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Biomedical subjects

J Sanchez

Publications and source records attributed to J Sanchez.

At least 37 records · Page 2Linked to original sources

Diverse BF recombinants have spread widely since the introduction of HIV-1 into South America.

OBJECTIVE: To describe the genetic diversity of HIV-1 in South America by full genome sequencing and analysis. METHODS: Purified peripheral blood mononuclear cell DNA from HIV-infected individuals in Argentina, Uruguay and Bolivia was used to amplify full HIV-1 genomes. These were sequenced using the ABI 3100 automated sequencer and phylogenetically analysed. RESULTS: Twenty-one HIV-1 strains from three South American countries, 17 of which were pre-screened by envelope heteroduplex mobility assay (HMA), were studied. Ten out of 10 HMA subtype F and four out of seven HMA subtype B strains were actually BF recombinants upon full genome analysis. Two BF recombinants from Argentina and two from Uruguay had the same structure, representing a new circulating recombinant form termed CRF12_BF(ARMA159). Twelve other BF recombinants had structures related to CRF12 but with additional segments of subtype B; each was unique. BF recombinants were temporally and geographically widespread, found as early as 1986-1987 in vertically infected Argentinian children and in Argentina, Uruguay, and Bolivia.

Adult↗

Promising early outcomes with a novel, complete steroid avoidance immunosuppression protocol in pediatric renal transplantation.

BACKGROUND: Corticosteroids have been a cornerstone of immunosuppression for four decades despite their adverse side effects. Past attempts at steroid withdrawal in pediatric renal transplantation have had little success. This study tests the hypothesis that a complete steroid-free immunosuppressive protocol avoids steroid dependency for suppression of the immune response with its accompanying risk of acute rejection on steroid withdrawal. METHODS: An open labeled prospective study of complete steroid avoidance immunosuppressive protocol was undertaken in 10 unsensitized pediatric recipients (ages 5-21 years; mean 14.4 years) of first renal allografts. Steroids were substituted with extended daclizumab use, in combination with tacrolimus and mycophenolate mofetil. Protocol biopsies were performed in the steroid-free group at 0, 1, 3, 6, and 12 months posttransplantation. Clinical outcomes were compared to a steroid-based group of 37 matched historical controls. RESULTS: Graft and patient survival was 100% in both groups. Clinical acute rejection was absent in the steroid-free group at a mean follow-up time of 9 months (range 3-13.7 months). Protocol biopsies in the steroid-free group (includes 10 patients at 3 months, 7 at 6 months, and 4 at 12 months) revealed only two instances of mild (Banff 1A) subclinical rejection (reversed by only a nominal increase in immunosuppression) and no chronic rejection. At 6 months the steroid-free group had no hypertension requiring treatment (P=0.003), no hypercholesterolemia (P=0.007), and essentially no body disfigurement (P=0.0001). Serum creatinines, Schwartz GFR, and mean delta height Z scores trended better in the steroid-free group. In the steroid-free group, one patient had cytomegalovirus disease at 1 month and three had easily treated herpes simplex stomatitis, but with no significant increase in bacterial infections or rehospitalizations over the steroid-based group. The steroid-free group was more anemic early posttransplantation (P=0.004), suggesting an early role of steroids in erythrogenesis; erythropoietin use normalized hematocrits by 6 months. CONCLUSIONS: Complete steroid-free immunosuppression is efficacious and safe in this selected group of children with no early clinical acute rejection episodes. This protocol avoids the morbid side effects of steroids without increasing infection, and may play a future critical role in avoiding noncompliance, although optimizing renal function and growth.

Adolescent↗

Efficient two-step chromatographic purification of penicillin acylase from clarified Escherichia coli ultrasonic homogenate.

A two-step chromatographic purification procedure from clarified Escherichia coli ultrasonic homogenate was evaluated. The capture step included immobilized metal affinity chromatography with Cu2+ as metal ion. Two elution methods were performed: 1 M NH4Cl and 0.01 M imidazole. Respectively, we obtained a different purification fold (16.5 to 3.15) and a similar result for the recovery of activity (90-99%). The best elution method was chosen for the procedure. The second step, hydrophobic interaction chromatography, gave a 3.8-fold purification with 77.7% of activity. The total procedure gave a 66-fold purification in relation to the initial crude extract with 70% for the recovery of activity and was performed without any conditioning step and at the same pH value.

Chromatography, Affinity↗

Risk perception, future land use and stewardship: comparison of attitudes about Hanford Site and Idaho National Engineering and Environmental Laboratory.

