Biomedical subjects

C Carrillo

Publications and source records attributed to C Carrillo.

At least 19 recordsLinked to original sources

PubMed2004

Spatial effects in discrete generation population models.

A framework is developed for constructing a large class of discrete generation, continuous space models of evolving single species populations and finding their bifurcating patterned spatial distributions. Our models involve, in separate stages, the spatial redistribution (through movement laws) and local regulation of the population; and the fundamental properties of these events in a homogeneous environment are found. Emphasis is placed on the interaction of migrating individuals with the existing population through conspecific attraction (or repulsion), as well as on random dispersion. The nature of the competition of these two effects in a linearized scenario is clarified. The bifurcation of stationary spatially patterned population distributions is studied, with special attention given to the role played by that competition.

Animals↗

PubMed2004

Passive protection to bovine rotavirus (BRV) infection induced by a BRV VP8* produced in plants using a TMV-based vector.

We have previously reported on the use of a tobacco mosaic virus (TMV) vector TMV-30B to express foreign viral antigens for use as experimental immunogens. Here we describe the development of an improved TMV-30B vector that adds a sequence of 7 histidine residues to the C-terminus of recombinant proteins expressed in the vector. We used this TMV-30B-HISc vector to express the VP8* fragment of the VP4 protein from bovine rotavirus (BRV) strain C-486 in plants. Recombinant VP8* protein was purified from N. benthamiana leaves at 7 days post-inoculation by immobilized metal affinity chromatography. The plant-produced VP8* was initially detected using anti-His tag mAb and its antigenic nature was confirmed using both monoclonal and polyclonal specific antisera directed against BRV. Adult female mice, inoculated by the intraperinoteal route with an immunogen containing 4 microg of recombinant VP8*, developed a specific and sustained response to the native VP8* from the homologous BRV. Eighty five percent of suckling mice from immunized dams that were challenged with the homologous virus at the fifth day of age were protected from virus as compared to 35% of the pups from mothers immunized with a control protein. These results demonstrate that the plant-produced VP8* was able to induce passive protection in the new born through the immunization of dams. This suggests that the technology presented here provides a simple method for using plants as an inexpensive alternative source for production of recombinant anti-rotavirus antigens.

Animals↗

PubMed2002

Genes coding structural proteins in the Leishmania braziliensis complex.

Acidic ribosomal P1 and P2b proteins, referred to as P proteins, and histone H3 are reported for first time in the Leishmania braziliensis complex. Deoxyribonucleic acid analysis and multiple sequence alignment suggest that both P proteins may maintain their structural function in the ribosomal stalk, in spite of the high rate of mutations detected. The deduced amino acid sequence of protein P1 showed 51% identity with Trypanosoma cruzi protein P1 and protein P2b showed 61% identity with T. cruzi protein P2b. Another conserved protein, L. (Viannia) braziliensis histone H3, showed 82% and 70% identity with histone H3 of L. (Leishmania) infantum and T. cruzi, respectively. The N-terminal end of this histone is divergent in comparison with the consensus eukaryotic sequence. Their predicted tridimensional structure was designed.

Amino Acid Sequence↗

PubMed2001

Seroprevalence of bovine leukemia virus in dairy cattle in Argentina: comparison of sensitivity and specificity of different detection methods.

Bovine leukemia virus (BLV) is a retrovirus that induces a chronic infection in cattle, which develop in three possible pathological forms: asymptomatic course, persistent lymphocytosis (PL) and lymphosarcoma. Once infected, cattle remain virus carriers for life and start to show a serological reaction within a few weeks after infection. Eradication and control of the disease is based on early diagnostic and segregation of the carriers. The agar gel immunodiffusion (AGID) test has been the serological test of choice for routine diagnosis of serum samples. Nevertheless, in more recent years, the enzyme-linked immunosorbent assay (ELISA) has replaced the AGID for large scale testing. Although Argentina has over 60 million cattle population, no nationwide studies have been conducted yet to determine the prevalence of the infection. To estimate the rate of BLV infection in dairy cattle in Argentina, a survey for specific antibodies in >10,000 serum samples from animals over 18 months old, belonging to 363 different herds from the largest dairy production areas of the country, was carried out in our laboratory, along 1999. For this purpose, we developed an ELISA to detect serum antibodies against the BLV virus. The cut-off of the ELISA was established over 339 serum samples, using polymerase chain reaction and southern blot (PCR-SB) as confirmatory test. The sensitivity and specificity of the ELISA was of 97.2 and 97.5%, respectively, while the local official AGID test showed a sensitivity of 79.7% and specificity of 99.0%. To know the seroprevalence of BLV on dairy herds, and also the incidence of the infection within the herd, the serological survey was based on individual serum samples. The results show that the prevalence of infected individuals is of 32.85%, while the percentage of infected herds, harboring one or more infected animals, is of 84%. These results indicate a medium level of seropositive animals when taken individually, but a high prevalence of infected farms, which has been notoriously increased in the last 15 years as shown when compared with previous data from particular geographic areas, indicating that BLV constitutes a serious sanitary problem for dairy producers in Argentina. They also indicate the poor sensitivity of the official AGID test used in the country.

