus spp. and bovines. Both detected parasites are of little pathogenicity to local red deer, but were pathogenic for the imported red deer from Northern Europe. This case demonstrates that local endemic pathogens may pose naive translocated animals at risk, and illustrates the need for thorough examination and planification of translocation protocols.

Animals↗
PubMed2004

Effect of speed of development on mRNA expression pattern in early bovine embryos cultured in vivo or in vitro.

Recent data have demonstrated that fast-cleaving embryos produced in vitro are more likely to develop to blastocyst stage, and that the postfertilization culture system used impacts considerably on the mRNA expression and quality of blastocysts produced. The present study is the first to investigate the relationship between the developmental speed of embryos produced in vivo or in vitro and the temporal transcription pattern. Genes related to important preimplantation events are monitored during the first 4 days of embryo development in embryos with fast or slow development. The set of genes analyzed in the present study characterizes several important physiological processes including: transport and metabolism of fructose (Glut-5), stress (SOX), mitochondrial activity and detoxification of reactive oxygen species (MnSOD), cell communication (Cx43), maternal recognition of pregnancy (IFN-tau), imprinting (IGF-II), apoptosis (Bax), growth factor binding and metabolism (IGF-IR), and oxidative stress (G6PD). Using real time PCR, we have found that for all the genes analyzed there are differences in mRNA expression between embryos with fast and slow developmental speed produced both in vitro and in vivo. Frequently, genes that may be stress induced such as SOX, MnSOD, BAX, IFtau, and G6PD were highly transcribed in in vitro produced embryos and in embryos with slow developmental speed. On the other side, transcripts from genes related with metabolism, growth, and differentiation (Glut-5, Cx 43, IGF-II, and IGF-IR) were detected in higher amounts in in vivo produced embryos and in embryos with fast developmental speed. Moreover, it is interesting to stand out that for some genetic markers (such as SOX and G6PD) there are in vivo and in vitro differences that can be observed even before materno-zygotic transition, which probably reflects a differential mRNA degradation. These transcription patterns reflects the embryonic response to the adverse in vitro culture conditions, and connect the low quality of embryos which slow developmental speed produced in vivo and in vitro, with the mRNA expression pattern of some embryonic genes.

Animals↗
PubMed2004

Anaplasma marginale (Rickettsiales: Anaplasmataceae): recent advances in defining host-pathogen adaptations of a tick-borne rickettsia.

The tick-borne intracellular pathogen Anaplasma marginale (Rickettsiales: Anaplasmataceae) develops persistent infections in cattle and tick hosts. While erythrocytes appear to be the only site of infection in cattle, A. marginale undergoes a complex developmental cycle in ticks and transmission occurs via salivary glands during feeding. Many geographic isolates occur that vary in genotype, antigenic composition, morphology and infectivity for ticks. In this chapter we review recent research on the host-vector-pathogen interactions of A. marginale. Major surface proteins (MSPs) play a crucial role in the interaction of A. marginale with host cells. The MSP1a protein, which is an adhesin for bovine erythrocytes and tick cells, is differentially regulated and affects infection and transmission of A. marginale by Dermacentor spp. ticks. MSP2 undergoes antigenic variation and selection in cattle and ticks, and contributes to the maintenance of persistent infections. Phylogenetic studies of A. marginale geographic isolates using msp4 and msp1alpha provide information about the biogeography and evolution of A. marginale: msp1alpha genotypes evolve under positive selection pressure. Isolates of A. marginale are maintained by independent transmission events and a mechanism of infection exclusion in cattle and ticks allows for only the infection of one isolate per animal. Prospects for development of control strategies by use of pathogen and tick-derived antigens are discussed. The A. marginale/vector/host studies described herein could serve as a model for research on other tick-borne rickettsiae.

Anaplasma marginale↗
PubMed2004

Assessing relationships between health-related quality of life and adherence to antiretroviral therapy.

OBJECTIVE: To investigate associations between health-related quality of life (HRQoL), as assessed using the multidimensional quality of life-HIV (MQOL-HIV) questionnaire, and adherence to antiretroviral treatment in HIV-infected subjects. DESIGN: Multicentre cross-sectional study in three institutional tertiary hospitals in northwest Spain. PATIENTS AND METHODS: The MQOL-HIV was completed by 235 HIV-infected adults undergoing antiretroviral treatment. Adherence to antiretroviral therapy was assessed by using patient's self-report. Information about sociodemographic characteristics and clinical variables was also collected. RESULTS: Good adherence (> or = 95% of prescribed pills correctly taken) was reported by 131 patients (55.7%). Univariate analyses indicated that the sociodemographic and clinical variables associated with adherence were age, educational level, income, employment, home stability, transmission route, history of previous antiretroviral therapy, and number of prescribed pills/day. Subscales of MQOL-HIV associated with adherence were mental health, cognitive functioning, financial status, medical care, partner intimacy, and (in men only) sexual functioning. Stepwise logistic regression showed that good adherence was more frequent in patients aged > 40 years (odds ratio, OR: 2.50; 95% confidence interval, CI: 1.15-5.61) and in patients with high cognitive functioning (OR: 2.26; 95% CI: 1.19-4.30). Conversely, poor adherence was more frequent in patients without stable home (OR: 2.96; 95% CI: 1.39-6.32), in patients required to take 14 or more pills/day (OR: 2.17; 95% CI: 1.18-4.28), in patients with low financial status (OR: 3.42; 95% CI: 1.57-7.45), and in patients reporting low medical care (OR: 2.07; 95% CI: 1.07-3.98). CONCLUSIONS: HRQoL dimensions, notably cognitive functioning, financial status and medical care, are closely associated with antiretroviral therapy adherence.