With the ending of the Cold War, the Department of Energy (DOE) is evaluating mission, future land use and stewardship of departmental facilities. This paper compares the environmental concerns and future use preferences of 351 people interviewed at Lewiston, Idaho, about the Hanford Site and Idaho National Engineering and Environmental Laboratory (INEEL), two of DOE's largest sites. Although most subjects lived closer to Hanford than INEEL, most resided in the same state as INEEL. Therefore their economic interests might be more closely allied with INEEL, while their health concerns might be more related to Hanford. Few lived close enough to either site to be directly affected economically. We test the null hypotheses that there are no differences in environmental concerns and future land-use preferences as a function of DOE site, sex, age and education. When asked to list their major concerns about the sites, more people listed human health and safety, and environmental concerns about Hanford compared to INEEL. When asked to list their preferred future land uses, 49% of subjects did not have any for INEEL, whereas only 35% did not know for Hanford. The highest preferred land uses for both sites were as a National Environmental Research Park (NERP), and for camping, hunting, hiking, and fishing. Except for returning the land to the tribes and increased nuclear storage, subjects rated all future uses as more preferred at INEEL than Hanford. Taken together, these data suggest that the people interviewed know more about Hanford, are more concerned about Hanford, rate recreational uses and NERP as their highest preferred land use, and feel that INEEL is more suited for most land uses than Handford. Overall rankings for future land uses were remarkably similar between the sites, indicating that for these stakeholders, DOE lands should be preserved for research and recreation. These preferences should be taken into account when planning for long-term stewardship at these two DOE sites.

Adult↗

The aegean sea oil spill in the Galicia Coast (NW Spain). I. Distribution and fate of the crude oil and combustion products in subtidal sediments.

The spatial distribution and fate of petrogenic and pyrogenic hydrocarbons in coastal sediments following the Aegean Sea oil spill (Galicia, NW Spain) was investigated through a detailed study of chemical markers. Alkanes and acyclic isoprenoid hydrocarbons were degraded within six months of the accident, but triterpane and sterane distributions were still detectable and were useful in monitoring, respectively, the oil source and weathering. Aromatic steranes were also useful source indicators, and oil degradation was clear from the decrease of certain alkyl dibenzothiophene and phenanthrene isomers. The pyrogenic polycyclic aromatic hydrocarbons produced in the tanker wreck were found more persistent than the petrogenic ones. However, the presence of the oil in the sediments was recognized even one year after the accident and was found particularly preserved in the subsurface layers.

Environmental Monitoring↗

Viability staining and terminal deoxyribonucleotide transferase-mediated dUTP nick end labelling of the mycelium in submerged cultures of Streptomyces antibioticus ETH7451.

Viability stain and terminal deoxyribonucleotide transferase-mediated dUTP nick end labelling (TUNEL) have been applied to submerged cultures of Streptomyces antibioticus ETH7451, the last technique after a suitable permeabilization treatment. Areas of dead mycelium can be clearly delineated by the viability stain within the network of hyphae which forms the mycelial masses characteristic of the submerged cultures. In addition, the TUNEL reaction shows that DNA fragmentation accompanies the death processes in the mycelium. These techniques permit the investigation of the influence of the medium and nutritional conditions on the viability of the cells. This has relevant biotechnological implications for the study of these important filamentous bacteria in the industrial fermentation processes. These techniques also allow a straight forward analysis of the physical and chemical reagents which provoke damage in Streptomyces DNA.

Culture Media↗

Role of extracellular calcium on the regulation of melatonin synthesis in the Syrian hamster pineal gland.

The role of extracellular calcium on the induction of cyclic AMP production, N-acetyltransferase (NAT) activity and melatonin synthesis, induced by forskolin, was investigated in Syrian hamster pineal glands. Pineals were removed from hamsters killed either in the first half of the normal dark period or late in the dark period. Forskolin immediately increased NAT activity and melatonin levels only when the glands were collected late in the dark period, while in the first half of the dark phase, hamster pineals responded to forskolin with an increase of NAT activity and melatonin production only after 6 hr of incubation. The absence of calcium prevented the induction of melatonin synthesis by forskolin only when glands were collected early in the dark phase and incubated for 6 hr. In the second half of the normal dark period removal of calcium markedly decreased NAT activity and melatonin levels in glands incubated with forskolin for either 4 or 6 hr. However, the absence of extracellular calcium had no significant effect on the induction of cyclic AMP production by forskolin in pineals collected either early in the dark period or late in the dark phase. These data indicate that at least part of the action of extracellular calcium is indirect and that it affects steps in the induction of melatonin synthesis beyond the accumulation of cAMP.