Animals↗

PubMed2001

Variation in sequence and RNA editing within core domains of mitochondrial group II introns among plants.

The 3' regions of several group II introns within the mitochondrial genes nad1 and nad7 show unexpected sequence divergence among flowering plants, and the core domains 5 and 6 are predicted to have weaker helical structure than those in self-splicing group II introns. To assess whether RNA editing improves helical stability by the conversion of A-C mispairs to A-U pairs, we sequenced RT-PCR amplification products derived from excised intron RNAs or partially spliced precursors. Only in some cases was editing observed to strengthen the predicted helices. Moreover, the editing status within nad1 intron 1 and nad7 intron 4 was seen to differ among plant species, so that homologous intron sequences shared lower similarity at the RNA level than at the DNA level. Plant-specific variation was also seen in the length of the linker joining domains 5 and 6 of nad7 intron 3; it ranged from 4 nt in wheat to 11 nt in soybean, in contrast to the 2-4 nt length typical of classical group II introns. However, this intron is excised as a lariat structure with a domain 6 branchpoint adenosine. Our observations suggest that the core structures and sequences of these plant mitochondrial introns are subject to less stringent evolutionary constraints than conventional group II introns.

Alternative Splicing↗

PubMed2001

Analysis of fatal pedestrian injuries in Mexico City, 1994-1997.

Currently, in Mexico City, 57% of deaths from traffic crashes are pedestrian injuries. The purpose of this study was to identify and analyse the magnitude, trends, risks, and geographic distribution of fatal pedestrian injuries during the period 1994-1997. A cross-sectional study design was used, based on the death certificates of persons who were in Mexico City during 1994-1997 and died due to pedestrian injuries. Mortality rates, trend analysis, standard mortality ratios (SMRs), and potential years of life lost index (PYLLI) were calculated by gender, age groups and region (these correspond to "delegations", which are political divisions of the city). Rates were derived according to place of occurrence and place of residence. A total of 3687 pedestrian fatalities were reported, and 71% of these were to Mexico City residents. The rate for males was 10.6/100000 with a slight decrease in rate from 1994 to 1997. For females, the rate was 4.0/100000, with an increase in rate during the same years. The present study allowed identification of the target population as well as regions with a high risk of fatal pedestrian injuries. From these data we have developed or recommended specific interventions for prevention and control of fatal pedestrian injuries in Mexico City.

Accidents, Traffic↗

PubMed2001

Induction of a virus-specific antibody response to foot and mouth disease virus using the structural protein VP1 expressed in transgenic potato plants.

We have recently communicated the oral and parental immunogenicity of the structural protein VP1 of foot and mouth disease virus (FMDV) expressed in different transgenic plants. Those results clearly indicated the necessity of increasing the expression of the foreign genes in the transgenic plant to avoid additional steps toward the purification and/or concentration of the antigen of interest. Here, we report the production of transgenic potatoes plants containing the VP1 gene cloned under the regulatory activity of either a single (pRok2) or a double (pRok3) copy of the S35 cauliflower mosaic virus (CaMV 35S) promoter, as a strategy for increasing the level of VP1 gene expression. The presence of the VP1 gene in the plants was confirmed by polymerase chain reaction (PCR) and its specific transcription activity was demonstrated by reverse transcriptase-polymerase chain reaction (RT-PCR). The results showed that, although the immunized animals presented a FMDV VP1 specific antibody response and protection against the experimental challenge, no significant differences were demonstrated in the immunizing activity of plant extracts obtained from the pRok2 or pRok3 transformed plants. These results confirm those previously obtained using other plant species allowing the possibility of using plants as antigen expression vectors, and demonstrated that at least in the potato system, the use of double CaMV 35S promoter does not cause a significant increase in the level of the VP1 expressed.

