Search PubMed⌕ Search

Biomedical subjects

A Garcia

Publications and source records attributed to A Garcia.

At least 217 records · Page 12Linked to original sources

Treatment with imipenem of experimental endocarditis caused by penicillin-resistant Streptococcus sanguis.

The efficacy of imipenem alone or in combination with gentamicin for the treatment of experimental endocarditis in rabbits caused by a penicillin resistant strain of Streptococcus sanguis was compared to that of gentamicin and penicillin G. In vitro, both beta-lactams were bactericidal in action and the combination of gentamicin and imipenem was synergistic after 4 h whereas penicillin G and gentamicin showed synergy after 8 h. Antibiotic treatment was started two days after initiating infection and groups of 20 animals were treated for 5 days with imipenem alone or in combination with gentamicin or with gentamicin and penicillin G. Two of 20 rabbits which received no therapy died whereas all other rabbits survived the treatment period. Blood cultures became negative in 19 rabbits on the second day of treatment with imipenem and gentamicin and sterilization of the vegetations was achieved in 12 animals. In contrast, eight animals given the alternative treatments remained bacteraemic and viable organisms were recovered from the vegetations in every case. However, bacterial counts and vegetation weights only differed significantly between treated animals and controls but not between the different treatment groups. Never the less, these data suggest that combined treatment with imipenem and gentamicin might provide an effective regimen for patients with endocarditis caused by penicillin resistant S. sanguis.

Animals↗

Myocardial uptake of antimyosin antibody in idiopathic dilated cardiomyopathy and its relation to functional and morphological parameters.

Monoclonal 111In antimyosin (AMS) uptake indicates the presence of ongoing myocyte damage. In idiopathic dilated cardiomyopathy (IDC), there is diffuse myocyte damage. We have attempted to find a correlation between AMS uptake and functional myocardial parameters. With this purpose in mind, we studied two groups of subjects: group 1 comprised 19 subjects with IDC and group 2 comprised 6 control subjects. In all subjects, an antimyosin scan was performed. Among the subjects with IDC, two-dimensional echocardiography was carried out to determine the left ventricular ejection fraction (LVEF) and left ventricular dimensions, and a gated blood pool study was undertaken to assess the LVEF at rest and end-diastolic and end-systolic volumes. Three months later, repeat antimyosin scintigraphy and equilibrium gated blood pool were performed on 13 of the patients. The mean heart to lung (H/L) ratio in the IDC subjects was 1.82 +/- 0.25 (range 1.42-2.25), a value significantly higher than that obtained in the controls: 1.41 +/- 0.12 (range 1.26-1.58) (P < 0.001). Linear regression analysis did not find a statistically significant correlation between H/L and gated blood pool or echocardiography measures. No marked changes in ejection fraction and antimyosin uptake were found between baseline and follow-up studies. Subjects with IDC have a high incidence of positive antimyosin scans, but antimyosin uptake is not related to any functional or morphological parameters.

Antibodies, Monoclonal↗

Characterization of the trehalose 6,6'-dimycolate surface monolayer by scanning tunneling microscopy.

The toxicity of trehalose 6,6'-dimycolate (TDM), a glycolipid of mycobacteria, requires presentation as a surface monolayer. Our model of the structure of the TDM monolayer was confirmed and extended by scanning tunneling microscopy. It consists of linear arrays with a periodicity of approximately 90 A (9 nm) that clustered in groups of four to form secondary structures with a periodicity of 360 A (36 nm).

Cord Factors↗

HLA-B alleles and complotypes in Mexican patients with seronegative spondyloarthropathies.

