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Biomedical subjects

I Blanco

Publications and source records attributed to I Blanco.

At least 73 records · Page 4Linked to original sources
PubMed1997

Chloroquine stabilizes pancreatic lysosomes and improves survival of mice with diet-induced acute pancreatitis.

Activation of digestive zymogens by lysosomal enzymes has been suggested as a triggering event in acute pancreatitis (AP). chloroquine (CQ), a weak base that accumulates in the lysosomes and increases their pH, can inhibit the activity of lysosomal enzymes. In the present study, we examined the effect of CQ on choline-deficient, ethionine-supplemented (CDE) diet-induced AP. CQ-diphosphate (15-50 mg.kg-1) or vehicle was given intraperitoneally at 0, 24, and 48 h to female CD1 mice that were fed with either normal diet or CDE diet. For mortality studies, animals were observed for 168 h. Serum and pancreas samples were collected from animals sacrificed 56 h after the start of the CDE diet. Treatment with CQ at 50 mg.kg-1 significantly (p < 0.05) improved the survival of mice with CDE diet-induced AP. In the normal pancreas, CQ decreased the specific activity of lysosomal enzymes cathepsin B1, beta-hexosaminidase, beta-glucuronidase, and acid phosphatase. In the pancreas with AP, CQ did not modify the activity of cathepsin B1, whereas it increased the latency of all enzymes. In conclusion, our results confirm the beneficial effect of CQ on survival of mice with CDE diet-induced AP and suggest that this effect of CQ may be due to its stabilizing action on lysosomes.

Acid Phosphatase↗
PubMed1997

Different routes of Ca2+ influx in NMDA-mediated generation of nitric oxide and arachidonic acid.

Nitric oxide and arachidonic acid act as inter- and intracellular messengers in the central nervous system. It is well known that the NMDA-mediated generation of nitric oxide and arachidonic acid is dependent on extracellular Ca2+. However, the role of voltage-dependent calcium channels (VDCCs) in this regard is poorly understood. We report here that NMDA-mediated nitric oxide production in striatal neuron cultures is blocked (80%) by the L-type VDCC antagonist nifedipine, but not by omega-conotoxin or omega-agatoxin IVA, antagonists of the N- and P-type VDCCs respectively. By contrast, none of the VDCC antagonists inhibited the NMDA-mediated release of arachidonic acid. These data indicate that permeation through different Ca2+ channels is responsible for the production of arachidonic acid and nitric oxide in striatal neurons.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1997

Comparison of Madin-Darby canine kidney cells (MDCK) with a green monkey continuous cell line (Vero) and human lung embryonated cells (MRC-5) in the isolation of influenza A virus from nasopharyngeal aspirates by shell vial culture.

We report a comparative study of the MDCK, Vero, and MRC-5 cell lines in the isolation of the influenza A (IA) virus. We studied 746 samples in which 63 IA viruses were isolated. The MDCK line displayed 100% sensitivity, the Vero line displayed 71.4%, and the MRC-5 displayed 57.1%. The MDCK line showed a statistically significant difference with respect to the Vero line (P = 0.001) and the MRC-5 line (P = 0.001). The quantitative sensitivity analysis showed the MDCK line to be superior to the other lines. It seems that the MDCK line is still one of the most recommendable for the isolation of the IA virus from respiratory samples.

Animals↗
PubMed1997

Reference values for retinol, tocopherol, and main carotenoids in serum of control and insulin-dependent diabetic Spanish subjects.

To establish reference ranges for use in clinical and epidemiological studies, we determined concentrations of retinol, alpha-tocopherol, beta-carotene, alpha-carotene, beta-cryptoxanthin, lutein, zeaxanthin, and lycopene in 450 Spanish control subjects and 123 Spanish patients with insulin-dependent diabetes mellitus (IDDM). Results were grouped according to sex, and samples were collected throughout the year. Concentrations of retinol were significantly lower and beta-carotene and alpha-carotene were higher in women than in men, both in controls and IDDM subjects, whereas beta-cryptoxanthin concentrations were higher only in control women. Conditional logistic regression analysis showed that retinol, beta-carotene, and lycopene were the variables associated with diabetes. In comparison with other populations, our controls showed, in general, ordinary concentrations of retinol, comparatively low beta-carotene and high beta-cryptoxanthin concentrations, and a relatively high alpha-tocopherol/ cholesterol ratio.

Adolescent↗
PubMed1997

[Comparison between the LLC-MK2 and MDCK cell lines in the isolation of parainfluenza virus from nasopharyngeal aspirates].

