Agreements and uncertainties in autologous haematopoietic stem cell mobilization and collection. A Spanish consensus document.

Although many experts position statements on autologous stem cell mobilization have been published, there are some aspects that are still under discussion. A Spanish Hematologist expert group was summoned to settle on agreements and uncertainties on PBSCs mobilization, including factors not always considered; as apheresis and cytometry key factors that determine a successful PBSC collection. This document reviews critical factors that define poor mobilizer patients and the tools to better collect the desired stem cells for a successful autologous haematopoietic stem cell transplant.

Adoptive immunotherapy with antiviral T cells: Methods and results. / Inmunoterapia adoptiva con linfocitos antivirales: métodos y resultados.

Viral infections and reactivations are one of the main causes of morbidity and mortality in patients who undergo allogeneic haematopoietic progenitor cell transplantation. Adoptive immunotherapy with virus-specific Tcells (from donor to patient) has shown efficacy in the antiviral treatment of patients who have undergone transplantation and whose immune system has not yet been reconstituted. Currently, and according to the requirements of the corresponding agencies that regulate the production of these advanced personalised therapies, the production and application of these cell products are being optimised in such a way that they comply with good manufacturing practice standards and are safe and effective for treating patients. To facilitate their implementation, we need to understand the foundations of producing and using virus-specific Tcells. This study reviews the evolution of the methodology for producing antiviral Tcells and the studies that support their therapeutic efficacy. The study covers up to the current production platforms, whose commercialisation has begun in Spain. These platforms will help obtain virus-specific Tcells and chimeric antigen receptor Tcells, among others, in a completely automated manner and under good manufacturing practice conditions. The implementation of these new methodologies in the Spanish healthcare system will undoubtedly facilitate patients' access to a new repertoire of advanced therapies.

Genomic full-length sequence of two new HLA-C alleles, HLA-C*04:239 and HLA-C*05:137.

Two novel HLA-C alleles, C*04:239 and C*05:137, were characterized in Spanish individuals.

An active haemovigilance programme characterizing the safety profile of 7437 platelet transfusions prepared with amotosalen photochemical treatment.

BACKGROUND: An active haemovigilance programme was implemented to survey adverse events (AE) associated with transfusion of platelets photochemically treated with amotosalen and ultraviolet A (PCT-PLT). The results of 5106 transfusions have already been reported. Here we report the results of an additional 7437 PCT-PLT transfusions. METHODS: The focus of this ongoing haemovigilance programme is to document all AEs associated with PCT-PLT transfusion. Data collected for AEs include: time of event after starting transfusion, clinical descriptions, vital signs, results from radiographs and bacterial cultures, event severity (Grade 0-4) and causal relationship to PCT-PLT transfusion. RESULTS: One thousand four hundred patients (mean 60 years, range 1-96) received PCT-PLT transfusions. The majority of the patients (53.4%) had haematology-oncology diseases and required conventional chemotherapy (44.8%) or stem cell transplantation (8.6%). Sixty-eight PCT-PLT transfusions were associated with AE. Acute transfusion reactions (ATR), classified as an AE possibly related, probably related, or related to PCT-PLT transfusions were infrequent (n = 55, 55/7437 = 0.7%) and most were of Grade 1 severity. Thirty-nine patients (39/1400 = 2.8%) experienced one or more ATRs. The most frequently reported signs/symptoms were chills, fever, urticaria, dyspnoea, nausea and vomiting. Five AEs were considered severe (> or = Grade 2); however, no causal relationship to PCT-PLT transfusion was found. Repeated exposure to PCT-PLT did not increase the likelihood of an ATR. No cases of transfusion-related acute lung injury and no deaths due to PCT-PLT transfusions were reported. CONCLUSIONS: Routine transfusion of PCT-PLT is well-tolerated in a wide range of patients. ATRs related to PCT-PLT transfusion were infrequent and most were of mild severity.

Cidofovir treatment of human polyomavirus-associated acute haemorrhagic cystitis.