OBJECTIVES: To analyse major histocompatibility complex (MHC) haplotypes in Mexican mestizo patients with seronegative spondyloarthropathies (SSpA) and normal controls, to discover if there are other antigens, besides B27, in the HLA region that might show association with the disease. METHODS: The study included 100 Mexican mestizo patients with SSpA and 200 of their first degree relatives. These groups were compared with 85 ethnically matched controls. The class I and class III MHC antigens were obtained by standard methods. The significance of differences between patients and controls was tested by chi 2 analysis; linkage disequilibrium among the different alleles in each haplotype was estimated by computing delta values. RESULTS: We found a significantly increased frequency of the HLA-B27 antigen (pcorr. = 1 x 10(-5), odds ratio (OR) = 33.4, 95% confidence interval (CI) = 9.3-142.0). In the group of 45 SSpA patients negative for the B27 antigen, independent increased frequencies of HLA-B49 antigen (pcorr. = 0.03, OR = 6.5, 95% CI = 1.5-32.8)) and the FC31 complotype (pcorr. = 0.04, OR = 3.7, 95% CI = 1.2-11.1) were found. Significant delta values were obtained for the [B27;SC30] haplotype (p = 0.0005) but not for haplotypes marked by the FC31 complotype. HLA-B antigens on the homologous chromosome in B27 positive patients were mainly HLA-B51 (18%) and HLA-B60 (16%); however, the observed genotypes B27/B51 and B27/B60 were not significantly different than expected from the allele frequencies alone. CONCLUSIONS: These data suggest that in Mexicans additional genes within the MHC region besides the HLA-B27 antigen, might be related to the genetic susceptibility for developing SSpA. Relevant antigens included the HLA-B49 and the FC31 complotype.

Adult↗

Diagnosis of polyomavirus-induced hepatic necrosis in psittacine birds using DNA probes.

Liver sections from 32 psittacine birds with multifocal to coalescing hepatocellular necrosis were examined to determine the cause of disease. Avian polyomavirus (APV) infection (19 of 32 birds), bacterial hepatitis (5 of 32 birds), and chlamydiosis (3 of 32 birds) were major causes of hepatic disease. The presence of APV inclusions or nucleic acid was demonstrated using hematoxylin and eosin (HE) staining, DNA in situ hybridization, and DNA amplification with Southern blotting. Amphophilic intranuclear inclusions, suggestive of APV infection, were observed in HE-stained liver sections from 5 of 32 birds. Hepatocellular karyomegaly was present in liver tissues from 10 birds (5 birds with typical APV inclusions and 5 birds without discernable inclusions). DNA in situ hybridization recognized intranuclear APV nucleic acid in liver sections of 18 of 32 birds. DNA amplification with Southern or dot blots also identified APV nucleic acid in processed, paraffin-embedded livers of 18 of 32 birds. This study demonstrates that acute APV infection is a frequent cause of multifocal to coalescing hepatocellular necrosis in psittacine birds. Furthermore, APV infection is best diagnosed using DNA probes, especially when typical intranuclear inclusions are not observed microscopically.

Animals↗

Critical evaluation of the present experience on microsurgical sperm aspiration.

Microsurgical epididymal sperm aspiration (MESA) and IVF techniques are indicated in those cases of bilateral congenital absence of the vas deferens (BCAVD) and in cases of epididymal obstruction (EO) of other aetiologies were vasoepididymostomy is not possible or has failed. Microsurgical deferential sperm aspiration (MDSA) is indicated in those cases where spermatogenesis and epididymal permeability are conserved but spermatozoa, for different causes, cannot reach the ejaculate and other therapies have failed. We have performed this procedure in 70 patients by two different human reproduction teams. Both teams achieved very similar results regarding fertilization rate: 5.5 and 7.2% in cases of BCAVD, 6.5 and 8.8% in cases of EO and 33.3 and 26.7% in cases of non-epididymal pathology (NEP). However, the fertilization rate per patient presents differences between both teams: 27 and 44.4% in BCAVD, 31 and 11% in EO and 80 and 100% in NEP. In spite of a better fertilization rate per patient in team two, it was in the other where pregnancies were achieved (7% in BCAVD, 15% in EO and 20% in NEP). One pregnancy in EO group was achieved by intracytoplasmatic sperm injection. The pregnancy rates per transfer were 25% in BCAVD, 50% in EO and 25% in NEP.