Animals↗

Prenatal diagnosis of 13q- syndrome in a fetus with holoprosencephaly and thumb agenesis.

Partial deletion of the long arm of one of the chromosomes 13 is an exceedingly rare condition. We report such a case in a 32-week fetus presenting with polyhydramnios, growth restriction and multiple structural defects including alobar holoprosencephaly, facial abnormalities, clubfoot, clinodactyly and thumb agenesis. Fetal blood sampling revealed a 46,XY, del(13)(q22 --> qter) abnormal male karyotype. Postmortem examination confirmed the prenatal findings and showed other manifestations of the syndrome. To our knowledge, this case represents the first in which the prenatal ultrasound detection of holoprosencephaly in association with distal limb abnormalities led to the prenatal diagnosis of the 13q- syndrome.

Adult↗

A comparison of HIV risk behaviors between new and long-term injection drug users.

The purpose of this study was to characterize the injection and sexual risk behaviors of a cohort of active drug injectors who have initiated injection within the past 4 years and to compare their behaviors with the risk behaviors of long-term injectors who have been injecting drugs since 1984. A stratified, network-based sample was used to recruit injection drug users from the streets in Miami-Dade, Florida. After screening for eligibility, which included a urine test to confirm current drug use, participants were administered a structured questionnaire that included basic demographic information, drug-use history, and HIV risk behavior practices. Both injector groups displayed a high level of HIV injection risk behavior. Although new initiates into injection demonstrated lower risk behavior than long-term injectors at the first injection episode, the current risk behavior between new and long-term injectors is similar.

Acquired Immunodeficiency Syndrome↗

Electrical nerve stimulation in the management of enterocutaneous low-output fistulas: a report of two cases.

Two patients with low-output enterocutaneous fistulas after surgery were treated with electrical nerve stimulation (ENS). Ultrasonography was useful for the application of this treatment method and for the charting of its progress. Fistula output diminished rapidly in both cases, and the closure of the track was achieved after several sessions of ENS. The procedure is simple and safe and is suggested as an option for the treatment of low-output enterocutaneous fistulas.

Aged↗

Functional antibody activity elicited by fractional doses of Haemophilus influenzae type b conjugate vaccine (polyribosylribitol phosphate-tetanus toxoid conjugate).

We evaluated the functional activities of antibodies, serum bactericidal activity (SBA), and immunoglobulin G (IgG) antibody avidity indices, using sodium thiocyanate (NaSCN) elution, elicited after vaccination with fractional doses of the Haemophilus influenzae type b conjugate (polyribosylribitol phosphate [PRP] conjugated to tetanus toxoid [PRP-T]) vaccine. A cohort of 600 infants from the Dominican Republic were randomized to receive one of three regimens of the PRP-T vaccine at ages 2, 4, and 6 months: full doses (10 microg of PRP antigen), one-half doses (5.0 microg), and one-third doses (3.3 microg) (J. Fernandez et al., Am. J. Trop. Med. Hyg. 62:485-490, 2000). Sixty serum samples, collected at age 7 months, with > or =2.0 microg of anti-PRP IgG per ml were randomly selected for avidity determinations. Geometric mean IgG concentrations were 13, 14, and 17 microg/ml for infants who received the full-dose (n = 19), one-half-dose (n = 19), and one-third-dose (n = 22) regimens, respectively. SBA geometric mean titers (1/dilution) were 85.0, 82.0, and 76.1 in sera from infants receiving the full-, one-half-, and one-third-dose regimens, respectively. Avidity indices (mean +/- standard error weighted average of NaSCN molar concentration x serum dilution factor) were 71.9 +/- 9.4, 123.6 +/- 26.8, and 150.9 +/- 24.9 for the full-, one-half-, and one-third-dose regimens, respectively. Upon comparison, the only significant difference (P = 0.024) found was a greater avidity index for sera from infants receiving the one-third-dose regimen than for sera from infants receiving the the full-dose regimen. We conclude that fractional doses elicit similar functional antibody activities in infants with > or = 2 microg of anti-PRP IgG per ml, corresponding to 89, 90, and 97% of infants receiving three doses of either the full concentration or one-half or one-third of the labeled concentration, respectively. This approach offers an alternative strategy for the prevention of H. influenzae type b disease in countries with limited resources.

