Differential effects of CD4+ and CD8+ cells on lymphocyte development from human cord blood cells in murine fetal thymus explants.

The possibility that mature lymphocytes play a role in the regulation of human T cell development was studied in the experimental model of fetal thymus organ cultures (FTOC), by reconstituting lymphocyte-depleted murine fetal thymus (FT) lobe with cells isolated from human umbilical cord blood (CB). Cultures were incubated with human cytokines (IL-7, FLT-3 ligand and Steel Factor), or remained untreated. When CD4+, or CD8+ CB cells, were co-cultured with FT explants, they expanded and maintained their original phenotypic markers, with no significant effect of the cytokines. Cultures of human hematopoietic stem cells (CD34+) gave rise to CD4+CD8- cells, which were mainly CD3-, with no indication of further intermediate developmental stages. However, a limited number of CD4+CD8+ (double positive [DP]) cells were detected when the CD34- cells were co-cultured with CD4+ cells from the same CB samples. In contrast, FT with unseparated CB cells resulted in the different CD4/CD8 subsets, and their numbers increased in the presence of cytokines. The appearance of DP cells depended on the presence of either CD4+ or CD8+ cells in the cultured CB samples. Hence, DP cells were not detected when the CB was depleted of CD4+ and CD8- cells ("depCB") before culture, and they appeared when depCB were co-cultured with either CD4+ or CD8+ cells. In contrast, CD4+ cells inhibited the development of CD8+CD3+ cells, and this was most pronounced in the absence of the cytokines. There was no symmetrical down-regulatory effect of CD8+ cells on the development of CD4+CD3+ cells. Addition of IL-15 to the cytokine mixture led to an increased proportion of CD56+ cells in cultures of CD34+ cells. The presence of CD4+, and not CD8+ cells, interfered with this process. Our results thus imply differential effects of CD4+ and CD8+ cells on thymocytopoiesis.

3H-estradiol, 3H-testosterone and 3H-dihydrotestosterone localization in the brain of the lizard Anolis carolinensis: an autoradiographic study.

The presence and the neuroanatomical topography of sex hormone concentrating cells in the brain of the American chameleon, Anolis carolinensis have been demonstrated by these experiments. After 3H-estradiol administration large numbers of hormone concentrating cells were found in the amygdala, septum, medial preoptic area, anterior hypothalamic area, the ventromedial and periventricular nuclei of the hypothalamus, and anterior pituitary. In addition, labelled cells were found in the torus semicircularis, in and around the nucleus isthmus pars parvocellularis. A small number of labelled cells could also be found in the rostral pallium, motor nucleus of the fifth cranial nerve, the raphé nuclei, and the spinal cord. After 3H-testosterone or 3H-dihydrotestosterone administration the neuroanatomical pattern was very similar to that found after 3H-estradiol; however, fewer labelled cells were seen after the androgens were given. Two exceptions to the similarity of pattern were in a caudal part of the pallium and in the mesencephalic tegmental area. Hormone-concentrating cells were found after 3H-testosterone or 3H-dihydrotestosterone administration, while labelled cells in these two areas after 3H-estradiol administration were extremely rare. The pattern of hormone-concentrating cells was the same in male and female brains, for each of the hormones. The preoptic area, hypothalamus, and anterior pituitary have been demonstrated in reptiles to be involved in neuroendocrine regulation and in the control of sex behaviors. The presence and neuroanatomical pattern of sex steroid binding cells in the brains of a wide variety of vertebrates have been documented. Large numbers of hormone-concentrating cells were found in all of these species in the medial preoptic area, tuberal hypothalamus, specific limbic structures, the mesencephalon deep to the tectum, and the anterior pituitary. Most hormone-concentrating cells in the brain of A. carolinensis were found in these same brain regions, thus indicating a vertebrate-wide stable core of hormone-concentrating cells in neuroanatomically defined regions.

Amplification of immunological functions by subcutaneous injection of intermediate-high dose interleukin-2 for 2 years after autologous stem cell transplantation in children with stage IV neuroblastoma.

BACKGROUND: Immunotherapy given post-autologous stem cell transplantation may eliminate residual tumor cells escaping the conditioning protocol. METHODS: Five children suffering from stage IV neuroblastoma were treated by recombinant interleukin-2 (IL-2) post-autologous peripheral blood stem cell transplantation. The patients' peripheral mononuclear cells were monitored for CD3+ and CD56+ levels, their proliferative response and killing of various cell lines targets. RESULTS: An increase in the level of total lymphocytes, mainly due to expansion of T cells, and enhanced proliferative response to phytohemaglutinin were observed. Elevated cytotoxicity against K562 and neuroblastoma target cells was detected in four patients and against K562 targets in one patient. Toxicity included mild thrombocytopenia, and fever in four patients and mild to moderate encephalopathy which necessitated withdrawing one patient from the protocol. Three of five patients studied are alive today, one of them whose IL-2 was stopped, is in relapse. Two patients have died. CONCLUSIONS: Immunotherapy with s.c. intermediate-high dose IL-2 is feasible and results in expansion of T cells and in stimulation of killing activity against several targets including in some cases, neuroblastoma tumor cells.