Animals↗

PubMed2000

Sensitivity of trypanosomatid protozoa to DFMO and metabolic turnover of ornithine decarboxylase.

alpha-Difluoromethylornithine (DFMO), the specific and irreversible inhibitor of ornithine decarboxylase (ODC), was able to induce the arrest of proliferation in Leishmania mexicana and ODC-transformed Trypanosoma cruzi cultures grown in a semi-defined medium essentially free of polyamines. Conversely, Crithidia fasciculata and Phytomonas 274 were not affected by the inhibitor. The drug-resistance of Crithidia and Phytomonas was neither caused by an impairment of DFMO uptake nor by a decrease of the enzyme affinity for the inhibitor. We were also able to rule out the possibility of ODC overexpression in the drug-tolerant parasites. The measurements of ODC metabolic turnover indicated that the enzymes from Crithidia and Phytomonas have a short half-life of 20-40 min, while ODC from Leishmania and transgenic Trypanosoma cruzi are rather stable with a half-life longer than 6 hours. Analyses of polyamine internal pools under different growth conditions have shown that DFMO was able to markedly decrease the levels of putrescine and spermidine in all parasites, but the depletion of spermidine was higher in trypanosomatids containing an ODC with slow turnover. Our results suggest that in these parasites cultivated in the presence of the drug, spermidine might decrease below critical levels needed to maintain trypanothione concentrations or other conditions essential for normal proliferation.

Animals↗

PubMed2000

Risk factors in highway traffic accidents: a case control study.

The objective of this study was to identify risk factors related to the driver, the vehicle and the environment, that are associated with motor vehicles accidents on highways. A case and control design was used where the case was: the driver of a motor vehicle who was involved in an accident while traveling on the Mexico-Cuernavaca highway; and the control was: the driver or a motor vehicle who, traveling on the highway, completed the trip without being involved in a traffic accident. Risk factors associated with the occurrence of a traffic accident (P > 0.05) were: age under 25 years, frequent travel, traveling to work, alcohol consumption, driving during the daylight, on a week-day, under adverse weather conditions and in the Mexico Cuernavaca direction of road. Risks adjusted by logistic regression were: age under 25 years odds ratio (OR) 3.01, confidence interval (CI) (95%) 1.46-6.18; work as a travel reason OR 1.74, CI 1.06 2.86; alcohol intake OR 4.70, Cl 1.62 13.6; driving under adverse weather conditions OR 5.70, CI 3.66-8.85; traveling on a week-day OR 1.84, CI 1.14-2.94; during daylight hours OR 4.23, CI 2.36-7.58 and in the Mexico Cuernavaca direction of road OR 2.69, CI 1.67 4.32. The identification of factors associated with the risk of being involved in a highway traffic accident allows us to propose primary prevention measures for this important public health problem.

Accidents, Traffic↗

PubMed1999

Sp1 and AP2 transcription factors are required for the human fragile mental retardation promoter activity in SK-N-SH neuronal cells.

The promoter of the human fragile mental retardation gene (FMR1) was functionally analyzed in order to identify elements responsible for its regulation. Plasmids carrying the wild type or different deleted-promoter sequences driving the chloramphenicol acetyl transferase gene (CAT) were transiently transfected into the SK-N-SH cells and the CAT activity was assessed. Deletion studies suggested that major regulatory elements are present in a DNA region between positions -123 and -51. Gel mobility shift and footprinting assays using a DNA fragment encompassing that promoter region showed that SP1 and AP2 transcription factors could be involved in the functioning of the FMR1 promoter.

Cell Line↗

PubMed1999

Protection of mice against challenge with foot and mouth disease virus (FMDV) by immunization with foliar extracts from plants infected with recombinant tobacco mosaic virus expressing the FMDV structural protein VP1.

