Search PubMed⌕ Search

Biomedical subjects

J Cruz

Publications and source records attributed to J Cruz.

At least 55 records · Page 3Linked to original sources
PubMed1999

Successful PBSC mobilization with high-dose G-CSF for patients failing a first round of mobilization.

PBSC are the preferred source of stem cells for autologous transplantation. However, regardless of the mobilization procedure used, 10%-20% of patients fail to collect an adequate number to ensure prompt engraftment. There is as yet no standard mobilization procedure for patients who fail a first mobilization attempt. Here, we describe a highly efficient strategy to obtain an adequate number of stem cells for patients who failed a first mobilization attempt. Seventy-four patients with various hematologic malignancies underwent initial mobilization with various regimens including hematopoietic growth factors with or without chemotherapy. In 72% of patients, > or =2 x 10(6) CD34+ stem cells/kg were collected in the initial mobilization attempt, and patients engrafted in a median of 10 days for neutrophils and 12 days for platelets. Eighteen patients failed to mobilize adequate numbers of stem cells, defined as the inability to collect 0.2 x 10(6) CD34+ stem cells/kg/day in the first 2-3 days. These patients had their apheresis halted. Patients were immediately given G-CSF (32 microg/kg/day) for 4 days as a second attempt at mobilization. Eighty-eight percent of these patients achieved the target of > or =2 x 10(6) CD34+ cells/kg, with a median duration of apheresis of 5 days (including the first and second mobilizations). The mean CD34+ cells/kg/day increased after administration of high-dose G-CSF from 0.16 after the first mobilization attempt to 0.61 (p = 0.0002) after the second mobilization. All patients engrafted in a median of 11 and 13 days for neutrophils and platelets, respectively. We conclude that patients whose apheresis yield is <0.4 x 10(6) CD34+ cells/kg after the first two apheresis collections can be successfully mobilized if high-dose G-CSF is administered immediately and continued until achieving > or =2 x 10(6) CD34+ stem cells/kg.

Adult↗
PubMed1998

Putative dengue virus receptors from mosquito cells.

Dengue viruses are arthropod-borne, single-stranded RNA viruses. Aëdes aegypti and Aëdes albopictus are the principal vectors. In order to understand the molecular basis of dengue virus infections we explored the biochemical identity of dengue-2 (DEN-2) virus receptors in the Aëdes albopictus-derived cell line C6/36. We show here that DEN-2 interacts with two major polypeptides of 80 and 67 kDa. Polyclonal anti-C6/36 membrane antibodies block DEN-2 binding to intact C6/36 monolayers as well as to membrane extracts. Our results strongly suggest that the identified polypeptides are part of the DEN-2 receptors.

Aedes↗
PubMed1998

The biology and ethics of banking fetal liver hematopoietic stem cells for in utero transplantation.

BACKGROUND/PURPOSE: Transplantation of fetal liver hematopoietic stem cells (HSCs) in utero has the potential to treat a variety of hematologic, immunologic, and metabolic diseases. One prerequisite for broad clinical application is the establishment of a bank of fetal liver HSC tissue. The authors describe their methods for processing fetal liver free of known human pathogens while maximizing HSC activity after cryopreservation. METHODS: The authors developed a protocol that separates the abortion decision from the donation decision and preserves confidentiality between donor and recipient. Human fetal livers (12 to 14 weeks' gestation) were procured from aborted specimens and the light-density hematopoietic cells isolated by density centrifugation. Total viable cell count increased with gestational age and averaged from 4.36 x 10(7) cells for 12-week livers to 2.0 x 10(8) cells for 14-week livers. RESULTS: Flow cytometric analysis demonstrated the presence of early progenitors in fresh and thawed specimens and a low number of T cells in each group. The functional capacity of fetal liver progenitors was assessed with colony-forming assays before and after cryopreservation. Thawed specimens showed an average 63% recovery rate for the high-proliferative potential colony-forming cells, a primitive subset of progenitors thought to include HSC. However, the more mature fraction of low-proliferative potential colony-forming cells had a recovery rate of only 35%. These data suggest that fetal liver HSC maybe more resistant to the detrimental effects of cryopreservation than mature progenitors. The fetal liver was screened for bacterial, fungal, and viral contaminates and the serum from donor mothers was screened for human immunodeficiency virus (HIV), hepatitis A, B, and C, human T-cell lymphoma virus (HTLV I/II), rapid plasma reagent (RPR), cytomegalovirus (CMV), and toxoplasmosis IgM. The bacterial contamination rate was 14% (n = 28). The maternal serum was positive for CMV in 78% of cases, and positive for hepatitis C in 0.7% of cases (n = 28). However, all fetal liver specimens were culture negative for CMV. CONCLUSIONS: These findings demonstrate that human fetal liver HSCs can be procured ethically and processed to ensure a safe graft with a small number of T-cells, and a high yield of progenitors after cryopreservation. A bank of fetal liver HSC will prove useful in treating a variety of genetic diseases before birth by in utero HSC transplantation.

