Side population cells in highly enriched CD34-positive cells from peripheral blood progenitor cells identify an immature subtype of hematopoietic progenitor cells but do not predict time to engraftment in patients treated with high-dose therapy

Objective: A Hoechst 33342 dye efflux assay can be used to define a population of immature hematopoietic progenitor cells (HPC) that are called side population (SP) cells. Previously, SP cells examined from bone marrow (BM) and peripheral blood progenitor cells (PBPC) were found to be predominantly CD34 negative. Methods and results: In this study, we show that the level of CD34+ cells within the SP fraction increases from 2% in BM to 15% in mobilized PBPC. Furthermore, SP cells are found in highly enriched CD34+ cells from both BM and PBPC, and these cells define an immature phenotype of HPC. We also observed a higher level of CD133+ cells within the SPCD34+ cell population. Moreover, the frequency of long‐term culture‐initiating cells (LTC‐IC) was markedly increased in SPCD34+ cells. To further investigate whether variations in the level of SP cells in the CD34+ cell fraction influenced short‐term engraftment, we studied 20 patients with Hodgkin lymphoma that were autotransplanted with highly enriched CD34+ cells from PB. The percentage of SP cells in the PBCD34+ cell fraction was highly variable, ranging from 0.3 to 22%. No correlation was found between the content of SP cells in the autotransplanted CD34+ cells and time to short‐term engraftment. Conclusion: SPCD34+ cells in PBPC define an immature phenotype of HPC with increased numbers of LTC‐IC, and they are more frequently found in PBPC than in BM. The number of SP cells does not predict time to engraftment.     

Ki67 and 4F2 antigen expression as well as DNA synthesis predict survival at relapse/tumour progression in low-grade B-cell lymphoma

Previous work has shown that parameters of cell activation studied on lymphoma biopsies can be used to discriminate between low-grade and high-grade non-Hodgkin's lymphomas and to predict prognosis in the low-grade malignancy group alone. We have now examined expression of several activation antigens and indicators of DNA synthesis in 29 patients with low-grade malignant B-cell lymphomas at the time of primary diagnosis and later at relapse and/or tumour progression. At both times, the level of 4F2 antigen expression examined by flow cytometry on cells in suspension as well as the number of Ki67 antigen-positive cells examined by immunohistochemistry were predictive of patient survival. DNA synthesis estimated by (3H-TdR) thymidine incorporation was of prognostic value at the second biopsy only. These parameters were more sensitive than histological demonstration of morphological transformation in secondary high-grade lymphomas in identifying high-risk patients at repeated biopsy. We propose that Ki67 or 4F2 expression or a marker of DNA synthesis (such as 3H-TdR incorporation or labelling index) should be evaluated when repeated biopsies are performed, in order to select patients for whom aggressive chemotherapy may be considered.     

BMP-6 inhibits growth of mature human B cells; induction of Smad phosphorylation and upregulation of Id1

Background

Follicular dendritic cells (FDCs) play a central role in controlling B-cell response maturation, isotype switching and the maintenance of B-cell memory. These functions are based on prolonged preservation of antigen and its presentation in its native form by FDCs. However, when entrapping entire pathogens, FDCs can turn into dangerous long-term reservoirs that may preserve viruses or prions in highly infectious form. Despite various efforts, the ontogeny of FDCs has remained elusive. They have been proposed to derive either from bone marrow stromal cells, myeloid cells or local mesenchymal precursors. Still, differentiating FDCs from their precursors in vitro may allow addressing many unsolved issues associated with the (patho-) biology of these important antigen-presenting cells. The aim of our study was to demonstrate that FDC-like cells can be deduced from monocytes, and to develop a protocol in order to quantitatively generate them in vitro.