Antibodies, Bacterial↗

Phase 2 trial of WR6026, an orally administered 8-aminoquinoline, in the treatment of visceral leishmaniasis caused by Leishmania chagasi.

There are no recognized orally administered treatments for any of the leishmaniases. The 8-aminoquinoline WR6026 is an orally administered analog of primaquine that cured 50% of patients with kala-azar in Kenya at a dose of 1 mg/kg/day for 28 days. A further phase 2, open-label, dose-escalating safety and efficacy study was performed for kala-azar in Brazil. Cure rates for Brazilian patients treated for 28 days were as follows: 1 mg/kg/day: 0 of 4 (0%); 1.5 mg/kg/day: 1 of 6 (17%); 2.0 mg/kg/day: 4 of 6 (67%); 2.5 mg/kg/day: 1 of 5 (20%); and 3.25 mg/kg/day: 0 of 1 (0%). Nephrotoxicity that was not anticipated from preclinical animal studies or from phase 1 studies was seen at 2.5 mg/kg/day in 2 patients and in the single patient administered 3.25 mg/kg/day. WR6026 demonstrated the unusual clinical features of lack of increased efficacy against Brazilian kala-azar with increased dosing above 2 mg/kg/day and toxicity that was not present in previous investigations.

Administration, Oral↗

Profile structures of TJ-II stellarator plasmas

Fine structures are found in the TJ-II stellarator electron temperature and density profiles, when they are measured using a high spatial resolution Thomson scattering system. These structures consist of peaks and valleys superimposed to a smooth average. Some irregularities remain in an ensemble average of discharges with similar macroscopic parameters such as line density, central temperature, and plasma current. They are found in all the magnetic configurations explored in plasmas heated by electron cyclotron waves. Their characteristics are shown and their possible origin is discussed.

Journal Article↗

Prevention of Haemophilus influenzae type b colonization by vaccination: correlation with serum anti-capsular IgG concentration.

Concentrations of serum anti-Haemophilus influenzae type b (anti-Hib) capsular polysaccharide (CPS) >/=0.15 and >/=1.0 microgram/mL are widely used as surrogates for protection against invasive Hib disease. However, the relationship between serum anti-Hib CPS following immunization and protection against colonization is not known, making it difficult to evaluate new Hib vaccines or combination vaccines. In the Dominican Republic, nasopharyngeal swabs were collected from 546 9-month-old infants who had received Hib conjugate vaccine at ages 2, 4, and 6 months and from 600 unvaccinated infants of the same age. The prevalence of Hib colonization was lower among vaccinated infants than among unvaccinated infants (0.9% vs. 2.3%). Among vaccinated infants, protection against colonization was significantly correlated with anti-Hib CPS concentrations >/=5 microgram/mL 1 month following the third dose of vaccine. These results suggest that the concentration of serum anti-Hib CPS needed for protection against colonization is greater than that needed for protection for invasive disease.

Antibodies, Bacterial↗

Emerging genetic diversity of HIV-1 in South America.

OBJECTIVE: Genotype determination and risk group analysis of HIV-1 infected individuals in selected regions of South America. DESIGN: Cross-sectional convenience sampling of HIV-1-positive individuals in Peru, Ecuador, Uruguay and Paraguay from March, 1994 through September, 1998. METHODS: HIV-1-positive subjects were identified through the national AIDS surveillance program in each country. A standardized questionnaire was used to obtain demographic, clinical and risk factor data on each study subject. Viral DNA was extracted from participants' peripheral blood mononuclear cells either directly or after co-cultivation. A nested PCR was used to obtain selected fragments of the envelope genes for genotyping by the heteroduplex mobility assay (HMA). A 600 bp sequence encompassing the V3 loop was sequenced from a selection of 23 of these samples for phylogenetic analysis and confirmation of HMA genotype. RESULTS: Among the 257 successfully genotyped HIV-1-positive samples, genotype B was found in 98.3% (228/232) of those obtained from subjects in Peru, Ecuador, and Paraguay. In contrast, 56% (14/25) of the samples from Uruguay were genotype F, and the remainder were genotype B. Genotype F was detected for the first time in Peru (2/224) and Paraguay (1/4), and genotype A for the first time in Peru (1/224). Phylogenetic analysis confirmed the genotype identified by HMA in the 23 samples sequenced. There was no detectable genetic clustering of HIV-1 within the different high-risk groups or geographic locations. CONCLUSIONS: These findings verify and extend the presence of several different HIV-1 genotypes in South America.