Cryopreservation↗
PubMed1998

Quality of life and psychological distress of bone marrow transplant recipients: the 'time trajectory' to recovery over the first year.

The purpose of this study was to measure the trajectory of psychosocial recovery over the first year after bone marrow transplantation (BMT). BMT patients were assessed at baseline (n = 86), hospital discharge (n = 74), 100 days (n = 64) and at 1 year (n = 45). Participants completed the Functional Assessment of Cancer Therapy-Bone Marrow Transplant Scale (FACT-BMT), the Profile of Mood States Total Mood Disturbance Scale (POMS-TMDS), the Medical Outcomes Social Support Survey (MOS-SSS), the Center for Epidemiologic Studies-Depression (CES-D) scale screener, a performance Status Rating Scale (PSR), and an interview questionnaire. The recovery trajectory in this patient population showed three distinct trends. The trajectory for distress was linear and improved over time with approximately 20% of patients continuing to have psychological distress at 1 year. Secondly, the trend for overall quality of life was parabolic, worsening at discharge, then improving at 100 days and at 1 year. However, there were individual areas of deficit at follow-up, eg fatigue, even while overall quality of life mean scores improved. Thirdly, the trend for patient concerns over time was linear and worsening. These recovery trajectories suggest psychosocial interventions before and after BMT that may prepare patients for increasing and worsening concerns even as physical well-being improves.

Adult↗
PubMed1998

The first decade of continuous monitoring of jugular bulb oxyhemoglobinsaturation: management strategies and clinical outcome.

OBJECTIVE: To comparatively assess outcome of patients undergoing monitoring and management of cerebral extraction of oxygen along with cerebral perfusion pressure vs. outcome of patients undergoing monitoring and management of cerebral perfusion pressure alone in severe acute brain trauma. DESIGN: Prospective, interventional study. SETTING: Intensive care unit of a university hospital. PATIENTS: Adults (n = 353) with severe acute brain trauma. A group of 178 patients underwent continuous monitoring and management of cerebral extraction of oxygen and cerebral perfusion pressure, while a control group of 175 patients underwent monitoring and management of cerebral perfusion pressure only. INTERVENTIONS: Routine neuroemergency procedures. MEASUREMENTS AND MAIN RESULTS: The two groups of patients were matched with regard to age, postresuscitation Glasgow Coma Scale scores, rates of acute surgical intracranial hematomas and brain swelling, pupillary abnormalities, early hypotensive events (before intensive care monitoring), as well as initial levels of intracranial pressure and cerebral perfusion pressure. Outcome at 6 months post injury was significantly better (p < .00005) in the 178 patients undergoing monitoring and management of cerebral extraction of oxygen along with cerebral perfusion pressure, than in the control group of 175 patients undergoing monitoring and management of cerebral perfusion pressure alone. CONCLUSION: In patients with severe acute brain trauma and intracranial hypertension associated with compromised cerebrospinal fluid spaces, monitoring and managing cerebral extraction of oxygen in conjunction with cerebral perfusion pressure result in better outcome than when cerebral perfusion pressure is managed alone.

Acute Disease↗
PubMed1998

Human cytotoxic T-lymphocyte repertoire to influenza A viruses.