Epididymis↗

Lymphocyte subpopulations in patients with primary fibromyalgia.

OBJECTIVE: Fibromyalgia (FM) is a clinical entity of unknown etiology frequently diagnosed in rheumatology. The potential involvement of the immune system in its pathogenesis has been suggested. Studies of abnormal T cell subpopulations often have been inconclusive. We attempted to clear this point by comparing lymphocyte subpopulations, including some of the newer activation markers, in patients with FM and healthy controls. METHODS: Sixty-five patients with FM and 56 healthy controls were studied. Flow cytometry was used as a quantification technique to measure lymphocyte subpopulations, CD3 (T cells), CD19 (B cells), CD16 (natural killer cells), CD4 (T helper/inducer cells), CD8 (T cytotoxic/suppressor cell), CD25 (interleukin 2 receptor), CD69 (activation inducer molecule marker), CD71 (transferrin receptor) and CD54 (ICAM-1); CD4/CD8 ratios were also estimated. RESULTS: The number of T cells expressing activation markers CD69 and CD25 was decreased in patients with FM; the other subpopulations were similar in patients and controls. CONCLUSION: Our results suggest a defect in T cell activation in patients with FM.

Adolescent↗

Pericardial and myocardial localization of antimyosin in a case of acute myocarditis.

A 60-yr-old immunodepressed woman presented with acute perimyocarditis and cardiac tamponade complicating meningococcic infection. We had the opportunity to study her cardiac condition by injecting antimyosin during the acute phase. Images at 48 hr showed unexpected findings, with obvious localization of the tracer in the pericardial fluid, as well as myocardial uptake. Possible mechanisms for pericardial activity are discussed.

Antibodies, Monoclonal↗

Histologic predictive factors. Therapeutic impact in vulvar cancer.

In 59 cases of vulvar invasive squamous cell carcinoma treated with radical vulvectomy and inguino-femoral lymphadenectomy, seven histologic parameters were evaluated to establish their predictive value in the development of lymph node metastasis. The most significant was vascular involvement, observed in 86% of cases with lymph node metastases (P < .000004). Depth of stromal invasion and tumor thickness were highly significant, with P < .008 and < .007, respectively, with 0% lymph node metastases in tumors thinner than 1 mm and 62% and 60%, respectively, in those thicker than 5 mm. The growth pattern correlated with lymph node metastases but was not statistically significant. The histologic grade correlated with positive lymph nodes, with P < .04. The amount of keratin (P < .91) was not related. These histologic factors allow the identification of patients with a lower risk of developing lymph node metastases and in whom conservative surgery on the vulva and inguino-femoral lymph nodes is feasible.

Adult↗

Long-term stability of baculoviruses stored under various conditions.

The long-term stability of baculoviruses under various storage conditions was investigated. The results showed that the titers of baculovirus working stocks were significantly reduced after the viruses had been stored in polypropylene tubes for several months or a year at 4 degrees C. The virus was not lost by sticking to the plastic tubes, because the same results were obtained when the virus was stored in glass tubes. The major factor contributing to the loss of viral infectivity was exposure to light. Virus stocks stored at temperatures ranging from -85 degrees to 37 degrees C were quite stable, as long as they were protected from light. Finally, viral infectivity was not significantly reduced by multiple freeze-thaw cycles. These results demonstrated that baculovirus working stocks may be stored frozen or refrigerated with little long-term loss of infectivity, as long as they are protected from light.

Baculoviridae↗

Influence of different signal peptides and prosequences on expression and secretion of human tissue plasminogen activator in the baculovirus system.