A tobacco mosaic virus (TMV)-based vector has been used to express in plants the complete open reading frame coding for VP1, the major immunogenic protein of foot and mouth disease virus (FMDV). In vitro RNA transcripts were inoculated into Nicotiana benthamiana plants and detectable amounts of recombinant VP1 were identified by Western blot as soon as 4 days postinfection. Foliar extracts prepared from infected leaves were injected intraperitoneally into mice and all of the immunized animals developed a specific antibody response to both the complete virus particle and the major immunogenic region as determined by ELISA and Western blot analysis. Most importantly, all immunized mice developed a protective immune response against experimental challenge with virulent FMDV. To our knowledge, this is the first report showing the expression of a complete open reading frame of an antigenic foreign protein in plants, using a recombinant plant virus, in sufficient quantity to permit use of the crude plant extract as an experimental immunogen to protect animals against virus challenge.

Animals↗

PubMed1999

Trypanosoma cruzi epimastigotes lack ornithine decarboxylase but can express a foreign gene encoding this enzyme.

Trypanosoma cruzi, a pathogenic protozoan causing Chagas disease, lacks ornithine decarboxylase (ODC), the enzyme catalyzing the first step of polyamine biosynthetic pathway in eukaryotic cells. Our results indicate that the auxotrophy for diamines of T. cruzi epimastigotes is due to the absence of an active ODC gene in these parasites and not to the inability for the expression of this gene. The introduction of an exogenous complete coding region from Crithidia fasciculata ODC gene inserted in an expression vector specific for trypanosomatids induces the normal expression of the foreign genetic information allowing the transformed T. cruzi to overcome the exogenous polyamine requirement for growth. The enzyme expressed in the transformed parasites has shown a considerably extended metabolic stability. The loss of ODC activity in T. cruzi might be related to the parasite adaptation to the intracellular stages of its life cycle.

Animals↗

PubMed1999

Induction of a protective antibody response to foot and mouth disease virus in mice following oral or parenteral immunization with alfalfa transgenic plants expressing the viral structural protein VP1.

The utilization of transgenic plants expressing recombinant antigens to be used in the formulation of experimental immunogens has been recently communicated. We report here the development of transgenic plants of alfalfa expressing the structural protein VP1 of foot and mouth disease virus (FMDV). The presence of the transgenes in the plants was confirmed by PCR and their specific transcription was demonstrated by RT-PCR. Mice parenterally immunized using leaf extracts or receiving in their diet freshly harvested leaves from the transgenic plants developed a virus-specific immune response. Animals immunized by either method elicited a specific antibody response to a synthetic peptide representing amino acid residues 135-160 of VP1, to the structural protein VP1, and to intact FMDV particles. Additionally, the immunized mice were protected against experimental challenge with the virus. We believe this is the first report demonstrating the induction of a protective systemic antibody response in animals fed transgenic plants expressing a viral antigen. These results support the feasibility of producing edible vaccines in transgenic forage plants, such as alfalfa, commonly used in the diet of domestic animals even for those antigens for which a systemic immune response is required.

Administration, Oral↗

PubMed1999

An African swine fever virus ORF with similarity to C-type lectins is non-essential for growth in swine macrophages in vitro and for virus virulence in domestic swine.

An African swine fever virus (ASFV) ORF, 8CR, with similarity to the C-type lectin family of adhesion proteins has been described in the pathogenic isolate Malawi Lil-20/1. The similarity of 8CR to cellular and poxvirus genes associated with cell adhesion, cell recognition and virus infectivity suggested that 8CR may be of significance to ASFV-host cell interactions. Sequence analysis of the 8CR ORF from additional pathogenic ASFV isolates demonstrated conservation among isolates from both pig and tick sources. Northern blot analysis demonstrated 8CR mRNA transcription late in the virus replication cycle. A Malawi Lil-20/1 8CR deletion mutant (delta8CR) was constructed to analyse 8CR function further. The growth characteristics in vitro of delta8CR in porcine macrophage cell cultures were identical to those observed for parental virus. In domestic swine, delta8CR exhibited an unaltered parental Malawi Lil-20/1 disease and virulence phenotype. Thus, although well conserved among pathogenic ASFV field isolates, 8CR is non-essential for growth in porcine macrophages in vitro and for virus virulence in domestic swine.

African Swine Fever↗

PubMed1999

A new method for phenotyping red blood cells using microplates.