The murine CD8(+) cytotoxic-T-lymphocyte (CTL) repertoire appears to be quite limited in response to influenza A viruses. The CTL responses to influenza A virus in humans were examined to determine if the CTL repertoire is also very limited. Bulk cultures revealed that a number of virus proteins were recognized in CTL assays. CTL lines were isolated from three donors for detailed study and found to be specific for epitopes on numerous influenza A viral proteins. Eight distinct CD8(+) CTL lines were isolated from donor 1. The proteins recognized by these cell lines included the nucleoprotein (NP), matrix protein (M1), nonstructural protein 1 (NS1), polymerases (PB1 and PB2), and hemagglutinin (HA). Two CD4(+) cell lines, one specific for neuraminidase (NA) and the other specific for M1, were also characterized. These CTL results were confirmed by precursor frequency analysis of peptide-specific gamma interferon-producing cells detected by ELISPOT. The epitopes recognized by 6 of these 10 cell lines have not been previously described; 8 of the 10 cell lines were cross-reactive to subtype H1N1, H2N2, and H3N2 viruses, 1 cell line was cross-reactive to subtypes H1N1 and H2N2, and 1 cell line was subtype H1N1 specific. A broad CTL repertoire was detected in the two other donors, and cell lines specific for the NP, NA, HA, M1, NS1, and M2 viral proteins were isolated. These findings indicate that the human memory CTL response to influenza A virus is broadly directed to epitopes on a wide variety of proteins, unlike the limited response observed following infection of mice.

Adult↗
PubMed1998

Assessment of human CD4+ and CD8+ T lymphocyte responses in experimental viral vaccine studies.

The traditional in vitro correlate of immunological protection is the induction by a vaccine of neutralizing antibodies against the virus. It was formerly assumed that protection induced by a vaccine was solely due to neutralizing antibodies. Neutralizing antibodies are potent in the prevention of certain diseases, but virus-specific CD4+ T helper cells aid in the proliferation of specific antigen-triggered B cells to make antibodies. CD8+ T cells are responsible for eliminating virus-infected cells during viral illness, and may act as a second line of defence by becoming activated and eliminating any cells that become infected despite the presence of neutralizing antibodies, for example because of a large challenge dose or antigenic variation at the antibody combining sites. We will briefly review our approaches for measuring human virus-specific CD4+ and CD8+ T cell responses to experimental vaccines. It is critically important to have sensitive, quality controlled assays, including positive controls. There are many potential variables in human T cell assays and pitfalls, which usually result in negative CTL results. Uninterpretable data are to be expected unless adequate preliminary testing has been done to establish sensitive, specific and controlled human antigen specific CD4+ and CD8+ T cell assays.

B-Lymphocytes↗
PubMed1997

Hantavirus pulmonary syndrome: CD8+ and CD4+ cytotoxic T lymphocytes to epitopes on Sin Nombre virus nucleocapsid protein isolated during acute illness.

In 1993 a number of cases of unexplained adult respiratory syndrome occurred in the southwestern United States. The illness was characterized by a prodrome of fever, myalgia, and other symptoms followed by the rapid onset of a capillary leak syndrome with hemoconcentration, thrombocytopenia, and pulmonary edema. Viral RNA sequences in the lungs identified a new member of the hantavirus genus, Sin Nombre virus (SNV), unique to North America. Pulmonary endothelial cells were heavily infected but were not necrotic. We speculated that this capillary leak syndrome was initiated by immune responses to the SNV-infected pulmonary endothelial cells. We isolated a CD8+ cytotoxic T lymphocyte (CTL) clone directly from the blood of a patient with the acute hantavirus pulmonary syndrome (HPS) which recognizes a SNV specific epitope on the virus nucleocapsid protein (aa 234-242) that is restricted by HLA C7 and produces IFN gamma but not IL-4. We identified a second CD8+ CTL epitope located within another site aa 131-139 on the nucleocapsid protein, which is HLA B35 restricted, and a CD4+ CTL epitope located on a third site on nucleocapsid protein aa 372-380 using lymphocytes obtained during HPS from another patient that were stimulated in vitro. Hantavirus specific CD8+ and CD4+ CTL may contribute to the immunopathology and capillary leak syndrome observed in the HPS.

Acute Disease↗
PubMed1997

Monosomy X as a recurring sole cytogenetic abnormality associated with myelodysplastic diseases.