Results

Employing highly purified human monocytes as a starter population, low concentrations of Il-4 (25 U/ml) and GM-CSF (3 U/ml) in combination with Dexamethasone (Dex) (0.5 μM) in serum-free medium trigger the differentiation into FDC-like cells. After transient de-novo membrane expression of alkaline phosphatase (AP), such cells highly up-regulate surface expression of complement receptor I (CD35). Co-expression of CD68 confirms the monocytic origin of both, AP^pos and CD35^pos cells. The common leukocyte antigen CD45 is strongly down-regulated. Successive stimulation with TNF-α up-regulates adhesion molecules ICAM-1 (CD54) and VCAM (CD106). Importantly, both, AP^pos as well as AP^neg FDC-like cells, heterotypically cluster with and emperipolese B cells and exhibit the FDC characteristic ability to entrap functionally preserved antigen for prolonged times. Identical characteristics are found in monocytes which were highly expanded in vitro by higher doses of GM-CSF (25 U/ml) in the absence of Dex and Il-4 before employing the above differentiation cocktail.

Conclusion

In this work we provide evidence that FDC-like cells can be derived from monocytes in vitro. Monocyte-derived FDC-like cells quantitatively produced offer a broad utility covering basic research as well as clinical application.     

Vecuronium and danger of vagal induced cardiac arrest during laparotomy: A case report and literature review

Key words  vecuronium - bradycardia - cardiac arrest     

Action of opioid agonist-antagonist drugs on the pupil and nociceptive responses in mice

Opioid derivatives with mixed agonist-antagonist activities are becoming increasingly more popular in analgesia. We tested the mydriatic and analgesic activity of morphine in mice in comparison with similar effects of three agonist-antagonist agents: buprenorphine, butorphanol and nalbuphine. We also examined the antagonistic action of these three drugs by evaluating the analgesia and mydriasis in animals pretreated with morphine.The analgesic effect was assayed using the hot plate method while the pupillary responses were measured with a binocular operating microscope.Morphine produced dose-dependent mydriasis and analgesia in mice. The morphine-type agent buprenorphine and two nalorphine-type agonist-antagonists, butorphanol and nalbuphine, caused agonistic mydriatic and analgesic effects, usually less effective then morphine. Buprenorphine proved to have higher agonist activity than butorphanol and nalbuphine. The difference between butorphanol and nalbuphine was not statistically significant.A correlation between the mydriatic and the analgesic activity, known to exist among opioid derivatives with agonist activity only, was also demonstrated in the three investigated agonist-antagonist agents.Morphine-induced mydriasis and analgesia were reversed by all three agonist-antagonist drugs, but buprenorphine is a significantly weak antagonist in comparison with butorphanol and nalbuphine. An antagonistic property (antimydriatic and antianalgesic effects after pretreatment with morphine) of both nalorphine-type investigated drugs was not statistically significant, except for the antianalgesic effect of nalbuphine in doses 1 and 3mg·kg^–1, which was higher in comparison with butorphanol.(Stav A, Rabinowitz R, Korczyn AD: Action of opioid agonist-antagonist drugs on the pupil and nociceptive responses in mice. J Anesth 6: 439–445, 1992)     

Is the brain water channel aquaporin‐4 a pathogenetic factor in idiopathic intracranial hypertension? Results from a combined clinical and genetic study in a Norwegian cohort

Purpose: Idiopathic intracranial hypertension (IIH) is a condition of increased intracranial pressure of unknown aetiology. Patients with IIH usually suffer from headache and visual disturbances. High intracranial pressure despite normal ventricle size and negative MRI indicate perturbed water flux across cellular membranes, which is provided by the brain water channel aquaporin‐4 (AQP4). IIH could be associated with malfunctioning intracerebral water homeostasis and cerebrospinal fluid (CSF) reabsorption based on functional or regulatory alterations of AQP4. Methods: Clinical data, blood and CSF samples were collected from 28 patients with IIH. Clinical characteristics were assessed, and a genetic association study was performed by sequencing the AQP4 gene on chromosome 18. Genetic data were compared with 52 healthy controls and matched by age, sex and ethnicity. Chi‐square test and linear discriminant analysis (LDA) were used in the search of a genotype–phenotype association. Results: While the majority of patients responded to medical treatment, four required shunt application. All, except one, had a good visual outcome. The 24 AQP4 gene SNPs showed no association with IIH. Full cross‐validation of the LDA modelling resulted in only 55.1% correct classification of the cases and controls, with a corresponding estimated p‐value 0.37. Conclusions: Our genetic case–control study did not indicate an association between AQP4 gene variants and IIH. However, the theory of an etiopathogenic link between IIH and AQP4 is tempting, and discussed in this article. Association studies with large sample size are difficult to perform owing is the rarity of the condition.     