Effect of granulocyte depletion in a ventilated surfactant-depleted lung.

In a previous paper Cutz, Bryan et al. showed that in rabbits after repetitive lung lavage high-frequency oscillatory ventilation maintained excellent gas exchange and did not cause hyaline membrane formation (J. Appl. Physiol. 55: 131-138, 1983). In contrast, conventional mechanical ventilation had poor gas exchange and extensive hyaline membrane formation and we attributed these differences to mechanical barotrauma. However, we completely overlooked the large number of granulocytes in the damaged lung. To investigate this using the same model we have used mechanical ventilation on two groups of rabbits, one with normal granulocytes, the other depleted of granulocytes by pretreatment with nitrogen mustard. The nondepleted rabbits had poor gas exchange, a substantial protein leak into the lung and extensive hyaline membranes. The depleted animals had good gas exchange, a very small protein leak and no hyaline membranes. Repletion of granulocytes from donor rabbits lead to poor gas exchange and hyaline membrane formation. It is concluded that lung lavage causes prompt margination of granulocytes which become activated by the ongoing epithelial barotrauma of conventional ventilation.

Acute lymphoblastic leukemia with a unique translocation in an adolescent boy.

We report a case of an adolescent boy with acute lymphoblastic leukemia whose blasts had three chromosomal abnormalities: trisomy 8, a t(5;15), and an extra "marker" chromosome. The patient presented with huge hepatosplenomegaly and pancytopenia. The response to treatment (ALL BFM 90 protocol) was very rapid, and the patient is in complete remission 1 year after diagnosis.

Translocation (1;20)(q32;q13.3) in myelofibrosis following polycythemia vera.

We report a novel chromosomal translocation (1;20)(q32;q13.3) in a patient with myelofibrosis following polycythemia vera. This 73-year-old woman developed myelofibrosis 6 years after the initial diagnosis of polycythemia vera (PV). The course of PV was uneventful. Subsequent to the diagnosis, the patient was treated with phlebotomy and low doses of hydroxyurea for 4 years. No therapy was delivered during the remaining 2 years. A bone-marrow biopsy and a karyotype analysis performed because of evolving anemia demonstrated myelofibrosis and a chromosomal aberration-t(1;20)(q32;q13.3). Aberrations in chromosomes 1 and 20 have been reported in myeloproliferative disorders, but a t(1;20) translocation has not been reported. Because a karyotype analysis was not performed at the time PV was diagnosed, whether this translocation is linked to the primary disease (PV) or to the transition to myelofibrosis is not known.

Deletion of the long arm of chromosome 7 in myelomonocytic leukemia with bone marrow eosinophilia.

We report a 62-year-old man with acute myelomonocytic leukemia with bone marrow eosinophilia (M4Eo), and a deletion of the long arm of chromosome 7. The patient presented with pancytopenia, which shortly after evolved to overt leukemia. There was no response to the daunorubicin-cytosine arabinoside (Ara-C) regimen, and a remission achieved with amsacrine (AMSA)-Ara-C lasted only for a short time. On relapse, a peculiar skin rash accompanied the hematologic picture. While ANLL with chromosome 7 abnormalities usually carries adverse prognosis, patients with M4Eo (which is usually associated with chromosome 16 abnormalities) do better. The patient described here examplifies that M4Eo may be associated with del(7)(q22), and that it is the chromosomal abnormality rather than the type of leukemia that might determine the clinical outcome.

Autologous umbilical cord blood transfusion.

The purpose of this study was to examine some aspects of umbilical cord blood collection for autologous transfusion in premature infants. All 120 microbacterial cultures (aerobic and anaerobic) of cord blood samples as well as 30 cultures of mycoplasma were treated. Cord prothrombin fragment (F 1 + 2) concentrations were quantified at one and 10 minutes after clamping of the cord. F 1 + 2 concentrations assessed on 25 newborn infants were similar and no linear association with time of clamping could be drawn. This means that cord blood thrombosis is not activated for at least 10 minutes following clamping of the cord. As far as is known, the first newborn infant to benefit from this method of transfusion is reported here. The premature infant received two portions of autologous blood (on days 5 and 7). No untoward effects were noted. Blood, collected from the umbilical cord, is a safe source for autotransfusion, provided that bacteriological testing has been carried out.