BACKGROUND: We report a prospective comparative study of the efficacy of three commercial cell lines in the isolation of the parainfluenza (PI) virus from nasopharyngeal samples. MATERIAL AND METHODS: In a 16 months period we studied all nasopharyngeal samples from patients with the suspension of a viral respiratory infection. The compared cell lines were LLC-MK2 and MDCK. All samples were processed by the shell-vial assay, incubated 3 days at 36 degrees C. The monolayers were stained by an indirect immunofluorescence assay with a monoclonal antibody against the nucleoprotein of the PI virus. RESULTS: In the study period 746 samples were analyzed, 46 PI virus were isolated, all belonged to the serotype 3. The LLC-MK2 cell line isolated 44 viruses (95.6%) and the MDCK cell line 41 (89.1%), no statistically significant differences were detected (p = 0.14) (qualitative sensitivity). In 84.8% of positive samples, the PI virus was isolated simultaneously in the two cell lines. Neither cell line isolated all PI virus. No statistical differences were detected in the quantitative sensitivity, although the LLC-MK2 cell line detected a slightly more samples with a low viral load (70.4 vs 68.2%). CONCLUSIONS: In view of the results obtained in this study, we believe that anyone of the two cell lines would be used in the isolation of the PI virus from respiratory samples.

Cell Line↗
PubMed1997

[Diagnosis of meningoencephalitis caused by herpes simplex virus type 1 using nested PCR in CSF samples].

BACKGROUND: The aim of this study was to achieve the early diagnosis of the neurologic alteration caused by the Herpes Simplex virus type 1 (HSV-1) with the nested PCR technique in CSF. PATIENTS AND METHODS: From January, 1994 to October, 1995, 140 CSF from 140 patients were studied in our laboratory. Ninety-five were diagnosed with viral meningoencephalitis (Group A) and 45 with other neurologic diseases (Group B). Nested PCR of HSV-1 and conventional viral cultures were carried out in all the cases. RESULTS: Laboratory diagnosis was achieved in 13 (13.68%) of Group A patients: in 12 (12.63%) HSV-1 genome was detected by nested PCR and in one patient adenovirus was isolated. In Group B, the HSV-1 was detected by nested PCR in 2 patients (4.44%). CONCLUSIONS: The results obtained demonstrate the usefulness of nested PCR in HSV-1 infection for the diagnosis of herpetic meningoencephalitis in initial stages of the disease, from a single CSF sample.

Herpes Simplex↗
PubMed1996

Modulation of N-methyl-D-aspartate receptor mediated arachidonic acid release by endogenous nitric oxide in cultured neurons.

Nitric oxide (NO) and arachidonic acid (AA) are believed to act as intra- and intercellular messengers in the central nervous system. Using primary cultures of striatal and hippocampal neurons, we have studied the effect of endogenous NO on N-methyl-D-aspartate (NMDA)-mediated AA release. Inhibition of NO synthesis with L-NG-nitroarginine (L-Noarg) produced a dose-dependent increase in NMDA-mediated AA release that was reversed by L-arginine. On the other hand, L-Noarg did not modify AA release produced by joint stimulation of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and phospholipase C-coupled metabotropic subtypes of glutamate receptors. Our results indicate that endogenously produced NO could modulate cellular events mediated by NMDA-induced AA release.

Animals↗
PubMed1996

Ascorbate increases the 1,25 dihydroxyvitamin D3-induced monocytic differentiation of HL-60 cells.

1,25 Dihydroxyvitamin D3 (calcitriol) induces differentiation of HL-60 leukemia cells. We studied the in vitro effect of a physiological concentration of ascorbate as potentiator of 1,25 dihydroxyvitamin D3 [(OH)2D3] activity by determining different markers of differentiation: nitroblue tetrazolium reduction, nonspecific esterase activity, and the expression of CD11b and CD14 surface antigens. Nitroblue tetrazolium reduction and nonspecific esterase activity increased up to 50% in the presence of both 1,25 (OH)2D3 plus 0.2 mM ascorbate (ASC), compared with (OH)2D3 as a unique agent. ASC also increased the expression of specific surface antigens (CD11b and CD14) during differentiation induced by 1,25 (OH)2D3, the effect being more pronounced after 48 hours of treatment with 10(-8) M 1,25 (OH)2D3. Furthermore, 1,25 (OH)2D3 alone increased intracellular cAMP level during differentiation, and the addition of ASC increased its concentration from 60 to 100% above the level reached with 1,25 (OH)2D3 as unique agent. ASC did not enhance the antiproliferative effect of calcitriol, suggesting that it only affects the ability of 1,25 (OH)2D3 to promote differentiation of HL-60 cells.