Foreign secretory pathway proteins are often produced in surprisingly low amounts in the baculovirus/insect cell expression system. One possible reason for this is that heterologous signal peptides might be inefficiently recognized by the insect cell protein translocation machinery. This idea was supported by a recent study showing that secretion of a plant protein in the baculovirus system was enhanced when its signal peptide was replaced with an insect-derived signal peptide (Tessier, D. C., Thomas, D. Y., Khouri, H. E., Laliberte, F., and Vernet, T. (1991) Gene (Amst.) 98, 177-183). We have extended these observations by measuring the effects of different signal peptide and signal peptide-prosequence combinations on baculovirus-mediated expression and secretion of human tissue plasminogen activator (t-PA). Replacement of the native prepropeptide with signal peptides from a lepidopteran insect secretory protein (cecropin B), a major baculovirus structural glycoprotein (64K), or an abundant, highly conserved lumenal protein of the rough endoplasmic reticulum (GRP78/BiP, a 78-kDa glucose-regulated protein/immunoglobulin heavy chain-binding protein), had no significant effect on t-PA expression or secretion. The same results were obtained with the signal peptide from honeybee prepromellitin, which was able to enhance secretion of plant propapain (Tessier et al., 1991 (above)). Similar results were obtained when heterologous signal peptides were combined with the native prosequence or when the intact cecropin B preprosequence was used. Translational initiation at an upstream, in-frame ATT, which could functionally inactivate any signal peptide, did not explain the low efficiency of t-PA secretion. Finally, deletion of the native signal peptide, prosequence, or both, failed to increase t-PA production. These results showed that insect-derived signal peptides and/or prosequences cannot always enhance the expression and/or secretion of foreign secretory pathway proteins in the baculovirus system. They also suggested that the inability of insect cells to recognize the processing signals in human t-PA efficiently is probably not the major factor preventing its high level production in this system.

Amino Acid Sequence↗

DNA adduct-induced stabilization of slipped frameshift intermediates within repetitive sequences: implications for mutagenesis.

Chemical carcinogens such as the aromatic amide 2-acetylaminofluorene (AAF) are known to induce -1 frameshift mutation hotspots at repetitive sequences. This mutagenesis pathway was suggested to involve slipped intermediates formed during replication. To investigate the stability and structure of such intermediates we have constructed DNA duplexes containing single AAF adducts within a run of three guanine residues. The strand complementary to that bearing the AAF adducts contained either the wild-type sequence (homoduplexes) or lacked one cytosine directly opposite the run of guanines containing the AAF adduct and thus modeled the putative slipped mutagenic intermediates (SMIs). The melting temperature of AAF-modified homoduplexes or the unmodified SMI was reduced by approximately 10 degrees C relative to the unmodified homoduplex. Surprisingly, AAF adducts stabilized the SMIs as evidenced by an increase in melting temperature to a level approaching that of the unmodified homoduplex. The chemical probes hydroxylamine and bromoacetaldehyde were strongly reactive toward cytosine residues opposite the adduct in AAF-modified homoduplexes, indicating adduct-induced denaturation. In contrast, no cytosine reactivities were observed in the AAF-modified SMIs, suggesting that the two cytosines were paired with unmodified guanines. Use of diethyl pyrocarbonate to probe the guanine residues showed that all three guanines in the unmodified SMI adopted a transient single-stranded state which was delocalized along the repetitive sequence. However, when an AAF adduct was present, reduced diethyl pyrocarbonate reactivity at guanines adjacent to the adduct in AAF-modified SMIs reflected localization of the bulge to the adducted base. Our results suggest that AAF exerts a local denaturing and destabilizing effect within the homoduplex which is alleviated by the formation of a bulge. The stabilization by the AAF adduct of the SMIs may contribute to the dramatic increase in -1 frameshift mutation frequency induced by AAF adducts in repetitive sequences.

2-Acetylaminofluorene↗

Transmissible gastroenteritis in pigs in south east Spain: prevalence and factors associated with infection.