We report the case of an 18-year-old patient who received an allogeneic bone marrow transplant from an HLA-identical unrelated donor for a Ph+ acute lymphoblastic leukemia, in his third complete remission. Cyclophosphamide and busulfan were used as conditioning treatment. Acute graft-versus-host disease developed on day +9, and the response to adequate treatment (steroids) was favourable. On day +45 the patient developed an acute severe haemorhragic cystitis, and BK polyomavirus was demonstrated in urine samples using electron microscopy and polymerase chain reaction. Urinary symptoms did not improve in spite of palliative treatment, but a response was evident after 2 weeks of cidofovir treatment.

Allogeneic peripheral stem cell transplantation in a case of hereditary sideroblastic anaemia.

We report on a case of pyridoxine refractory hereditary sideroblastic anaemia (HSA) in a 19-year-old man who underwent peripheral blood stem cell transplantation (PBSCT) from his HLA-identical brother. By using short tandem repeat polymorphism, 100% donor cells were observed in peripheral blood on day +21; bone marrow showed mixed chimaerism from day +21 to day +221, when 100% cells of donor origin were observed. The patient developed extensive chronic graft-versus-host disease with favourable response to treatment. When the haemoglobin range was normal, a programme of phlebotomies reduced serum ferritin levels. Three years after transplantation, the patient has an ECOG rating of 0, with completely normal haemoglobin values (15 g/dl). To our knowledge, this is the first PBSCT reported in a case of hereditary sideroblastic anaemia.

Randomized study of aprotinin and DDAVP to reduce postoperative bleeding after cardiopulmonary bypass surgery.

BACKGROUND: Patients on cardiopulmonary bypass (CPB) have an increased susceptibility to postoperative bleeding. Previous reports using desmopressin acetate (DDAVP) for the prevention of postoperative bleeding have given contradictory results, whereas the protease inhibitor aprotinin has been shown to reduce blood loss after this type of surgery. This randomized study was performed to assess the efficacy of DDAVP versus aprotinin in the prevention of bleeding after CPB. METHODS AND RESULTS: One hundred nine of 122 eligible patients were randomized to four different groups: Group A (n = 28) received aprotinin starting with a bolus of 2 x 10(6) KIU followed by a continuous infusion of 0.5 x 10(6) KIU/h until the end of surgery; group B (n = 25) received of DDAVP 0.3 micrograms/kg i.v. on completion of CPB; group C (n = 28) received two doses of DDAVP, the first as in group B and an additional dose 6 hours after surgery; group D (n = 28) received no treatment. There was a marked reduction of postoperative blood loss either at 12 hours (P < .01) or 72 hours (P < .02) in the aprotinin group compared with all other groups, whereas no significant effect was observed in either of the two DDAVP regimens. A significant reduction in the amount of blood used was observed only in the aprotinin group (P < .01). Of the plasma fibrinolytic components assayed, there was a significant reduction of the fibrin degradation product generation in the aprotinin group (P < .001), whereas a significant systemic hyperfibrinolysis was observed in both DDAVP-treated groups and the control group. No side effects related to the study drugs were observed in any patient. CONCLUSIONS: Aprotinin inhibited fibrinolysis; this correlated with a significant reduction of postoperative blood loss and need for blood replacement after CPB. Neither one nor two doses of DDAVP had a beneficial effect. Aprotinin offers a better alternative than DDAVP in the prevention of bleeding after CPB.

[Does propofol have advantages over midazolam and isoflurane? Comparative study of 2 total intravenous anesthesia techniques using midazolam and propofol, versus balanced anesthesia with isoflurane]. / Tiene ventajas propofol sobre midazolam e isoflurano? Estudio comparativo de dos técnicas de anestesia total intravenosa con midazolam y propofol, y anestesia balanceada con isoflurano.