Ascorbic Acid↗
PubMed1996

Evaluation of a direct immunofluorescence assay, dot-blot enzyme immunoassay, and shell vial culture in the diagnosis of lower respiratory tract infections caused by influenza A virus.

We prospectively evaluated the efficacy of two commercial rapid methods for antigenic detection, a dot-blot enzyme immunoassay (EIA-DB) (Directigen FluA, Becton-Dickinson, USA) and a direct immunofluorescence assay (DIF) (Monofluokit Influenza A, Diagnostics Pasteur, France), compared with the shell-vial culture in the MDCK line, incubated 2 to 3 days and stained with the monoclonal antibody clone IA-52, the diagnosis of lower respiratory tract caused by Influenza A virus (IA). In the study period the presence of IA virus was detected in 59 of the 377 samples analyzed (15.7%). Only the SVC method detected all positive samples (100% sensitivity), being used as a reference method for comparison with the other techniques). The EIA-DB technique detected 50 cases (84.7%) and the DIF only 35 (59.3%). In nine (15.2%) cases the diagnosis was obtained only with the SVC method. The results of the comparison of the EIA-DB technique with SVC were: sensitivity 84.7%, specificity 100%, positive predictive value 100%, and negative predictive value 97.2%. The DIF technique gave values of 59.3%, 100%, 100%, and 92.9%, respectively. A statistically significant difference was observed between the sensitivity of the EIA-DB and the DIF method (p = 0.0001). In view of the results we recommended the use of the EIA-DB as a screening method when infection by the IA is suspected. But to obtain the maximum diagnostic yield all samples would be inoculated in a shell vial culture with the MDCK cell line.

Bacteriological Techniques↗
PubMed1996

Correlation between human cytomegalovirus quantitative p72 antigenemia and viremia.

OBJECTIVE: We present a prospective study of the correlation between the human cytomegalovirus (HCMV) quantitative antigenemia with monoclonal antibody to p72 protein (immediate-early antigen) and the number of infected cell foci detected in the shell-vial culture. A comparative study was made of the value of quantitative antigenemia (pp65 and p72) in 14 patients. RESULTS: The average value of the pp65 antigenemia was 195 pp65-positive PMNLs per 10(5) PMNLs (range 10-1000) and that of the p72, 21 p72-positive PMNLs per 10(5) PMNLs (range 0-120) (P < 0.001). The p72 antigenemia value represented 10.7% of the pp65 value (range 4.4-70%). A statistical correlation was observed between the total number of infected cell foci detected in the shell-vial culture and the total number of p72-positive PMNLs (P < 0.001), but not with the number of pp65-positive PMNLs (P = 0.4). A study of the number of infected cell foci detected in the shell-vial per 100000 PMNLs inoculated showed a statistical correlation with the value of the p72 antigenemia (P < 0.001). CONCLUSIONS: According to results, there seems to be a general population of PMNLs carrying viral particles which are defected by means of the pp65 monoclonal antibody, and a subpopulation carrying active and replicative viral particles which is detected with the p72 antibody. This last subpopulation would be responsible for the formation of infected cell foci in the shell-vial culture. However due to the technical difficulties presented by the routine performance of p72 antigenemia, we recommend the routine application of the quantitative shell-vial culture and the use of the number of infected cell foci x 100000 PMNLs inoculated as a parameter of replicative viral load for the diagnosis of infection and disease caused by HCMV.

Antibodies, Monoclonal↗
PubMed1996

Celiac plexus block as treatment for refractory pain related to sclerosing cholangitis in AIDS patients.

Sclerosing cholangitis may be a cause of refractory pain in patients infected with the human immunodeficiency virus. We performed celiac plexus block in three such patients with sever pain from sclerosing cholangitis and a poor response to conventional analgesia. The pain had been centered in the epigastrium and/or upper-right quadrant of the abdomen for 2, 10, and 15 weeks, respectively. Computed tomography-guided celiac plexus block with absolute alcohol and bupivacaine was performed. All three patients reported complete disappearance of the pain immediately after the procedure in two cases and 3 days later in the remaining patient. All patients were discharged free of pain and without analgesics and were followed up for 2, 8, and 11 months, respectively, without recurrence of pain. Celiac plexus block deserves further trial for the treatment of severe pain associated with sclerosing cholangitis in patients with acquired immunodeficiency syndrome. The quality of life of our three patients was considerably improved with this relatively simple procedure.

Abdominal Pain↗