Solitary loss of the X chromosome is associated with Turner syndrome and not hematological disorders. We describe five patients with non-constitutional loss of the X chromosome as the sole cytogenetic abnormality in their bone marrow. Three of the five patients had myelodysplastic syndrome (MDS), one case had AML M-6 with evidence suggestive of an evolving MDS, and the last patient had a dysplastic marrow. A review of the literature identified sporadic reports of an association of monosomy X and several hematologic disorders, as well as a few solid tumors. In this series of patients, monosomy X as a sole non-constitutional cytogenetic abnormality in bone marrow is associated with myelodysplastic diseases. In addition, fluorescence in situ hybridization analysis with an X centromere probe indicated that monosomy X was present in erythroid precursors, myeloblasts, promyelocytes, myelocytes, metamyelocytes, granulocytes, and monocytes, while mature lymphocytes presented with two copies of the X chromosome. The molecular cytogenetic evidence supports the diagnosis of a myelodysplastic disorder in these cases and documents the potential role of FISH in hematological disease.

Adult↗
PubMed1997

Prophylactic use of ivermectin against cattle myiasis caused by Cochliomyia hominivorax (Coquerel, 1858).

The prophylactic efficacy of ivermectin against navel or scrotal myiasis in calves was evaluated in eight trials in Argentina and Brazil. In two trials, calves were injected subcutaneously with ivermectin at a dosage of at least 200 microg kg(-1) within 24 h of birth. In the other six trials, two with two-month-old calves and four with four-month-old or older calves, all calves were treated with ivermectin at a dosage of at least 200 microg kg(-1) immediately after castration. In all trials, calves were maintained together on pasture and naturally exposed to Cochliomyia hominivorax. Navel and scrotal wounds were examined for myiasis daily for at least 14 days. Incidence of navel and scrotal myiasis was significantly lower (P < 0.01) in treated calves than in control calves.

Animals↗
PubMed1997

Immunologic responses to vaccinia vaccines administered by different parenteral routes.

To develop a less reactogenic but equally immunogenic vaccine, this study of 91 human volunteers compared the safety and immunogenic potency of a new, cell culture-derived vaccinia virus vaccine administered intradermally and intramuscularly with the licensed vaccinia vaccine administered by scarification. Cutaneous pox lesions developed in a higher proportion of scarification vaccinees. Scarification and intradermal vaccine recipients who developed cutaneous pox lesions had more local reactions but also achieved significantly higher cell-mediated and neutralizing antibody responses than those who did not develop pox lesions. Although less reactogenic, intradermal or intramuscular administration of vaccinia vaccine without the concomitant development of a cutaneous pox lesion induced lower immune responses.

Antibodies, Viral↗
PubMed1997

[Cerebral extraction of oxygen. A practical model and its clinical applications].

Cerebral extraction of oxygen (CEO2) represents a practical physiologic measure, with multiple clinical applications. This variable is calculated as the arterio-jugular difference in oxyhemoglobin saturation (from arterial blood and from the jugular bulb). Because it involves global measurements, the CEO2 does not allow detection of regional abnormalities in cerebral hemometabolism. However, when dealing with predominantly (not exclusively) global changes, the CEO2 provides accurate information on the balance (or coupling) between cerebral consumption of oxygen and cerebral blood flow.

Adult↗
PubMed1997

Cloning, expression and purification of recombinant streptokinase: partial characterization of the protein expressed in Escherichia coli.

We cloned the streptokinase (STK) gene of Streptococcus equisimilis in an expression vector of Escherichia coli to overexpress the profibrinolytic protein under the control of a tac promoter. Almost all the recombinant STK was exported to the periplasmic space and recovered after gentle lysozyme digestion of induced cells. The periplasmic fraction was chromatographed on DEAE Sepharose followed by chromatography on phenyl-agarose. Active proteins eluted between 4.5 and 0% ammonium sulfate, when a linear gradient was applied. Three major STK derivatives of 47.5 kDa, 45 kDa and 32 kDa were detected by Western blot analysis with a polyclonal antibody. The 32-kDa protein formed a complex with human plasminogen but did not exhibit Glu-plasminogen activator activity, as revealed by a zymographic assay, whereas the 45-kDa protein showed a K(m) = 0.70 microM and kcat = 0.82 s-1, when assayed with a chromogen-coupled substrate. These results suggest that these proteins are putative fragments of STK, possibly derived from partial degradation during the export pathway or the purification steps. The 47.5-kDa band corresponded to the native STK, as revealed by peptide sequencing.

Cloning, Molecular↗