OBJECTIVES: To compare two techniques for total intravenous anesthesia (TIVA): midazolam-alfentanil-flumazenil and propofol-alfentanil, contrasting them with combined anesthesia (thiopental-isoflurane-alfentanil) and assessing the efficacy of flumazenil in continuous perfusion for preventing resedation in TIVA with midazolam. PATIENTS AND METHODS: The efficacy and clinical tolerance of the 3 anesthetic techniques with propofol, midazolam or isoflurane were studied in 63 patients undergoing elective breast, lumbar or gynecological surgery. Anesthetic induction was achieved with midazolam 0.3 mg/kg-1 (group M), propofol 2.5 mg/kg-1 (group P) or thiopental 3 mg/kg-1 (group I); all patients also received 50 micrograms/kg-1 alfentanil and vecuronium bromide 0.12 mg/kg-1/h-1. Maintenance was achieved with midazolam in perfusion at 0.12 mg/kg-1/h-1 (group M); propofol in perfusion at 7 mg/kg-1/h-1 and a pre-incision dose of 1.5 mg/kg-1 (group P); and isoflurane at 1.15% (group I). The 3 groups also received one pre-incision dose of alfentanil 25 micrograms/kg-1 and post-incision perfusion at 60 micrograms/kg-1/h-1. The infusion of alfentanil was changed by amounts of 20 micrograms/kg-1/h-1 in accordance with the patient's response to surgery. After surgery patients in group M received flumazenil 0.5 mg i.v. over 30 sec and a perfusion of flumazenil 0.5 mg over 60 min. Parameters indicating efficacy were: 1) total dose and timing of alfentanil; 2) number of instances of inadequate anesthesia; 3) peri-operative amnesia; 4) times of awakening and extubation after surgery, and 5) the number of patients in each group who required naloxone. Parameters indicating tolerance were: 1) hemodynamic variables; 2) the number of postoperative desaturations; 3) level of sedation, comprehension and motor coordination and orientation; 4) the "G/g detection" test and the memory recall test; 5) adverse side effects; 6) need for postoperative analgesia, and 7) evaluation of the anesthetic technique. RESULTS: The 3 techniques afforded effective control of hemodynamic response to intubation and surgical incision. Anesthetic maintenance was easy and safe with isoflurane and propofol. Higher doses of alfentanil, however, were needed with midazolam and we found a higher incidence of signs of superficial anesthesia. Reversion of midazolam with flumazenil 0.5 mg i.v. produced earlier awakening, although this was followed later by relapse into hypno-sedation that could not be prevented with a perfusion of flumazenil. Although recovery from anesthesia was slower with propofol than with isoflurane, we observed no differences in level of sedation, motor coordination and postoperative comprehension. Maintenance with isoflurane produced a higher incidence of adverse side effects such as tremors and nausea after surgery. CONCLUSIONS: None of the TIVA techniques proved superior in all the parameters studied during anesthetic maintenance when compared with balanced isoflurane-alfentanil, although the propofol-alfentanil combination was found to be superior to that of midazolam-alfentanil. After anesthesia, however, recovery was better with the association of propofol-alfentanil and adverse side effects were fewer. Flumazenil at the doses used was ineffective for preventing resedation due to midazolam.

[Comparison of the effects of isoflurane and alfentanyl on the mononuclear-phagocytic system]. / Comparación de los efectos del isoflurano y del alfentanilo sobre el sistema mononuclear-fagocítico.

BACKGROUND AIMS: The effects of two anesthetic agents, alfentanil and isoflurane, on the macrophage-monocytic system and the relation with the adrenocortical-hypophyseal axis were analyzed. MATERIAL AND METHODS: Thirty ASA I-II patients submitted to elective surgery were distributed into two groups: group I, analgesic anesthesia with alfentanil 100 micrograms/kg, a perfusion of 2-3 micrograms/kg/min and naloxone 0.2-0.4 mg upon termination of surgery; group II, inhalation anesthesia with isoflurane at 2-2.5% and O2/air = 1/1. HLA-DR membrane receptors and vimentin threads of the cytoskeleton were determined in basal conditions, at one hour and two hours of induction and upon termination of surgery by means of indirect immunofluorescence with the use of monoclonal antibodies. Phagocytosis of latex particles was also measured and ACTH and cortisol levels were obtained by radioimmunoassay. RESULTS: In group I (alfentanil) a decrease (p < 0.01) of all the monocyte parameters analyzed, which was more significant than that observed in group II (isoflurane), was observed. In group II only the expression of HLA-DR and the index of phagocytosis (p < 0.05) were statistically significant. Following the administration of naloxone a reversion of these parameters was observed. The maximum immune depression corresponded with the lowest values of ACTH and cortisol (group I) with surgical stress being discarded as the cause of this immunodepression. CONCLUSIONS: Alfentanil produces a monocytic depression which reverses following the administration of naloxone, thus discarding surgical stress as the cause of this immunodepression suggesting a mechanism mediated by opiate receptors. With regards to isoflurane, the functions of the mononuclear-phagocytic system were hardly altered in the patient studied.