BACKGROUND AND OBJECTIVES: The supply of phenotyped red blood cells (RBC) for patients with several RBC antibodies presents a difficult task to hospital blood banks and regional blood centers. The aim of this study was to establish a low-cost typing system to allow extensive phenotyping of regular blood donors for clinically significant RBC antigens. MATERIALS AND METHODS: We developed a new buffer that greatly intensifies the antigen-antibody reaction and thus reduces the quantity of serum needed for phenotyping. The procedure was carried out on microplates. RESULTS: A total of 20,435 regular blood donors have been typed to date. For 752 units required for transfusion, 3,584 phenotyping tests were performed, validating the results by tube or gel typing methods; agreement was achieved in all cases. CONCLUSION: This technique seems adequate for phenotyping a large number of RBC units at very low cost, thus facilitating the availability of phenotyped blood.

Antibodies↗

PubMed1998

Expression of immunogenic glycoprotein S polypeptides from transmissible gastroenteritis coronavirus in transgenic plants.

The use of transgenic plants as vaccine production systems was described recently. We report on the immunological response elicited by two recombinant versions of the glycoprotein S from the swine-transmissible gastroenteritis coronavirus (TGEV) expressed in transgenic plants. Arabidoposis plants were genetically transformed with cDNAs constructs encoding either the N-terminal domain (amino acid residues 1-750) or the full-length glycoprotein S of TGEV, responsible for the neutralizing antibody induction against the virus, under the control of the cauliflower mosaic virus 35S (CaMV 35S) promoter. Genomic DNA and mRNA analyses of leaf extracts from transformed plants demonstrated the incorporation of the foreign cDNA into the arabidopsis genome, as well as their transcription. Expression of recombinant polypeptides were observed in most transgenic plants by ELISA using specific antibodies. Mice immunized with leaf extracts from transgenic plants developed antibodies that reacted specifically with TGEV in ELISA, immunoprecipitated the virus-induced protein, and neutralized the virus infectivity. From these results, we conclude that transgenic plants expressing glycoprotein S polypeptides may possibly be used as a source of recombinant antigen for vaccine production.

Animals↗

PubMed1998

Sexually transmitted infections in female sex workers: reduced by condom use but not by a limited periodic examination program.

BACKGROUND AND OBJECTIVES: The sexually transmitted diseases (STD) control program for female sex workers (FSW) in Lima, Peru, provided periodic serological tests for syphilis and cervical smears for gonococci, but not medication for STD or condoms. GOAL OF THIS STUDY: To assess program effectiveness. STUDY DESIGN: We assessed prevalence of current STD and serum antibody to STD pathogens in FSW in relation to program participation and to condom use. RESULTS: Program participation was not associated with significantly reduced rates of current gonococcal or chlamydial infections or Venereal Disease Research Laboratory (VDRL) titers > or =4 with reactive fluorescent treponemal antibody absorption (FTA-ABS). However, regardless of control program participation, consistent condom use was associated with reduced prevalence of gonorrhea, and with significantly reduced seroreactivity for FTA-ABS, C. trachomatis, anti-hepatitis B core (HBc), and anti-human T-lymphotropic virus type I. Anti-HBc was associated with years of receiving penicillin injections for syphilis prophylaxis. CONCLUSION: The scope, quality, and efficacy of STD control programs must be technically appropriate, well managed, and adequately financed. The safety of marginal programs warrants scrutiny.

Adult↗

PubMed1998

African swine fever virus NL gene is not required for virus virulence.

Previously, we described a highly conserved nonessential African swine fever virus (ASFV) right variable region gene, NL. Deletion of NL from the European pathogenic isolate E70 resulted in almost complete attenuation of the virus in domestic swine. To study gene function further, NL gene deletion mutants were constructed from two pathogenic African ASFV isolates, Malawi Lil-20/1 (Mal) and Pretoriuskop/96/4 (Pr4). Unexpectedly, both Mal (Mal-deltaNL) and PR4 (Pr4deltaNL) null mutants remained highly virulent when inoculated in swine. Mal-deltaNL exhibited a disease and virulence phenotype indistinguishable from its revertant, Mal-NLR, which caused 100% mortality. Mortality among Pr4deltaNL-infected animals was also high; however, a significant delay in onset of fever and viraemia and in time to death was observed. These data indicate that NL gene function is not required for ASFV virulence and that other yet-to-be identified viral determinants perform significant virulence functions in these African field isolates.

African Swine Fever Virus↗