Review of the evidence related to older adult community mobility and driver licensure policies.

A systematic review of literature related to the effects of drivers' license policies and community mobility programs on older adult participation was completed as a part of the American Occupational Therapy Association's Evidence-Based Literature Review Project. The results revealed that states can achieve a goal of reducing traffic crashes, traffic violations, and traffic-related fatalities through relicensing policies requiring in-person renewal and vision testing as well as driving restrictions. Evidence suggests that failure to consider the continued community mobility needs of older adults can result in increasing fatalities among senior citizens using other modes of community mobility. This evidence-based review is informative for occupational therapy practice; however, it does not reveal the association between policy and community mobility interventions and improved engagement and participation. There is a role for occupational therapy in policy development and community mobility programming to promote engagement in occupation to support participation.     

Suicidal ideation and suicide attempts in a nationwide sample of operational Norwegian ambulance personnel

This is the first paper on suicidal ideation and attempts among ambulance personnel. This study aimed to investigate levels of suicidal ideation and suicide attempts among ambulance personnel, and to identify important correlates and the factors to which ambulance personnel attribute their serious suicidal ideation. A comprehensive nationwide questionnaire survey of 1,180 operational ambulance personnel was conducted. Measurements included: Paykel's Suicidal Feelings in the General Population questionnaire, the Hospital Anxiety and Depression scale, the Subjective Health Complaints Questionnaire, the Maslach Burnout Inventory, the Job Satisfaction Scale, the Basic Character Inventory, and the Rosenberg Self-Esteem Scale. Lifetime prevalence ranged from 28% for feelings that life was not worth living to 10.4% for seriously considered suicide and 3.1% for a suicide attempt. Serious suicidal ideation was independently associated with job-related emotional exhaustion (feelings of being overextended and depleted of resources) (OR 1.5, 95% CI 1.1-2.0) and bullying at work (OR 1.7, 95% CI 1.02-2.7), younger age, not married/cohabitant, depression symptoms, low self-esteem and the personality trait reality weakness. In general, suicidal thoughts were hardly attributable to working conditions, since only 1.8% of ambulance personnel attributed suicidal ideation to work problems alone. In conclusion, ambulance personnel reported a moderate level of suicidal ideation and suicide attempts. Although serious suicidal ideation was rarely attributed to working conditions in general, this study suggests that job-related factors like emotional exhaustion and bullying may be of importance.     

CT in children with abdominal cancer: should we routinely include the pelvis?

Background: It has been suggested that the pelvis should not be habitually included on abdominal CT examinations, but the potential benefit of such a practice in childhood abdominal malignancies is unknown. Objective: To estimate the yield and potential diagnostic benefit of abnormal findings on CT of the pelvis in children with malignant primary tumours in the upper abdomen. Materials and methods: From a paediatric tertiary referral hospital we retrospectively included patients having abdominal CT for primary upper abdominal tumours (1997–2004), the scan range routinely including the pelvis. We reviewed and tabulated any pelvic abnormality, and calculated group proportions with 95% confidence intervals. Results: We identified 230 children (2 days to 17 years old, median 2.9 years). Six (2.6%; 95% CI 0.5–4.7%) had abnormalities in the pelvis that would not have affected clinical management. Four (1.7%; 95% CI 0.1–3.4%) had findings that might have influenced staging, but only one was not detected by other modalities within 1 week of the CT. Conclusions: Our data suggest that diagnostically significant findings in the pelvis are rare; consequently, the habitual inclusion of the pelvis on abdominal CT for primary malignant tumours in the abdomen is not justified.