Adult↗
PubMed2003

Temporal divergence in the pattern of messenger RNA expression in bovine embryos cultured from the zygote to blastocyst stage in vitro or in vivo.

The objective of this study was to examine the time during the postfertilization period that gene expression patterns in in vitro-cultured bovine embryos diverge from those of their in vivo-cultured counterparts. Presumptive bovine zygotes were produced by in vitro maturation and fertilization of immature oocytes collected from the ovaries of slaughtered animals. Approximately 20 h post insemination (hpi), zygotes were denuded and randomly divided into two groups for culture either in vitro, in synthetic oviduct fluid medium, or in vivo, in the ewe oviduct. Embryos were recovered from both systems at approximately 30 hpi (2-cell), 2 (4-cell), 3 (8-cell), 4 (16-cell), 5 (early morula), 6 (compact morula), or 7 (blastocyst) days post insemination. On recovery, they were examined for stage of development and snap frozen in liquid nitrogen for the analysis of transcript abundance using real-time polymerase chain reaction. The transcripts studied were glucose transporter 5, sarcosine oxidase, mitochondrial Mn-superoxide dismutase, connexin 43, interferon tau, insulin-like growth factor II, apoptosis regulator box-alpha and insulin-like growth factor-I receptor, most of which are known from our previous work to differ in terms of transcript abundance in blastocysts derived from culture in vitro or in vivo. The results demonstrate that the relative abundance of the transcripts studied varies throughout the preimplantation period and is strongly influenced by the culture environment. In addition, the data demonstrate that changes in transcript abundance in blastocyst stage embryos are in many cases a consequence of perturbed transcription earlier in development. Depending on the transcript, these differences may be evident by as little as 10 h of initiation of culture. Such information has implications not only for basic biology but also for human assisted reproduction in which there is a move toward culturing embryos to the blastocyst stage, necessitating prolonged culture in vitro under potentially deleterious conditions.

Animals↗
PubMed2003

Non-TBI stem cell transplantation protocol for Fanconi anaemia using HLA-compatible sibling and unrelated donors.

Allogeneic haemopoietic stem cell transplantation (SCT) is the only curative option for severe bone marrow (BM) failure in patients with Fanconi anaemia (FA). We have developed a non total body irradiation (TBI) conditioning protocol consisting of fludarabine (120-150 mg/m(2)), low dose of cyclophosphamide (40 mg/kg) and antilymphocyte globulin (45 mg/kg). Graft-versus-host disease (GVHD) prophylaxis was with cyclosporin alone for sibling allografts but also included Campath-1 H (days 1-5 post SCT) for the unrelated allografts. We have performed two sibling and two unrelated BM transplants with a follow-up of 11-51 months. All patients experienced minimal toxicity and were discharged from hospital 28-32 days post SCT. Neutrophil and platelet engraftment occurred from days 11 to 19 and 15 to 34, respectively. All patients achieved stable full donor haemopoiesis with normalisation of the peripheral blood count despite one of them having myelodysplasia (MDS) with 8% blasts prior to the SCT. The only site of acute GVHD was in the skin (grade I-II) and only one patient progressed to limited chronic GVHD. This protocol is associated with reduced toxicity and prompt engraftment in FA patients with AA and/or MDS undergoing SCT using sibling or unrelated donors.

Adolescent↗
PubMed2003

Identification of fetal mesenchymal stem cells in maternal blood: implications for non-invasive prenatal diagnosis.

Strategies for genetic prenatal diagnosis on fetal cells in the maternal circulation have been limited by lack of a cell type present only in fetal blood. However, the recent identification of mesenchymal stem cells (MSC) in first trimester fetal blood offers the prospect of targeting MSC for non-invasive prenatal diagnosis. We developed protocols for fetal MSC enrichment from maternal blood and determined sensitivity and specificity in mixing experiments of male fetal MSC added to female blood, in dilutions from 1 in 10(5) to 10(8). We then used the optimal protocol to isolate fetal MSC from maternal blood in the first trimester, using blood taken after surgical termination of pregnancy as a model of increased feto-maternal haemorrhage. In model mixtures, we could amplify one male fetal MSC in 2.5 x 10(7) adult female nucleated cells, yielding a 100% pure population of fetal cells, but not one fetal MSC in 10(8) nucleated cells. Fetal MSC were identified in one of 20 post-termination maternal blood samples and confirmed as fetal MSC by XY fluorescence in-situ hybridization (FISH), immunophenotyping and osteogenic and adipogenic differentiation. We report the isolation of fetal MSC from maternal blood; however, their rarity in post-termination blood suggests they are unlikely to have a role in non-invasive prenatal diagnosis. Failure to locate these cells routinely may be attributed to their low frequency in maternal blood, to sensitivity limitations of enrichment technology, and/or to their engraftment in maternal tissues soon after transplacental passage. We speculate that gender microchimerism in post-reproductive maternal tissues might result from feto-maternal trafficking of MSC in early pregnancy.

Adult↗