In vivo gene amplification in non-cancerous cells: cholinesterase genes and oncogenes amplify in thrombocytopenia associated with lupus erythematosus.

The ACHE and BCHE genes, encoding the acetylcholine hydrolysing enzymes acetylcholinesterase (ACHE) and butyrylcholinesterase (BCHE), co-amplify with several oncogenes in leukemic patients with platelet deficiency (thrombocytopenia). This and other experiments implicated ACHE and BCHE in the development of bone marrow megakaryocytes, the progenitors of platelets. Therefore, we wished to find out whether cholinesterase gene amplification would also occur in non-cancerous platelet disorders and, if so, whether oncogenes would amplify in such cases as well. The autoimmune disease systemic lupus erythematosus (SLE) presents an appropriate model system for this issue, since patients with SLE may suffer from thrombocytopenia resistant to most treatment modalities. Here, we report a 40-80-fold amplification of genomic sequences from the ACHE and BCHE genes as well as the C-raf, V-sis and C-fes/fps oncogenes in peripheral blood cells from an SLE patient with severe thrombocytopenia. PvuII restriction analysis and DNA blot hybridization of the amplified ACHE and BCHE sequences demonstrated apparent aberrations in both genes, suggesting that malfunctioning of modified, partially amplified cholinesterase genes may be involved in the etiology of thrombocytopenia associated with SLE. These observations imply that cholinergic mechanisms regulate megakaryocytopoiesis, shed new light on the diverse hematologic findings characteristic of SLE, and may become valuable as diagnostic, treatment and prognostic tools in the follow-up of patients suffering from thrombocytopenia associated with SLE. Furthermore, these findings reinforce the notion that cholinesterase gene amplifications are causally related with platelet abnormalities in multiple hemopoietic disorders.

Carnitine-deficient myopathy as a presentation of tyrosinemia type I.

Carnitine deficiency secondary to renal Fanconi's tubulopathy has been described in only a few inborn errors of metabolism: cystinosis, galactosemia, and Fanconi-Bieckel syndrome. We report a 27-month-old infant who presented with a sudden change in gait owing to proximal muscle weakness. The laboratory evaluation showed carnitine deficiency associated with Fanconi's tubulopathy. Eventually, tyrosinemia type I was diagnosed. Carnitine deficiency can contribute to the clinical picture of hepatorenal tyrosinemia and should therefore be evaluated and treated.

Benign intracranial hypertension associated with budesonide treatment in children with Crohn's disease.

Oral budesonide in adult studies is a potent corticosteroid with decreased systemic bioavailability and an improved adverse effect profile in comparison with prednisone. It has recently been introduced for the treatment of inflammatory bowel disease in Europe, Canada, and Israel. Benign intracranial hypertension has rarely been associated with corticosteroid therapy but has not been reported in association with budesonide therapy. Three adolescents with Crohn's disease and poor nutritional status developed benign intracranial hypertension while receiving oral budesonide. All three patients had previously received multiple courses of prednisone during the course of their disease, without developing intracranial hypertension. Benign intracranial hypertension resolved after medication withdrawal and did not recur with subsequent use of prednisone. Evaluation for benign intracranial hypertension should be considered in patients with inflammatory bowel disease who develop headache while receiving oral budesonide. This side effect may be associated with poor nutritional status.

Research in physical therapy: philosophy, barriers to involvement, and use among California physical therapists.

This study was designed to 1) determine the importance of research in physical therapy to professional physical therapists; 2) assess the factors impeding their involvement in research; and 3) examine the extent to which therapists use published research. The study was conducted in two parts, a series of personal interviews and a questionnaire developed from the results of the interviews. The sample for the interviews was drawn from the population of Santa Clara County, California, and the questionnaires were mailed to a random distribution of 300 physical therapists throughout the State of California. Lack of time and funding as well as unfamiliarity with both the research process and the use of statistics were cited as the main barriers to research involvement. Collaboration with other health professionals involved in research and an apprenticeships were listed as desirable methods to gain additional research skills; however, these were believed to largely unavailable. A large percentage of therapists used research information published in professional journals other than PHYSICAL THERAPY.

Short note: the potential of umbilical cord blood to increase tissue oxygenation in adult respiratory distress syndrome.

Umbilical blood, consisting mainly of fetal haemoglobin, has an increased oxygen affinity. Adult respiratory distress syndrome may be caused by any acute, diffuse, infiltrative lung lesion of diverse aetiologies and is characterized by severe arterial hypoxia. Mechanical ventilation with high FIO2 and elevated pressures is used to improve tissue oxygenation in these patients. Nevertheless, adult respiratory distress syndrome may be fatal. Our hypothesis suggests that transfusion of umbilical cord blood to adult respiratory distress syndrome patients may facilitate oxygen transport by increasing oxygen binding in the erythrocytes passing through the damaged lungs. The local hypoxia and the accompanied acidosis in the periphery may accelerate the oxygen unload to the tissues, thus augmenting overall oxygen delivery. Studies with animals and humans show that left-shifted oxyhaemoglobin dissociated curve confers a degree of adaptation to low-oxygen tension ambient. Umbilical cord blood is available in every hospital, and there are no contraindications to its use.