[Depression of the mononuclear phagocyte system caused by high doses of narcotics]. / Depresión del sistema mononuclear-fagocítico provocada por altas dosis de morfínicos.

We evaluated in human monocytes the effect of high doses of alfentanyl on the expression of vimentin filaments, the phagocytic activity and the membrane display of HLA-DR molecules in the subjects undergoing surgery. The study was performed on 30 patients, ASAI-II. The patients received 100 mcg/kg i.v. of Alfentanil and the maintenance of anaesthesia was made with Alfentanil (2-3 mcg/kg/min.). The patients were randomized in two groups. The patients were ventilated with N2O:O2 (1:1) (Group I) or air: O2 (1:1) (Group II). After surgery, all patients of the Group II received Naloxone (0.2-0.4 mg). Central venous blood samples were obtained before induction, one and two hours after induction of anaesthesia and at the end of surgery. Separation of monocytes was performed according to Boyum technique. CD35 and HLA-DR molecules and vimentin filaments were studied by indirect immunofluorescence method using monoclonal antibodies. Percentage of positive cells were read with a cytofluorometer. The phagocytic function of monocytes was determined by ingestion of latex particles. Cortisol and ACTH plasma levels were determined by RIA. High doses of Alfentanyl depress phagocytic function and membrane display of CD35 and HLA-DR molecules in monocyte and induce marked changes in the organization of vimentin filaments in these cells in patients undergoing surgery. This monocytic depression was more marked in the patients ventilated with N2O. In our results there was uninhibition of ACTH and cortisol plasma levels responses to surgical stress by Alfentanil administration. Since the effects of Alfentanil were reversed by Naloxone, an opioid receptor mechanism seems to mediate these events.

Opioid peptides modulate the organization of vimentin filaments, phagocytic activity, and expression of surface molecules in monocytes.

It is theorized that intermediate filaments are important in the modulation of membrane activity and cell motility; however, their functions are unknown. The assembly and organization of these filaments are under hormonal regulation. We investigated in human monocytes the in vitro effects of Met-enkephalin, Leu-enkephalin, and beta-endorphin on the expression of immunoreactive cytoskeletal vimentin filaments. We simultaneously examined their effect on the phagocytosis of Candida albicans and on the membrane display of surface molecules. The three opioid peptides markedly reduced the expression of vimentin filaments, the phagocytic activity, and the display of HLA-DR molecules at concentrations of 10(-6), 10(-8), and 10(-10) M. On the other hand, the intravenous administration of fentanyl, a synthetic opiate agonist, to patients undergoing surgery induced similar changes in monocytes. In other experiments, 10(-8) M beta-endorphin also decreased the expression of CR3 but did not influence the display of CD13, a surface protein of unknown function. Expression of vimentin filaments correlated directly with the display of HLA-DR antigens and CR3 and with the phagocytic activity. The results of this paper indicate that opiates and opioids, neuropeptides known to be released during stress, can directly depress several monocyte functions. Furthermore, from these data it may be speculated that intermediate filaments may regulate the membrane expression of some surface molecules and the phagocytic process.

[A bupivacaine-morphine combination by intrathecal route: correlation between pain relief and postoperative neuroendocrine response]. / Asociación bupivacaína-morfina por vía intratecal: correlación entre alivio del dolor y respuesta neuroendocrina postoperatoria.

The purpose of the present study was to assess the repercussion of morphine injected in the intrathecal space on postoperative neuroendocrine response and the correlation with pain relief in the postop period. We studied 50 healthy patients (ASA I-II) submitted to orthopaedic surgery under general anaesthesia (N = 25) or spinal anaesthesia (N = 25). In the group under general anaesthesia we observed a hypersecretion of ADH, ACTH, cortisol and aldosterone during and after surgery. In the group un spinal anaesthesia, it was evident, on the contrary, a blockade of the neuroendocrine response during surgery, as well as an attenuation during postoperative period. Intraoperative and postoperative bleeding with spinal anaesthesia was significantly lower (p less than 0.01; p less than 0.05 respectively) than with general anaesthesia. Postoperative analgesia was excellent in group with spinal anaesthesia; the average duration of analgesia was 16.3 hours. We conclude that small intrathecal doses of morphine have beneficial effects and may be used usefulness in orthopaedic surgery.