In the Murcia region of south east Spain, epidemics of transmissible gastroenteritis-like disease have occurred in pigs every three years since 1980. In 1987 a sero-epidemiological survey was conducted to estimate the prevalence and spread of transmissible gastroenteritis virus (TGEV) infection among breeding pigs and farms, and to determine any association between the infection and herd size and geographical zone. The Murcia region was divided into four geographical zones and the farms were classified by size into four categories. The random sample was statistically representative of both the breeding stock and the farms in each geographical zone, and the analyses involved 6000 breeding pigs from 480 farms. The immunological techniques employed were the indirect ELISA and the blocking ELISA. The prevalence of seropositive breeding pigs was 1.27 +/- 0.28 per cent and the prevalence of infected farms was 5 +/- 1.94 per cent. On the infected farms, the prevalence of seropositive pigs ranged from 5 to 60 per cent. The infection occurred throughout the Murcia region, but a significant association (P < 0.01) was observed between the geographical zone and the prevalence of TGEV infection. Herds with > or = 50 breeding pigs had a greater prevalence (P < 0.01) of TGEV infection.

Animals↗

Differential response to frameshift signals in eukaryotic and prokaryotic translational systems.

The genomic RNA of beet western yellows virus (BWYV) contains a potential translational frameshift signal in the overlap region of open reading frames ORF2 and ORF3. The signal, composed of a heptanucleotide slippery sequence and a downstream pseudoknot, is similar in appearance to those identified in retroviral RNAs. We have examined whether the proposed BWYV signal functions in frameshifting in three translational systems, i.c. in vitro in a reticulocyte lysate or a wheat germ extract and in vivo in E. coli. The efficiency of the signal in the eukaryotic system is low but significant, as it responds strongly to changes in either the slip sequence or the pseudoknot. In contrast, in E. coli there is hardly any response to the same changes. Replacing the slip sequence to the typical prokaryotic signal AAAAAAG yields more than 5% frameshift in E. coli. In this organism the frameshifting is highly sensitive to changes in the slip sequence but only slightly to disruption of the pseudoknot. The eukaryotic assay systems are barely sensitive to changes in either AAAAAAG or in the pseudoknot structure in this construct. We conclude that eukaryotic frameshift signals are not recognized by prokaryotes. On the other hand the typical prokaryotic slip sequence AAAAAAG does not lead to significant frameshifting in the eukaryote. In contrast to recent reports on the closely related potato leafroll virus (PLRV) we show that the frameshifting in BWYV is pseudoknot-dependent.

Base Sequence↗

Association of HLA-DR4-Dw15 (DRB1*0405) and DR10 with rheumatoid arthritis in a Spanish population.

OBJECTIVE: To analyze the associations of HLA class II antigens with rheumatoid arthritis (RA) in a Spanish population. METHODS: We used DNA oligotyping to determine DR types, DQA1 and DQB1 alleles, and DR4 variants in 70 unrelated seropositive RA patients and 189 healthy controls living in Spain. RESULTS: A significantly higher frequency of DR4 was seen in RA patients compared with controls (relative risk [RR] = 2.40). The DR10 specificity correlated most strongly with disease susceptibility (RR = 3.84). A significant decrease in the frequency of DR7 was observed in the RA patients (RR = 0.48). DR4-Dw15 (DRB1*0405) was found to be the unique DR4 allele associated with RA (RR = 4.27, P < 0.05), whereas Dw4 (DRB1*0401) and Dw14 (DRB1*0404/0408) showed no association, and both Dw10 (DRB1*0402) and Dw13 (DRB1*0403/0407) were negative risk factors for the disease. Approximately one-third of the cases of RA could not be explained by the "shared epitope" hypothesis. Investigation of the DQ alleles associated with DR4 showed that the haplotype Dw15-DQ8 (DRB1*0405-DQB1*0302) was a susceptibility factor for RA (RR = 6.36, P < 0.05). CONCLUSION: Our results suggest that HLA class II alleles involved in RA susceptibility can vary among different Caucasian populations.

Alleles↗