Amino Acid Sequence↗

Molecular analysis of lineage-specific chimerism and minimal residual disease by RT-PCR of p210(BCR-ABL) and p190(BCR-ABL) after allogeneic bone marrow transplantation for chronic myeloid leukemia: increasing mixed myeloid chimerism and p190(BCR-ABL) detection precede cytogenetic relapse.

We studied lineage-specific chimerism and minimal residual disease (MRD) in sequential posttransplant samples from 55 patients who underwent unmanipulated (n = 44) or partially T-cell-depleted (n = 11) allogeneic bone marrow transplantation (BMT) for chronic myeloid leukemia (CML). Chimerism was assessed by polymerase chain reaction (VNTR [variable number of tandem repeats]-PCR) analysis in highly purified CD19+, CD3+, CD15+, and CD56+ cell fractions, whereas MRD was investigated in whole blood by reverse transcriptase-PCR (RT-PCR) of both p210(BCR-ABL) and p190(BCR-ABL) hybrid transcripts. Of 55 patients, 14 (including 6 T-cell-depleted patients) had cytogenetic relapse at 5-80 months and progressed to hematologic relapse, while 41 patients remained in prolonged cytogenetic remission 12-107 months post-BMT. Before leukemia recurrence, patients in the relapse group showed a consistent evolution pattern sequentially featured by persistent p210(BCR-ABL) positivity, increasing mixed chimerism (MC) in myeloid cells, p190(BCR-ABL) positivity, and, finally, cytogenetic relapse. Myeloid MC preceded cytogenetic relapse by 2-12 months, whereas p190(BCR/ABL) was detected 1-6 months prior to cytogenetic relapse in 11 patients and concomitant with cytogenetic relapse in 3 patients. In the remission group, all patients invariably tested negative for p190(BCR-ABL); 10 patients tested positive for p210(BCR-ABL) at variable time-points but showed persistent full donor chimerism (DC), whereas 31 patients tested p210(BCR-ABL) negative and displayed full DC or transient MC due to the persistence of recipient T cells. Two patients in the relapse group were successfully reinduced into molecular remission with donor lymphocyte infusion. Sequential molecular analysis after such treatment showed the inverse pattern to that observed prior to relapse, ie, progressive disappearance of p190(BCR-ABL) transcripts, conversion of myeloid chimerism to donor type, and, finally, p210(BCR-ABL) negativity. We conclude that lineage-specific chimerism and p190(BCR-ABL) messenger RNA (mRNA) analyses contribute a better characterization of CML evolution after BMT and enable early identification of patients at the highest risk of relapse. (Blood. 2000;95:2659-2665)

Adolescent↗

Lipoxygenase pathway in olive callus cultures (Olea europaea).

Stimulation of the lipoxygenase pathway in olive fruit initiates a cascade of reactions that begins with the regio- and stereospecific di-oxygenation of polyunsaturated fatty acids containing a cis, cis-1,4 pentadiene moiety. Later products of the pathway include volatiles that influence the organoleptic properties of harvested olive oil. In this study, we have investigated lipoxygenase activity in olive callus cultures, and found that there is evidence of several isoforms of the enzyme with different pH optima and substrate specificities. Endogenous lipoxygenase activity was detected throughout the growth cycle of olive callus, particularly during the log phase of growth, suggesting that olive lipoxygenases are intimately involved in growth. The most prominent lipoxygenase activity in tissue cultures was found to be soluble but significant activities were detected in the plastid fraction. In addition, hydroperoxide lyase (HPL) activity was measured in the calli; both 13- and 9-HPL activities were found which were particulate.

Aldehyde-Lyases↗

Isolation and hypoglycemic activity of 5, 7,3'-trihydroxy-3,6,4'-trimethoxyflavone from Brickellia veronicaefolia.

Hypoglycemic activity-guided fractionation together with chemical analysis led to the isolation of one flavone (5, 7,3'-trihydroxy-3,6,4'-trimethoxyflavone) from the chloroform extract of the leaves of Brikkellia veronicaefolia. Identification was based on spectroscopic methods. The isolated flavone was tested for hypoglycemic activity in normal and alloxan-diabetic CD1 mice (25-30 g) were administered in doses of 10, 25 and 50 mg/kg body weight. The blood glucose levels were determined before and 1.5, 3, 4.5 and 24 hours after drug administration. The results showed that the flavone produces a significant hypoglycemic effect in normal as well as in diabetic mice. Comparison was made between the action of the flavone and a known hypoglycemic drug as tolbutamide (50 mg/kg). The flavone was found to be slow and less effective than tolbutamide.

Alloxan↗