The effect of labetalol on urinary prostacyclin secretion during surgery.

The effects of labetalol on the secretion of prostacyclin and plasma-renin activity (PRA) were evaluated, relative to a control group in 24 patients undergoing hip osteotomy. They were randomly assigned to two groups (G-I and G-II) with 12 patients each. Patients allocated to both groups received standard anaesthesia (thiopentone, pancuronium, fentanyl and nitrous oxide). Patients belonging to Group II were given labetalol at a dose of 0.8 mg kg-1. The stable metabolite of PGI2, 6-keto-PGF1 alpha was quantified from urine samples by radioimmunoassay (RIA). Cortisol, PRA and aldosterone were determined from blood samples. A significant increase in 6-keto-PGF1 alpha elimination was observed in G-I. Labetalol administration partially but significantly inhibited this increase. We believe that prostacyclin is involved not through the beta 1 but through the alpha 1 receptors in the secretion of renin.

The effects of epidural analgesia and conventional anaesthesia on renal excretion of PGE2 during orthopaedic surgery.

The effect of standard anaesthesia (G-I) and epidural analgesia (G-II) on urinary synthesis of PGE2 was compared in two groups of 12 patients each undergoing orthopaedic surgery. This study demonstrates a significant elevation of renal PGE2 in patients undergoing standard anaesthesia during surgery. In marked contrast, with epidural analgesia, the inhibition of renal secretion of PGE2 is apparent. These findings strongly suggest the important role of the sympathetic nervous system in the synthesis of PGE2.

[Changes in the renin-angiotensin-aldosterone system during open heart surgery. I. Factors involved in renin secretion]. / Modificaciones del sistema renina-angiotensina-aldosterona durante la cirugía a corazón abierto.

The endocrine, humoral and hemodynamic factors that affect renin secretion are studied during open-heart surgery in a group of 40 patients, 20 with mitral and 20 with aortic valve disease. During the incission period, the PRA (plasma renin activity) increases due to a sympatoadrenergic hyperactivity. These increase is noted also during bypass, and is closely related with the hypotension that extracorporeal circulation produces. It is also discussed the influence of the exclusion of pulmonary circulation, that decreases conversion of angiotensin I to II and catabolism of norepinephrine.

[Analgesics and neuroendocrine response in heart surgery: III. Metabolic aspects]. / Analgésicos y respuesta neuroendocrina en cirugía cardíaca: III. Aspectos metabólicos

A study done in 121 patients undergoing extracorporeal perfusion heart surgery for the correction of valvular cardiopathies. In a first stage, we evaluated, in 77 patients, the influence of different analgesics on renal function, acid-base balance, glucemia and lactic acid. Afterwards we completed our observation with analysis of insulin in 44 patients. We observed no important difference in glomerular filtration rate, urinary output per minute, acid-base equilibrium, or oxigenation, according to administration of the different analgesics. After the glucose overload which the extracorporeal perfusion imposes, we observed a better insulinic response and consequently a faster recuperation of normal levels of glucemia in patients who received Fentathienyl. The lowest response was obtained in patients anesthesized with Pentazocina. In view of these results we can affirm that analysis if the inter-relation between glucose and insulin is a good exponet for evaluation of the grade of analgesic protection, offered by a drug, against nociceptive stimuli triggered by surgery.

[Analgesics and neuroendocrinic response in cardiac surgery. I. Adrenal cortex response]. / Analgésicos y respuesta neuroendocrina en cirugía cardiaca. I.--Respuesta cortico-suprarrenal

In 91 patients who had undergone surgery under C.E.C. the influence of the analgesia on adrenal cortex system was studied. With the exception of the analgesic used, the anaesthesia was performed following the same guidelines in all of the operations. Concentrations of cortisol in plasma were maximum with Pentazocina, the weakest analgesic used by us, and minimum with Fentanest. In view of these results, we can affirm that, in the absence of alterations haemodynamic, metabolic and neurological, concentrations of cortisol in plasma are a true reflexion of the amount of analgesic protection that different drugs offer against noxins stimuli triggered by the surgical act. These findings guarantee the intimate relationship that exists between the nervous system and the endocrine system and they are further confirmed by results obtained with a recently acquired analgesic Fentathienil, which is more potent than Fentanyl.