High rate of asthma among immigrants.

Articles dealing with the epidemiological aspects of asthma were carefully reviewed in order to support or reject our clinical impression of increased rate of asthma among immigrants. Particular emphasis was put on data on very high or very low rates of asthma. The proposed theories to explain these differences were critically examined. The prevalence of asthma in China and in Africa is 1-2% and 0.5-5%, respectively. The prevalence of the disease in other indigenous populations ranges between 0.5% and 12%. On the other hand, asthma is much more frequently seen in Australia and in New Zealand (approximately 20-25%), where peoples' ancestors immigrated from distant areas. Statistical meta-analysis found a significant difference between the rates of asthma in the two groups of populations (P < 0.001). Immunoglobulin E levels of immigrants in Sweden are higher than those of native Swedes. Similarly, cord blood immunoglobulin E concentrations are more elevated in neonates whose mothers emigrated to Germany from Eastern countries than in those of native German mothers. There is an increased rate of IgE-mediated asthma among immigrant populations.

Gender differences in the reactogenicity of measles-mumps-rubella vaccine.

BACKGROUND: In trials comparing different formulations of measles vaccine, excess non-specific mortality occurred in female children who received high titer vaccine. These findings suggest a gender-specific effect of measles vaccine. OBJECTIVES: To determine whether gender differences exist in the rates of adverse reactions and morbidity in the month following immunization with measles-containing vaccine, and to evaluate whether there is a gender-specific association between the humoral immune response to measles vaccination and post-vaccination morbidity. METHODS: Parents completed questionnaires on the health status of 755 infants aged 15-20 months, during the month preceding and the month following the measles-mumps-rubella vaccination. Blood samples were tested for measles antibody titers in a subsample of 237 infants. RESULTS: After controlling background morbidity in the infants, the relative risk of fever and rash following vaccination was 2.35 in females and 1.36 in males. The geometric mean antibody titers against measles were similar in both sexes and there was no significant association between antibody titer and post-vaccination morbidity in either sex. CONCLUSIONS: Our findings demonstrate higher rates of adverse effects in females following vaccination with MMR vaccine, irrespective of the humoral response. This study emphasizes the need to consider possible gender differences when evaluating new vaccines.

[Neonatal hemochromatosis].

Neonatal hemochromatosis (NH) is a rare disorder characterized by signs of hepatic insufficiency. The rapidly progressive course terminates in death in-utero or in the early neonatal period. Histopathologically, there is increased tissue iron deposition in many organs, particularly the liver, pancreas, heart and endocrine glands, but the extrahepatic reticuloendothelial system is relatively unaffected. Cases of NH are distributed in a pattern consistent with an error of metabolism inherited as an autosomal recessive trait. We present a premature infant who died 44 days postpartum of severe hepatic failure. Postmortem examination established the diagnosis of NH.

Prohepcidin concentrations and erythroid progenitors in cord blood of appropriate versus small for gestational age neonates.

OBJECTIVE: Prohepcidin (Pro-Hep), synthesized in the liver, is the prohormone of hepcidin (Hep), which reduces iron absorption in the gut; its synthesis is enhanced by inflammation and is reduced during hypoxia. We aimed to study the hypothesis that infants born small for gestational age (SGA) have reduced cord blood concentrations of Pro-Hep. STUDY DESIGN: Cord blood was collected from 20 SGA (term and near term >35 week gestation) infants and 20 appropriate for gestational age (AGA) controls. We excluded infants exposed to maternal chronic diseases, smoking, diabetes, alcohol or drug use. Both groups had a 1 min Apgar score above or equal to 7 and had normal cord blood pH (above 7.25). ELISA was used to determine serum concentrations of Pro-Hep and erythropoietin (EPO). Circulating CD71(+)/CD45(-)/SSC(low) cells were measured by flow cytometry as an index of erythroid progenitors. RESULT: There were no significant differences between groups in terms of hemoglobin concentrations, and Pro-Hep. In contrast, EPO levels and circulating CD71(+)/CD45(-)/SSC(low) erythroid progenitors were significantly higher in the SGA group. These differences remained significant even after controlling for gestational age and gravidity. CONCLUSION: Contrary to EPO upregulation during intrauterine growth restriction (IUGR), and higher concentrations of circulating erythroid progenitors, Pro-Hep concentration is not affected by IUGR.