Maintenance rituximab after autologous stem cell transplantation in patients with mantle cell lymphoma.

BACKGROUND: High-dose therapy and autologous stem cell transplantation (ASCT) improves outcomes for patients with mantle cell lymphoma (MCL), but relapse ultimately occurs in most patients. Recently presented interim results from a phase III prospective trial suggest maintenance rituximab (MR) after ASCT for MCL improves progression-free survival (PFS). The maturation of these data and any benefit of MR on overall survival (OS) remain to be defined. PATIENTS AND METHODS: In this retrospective study, we examined a cohort of consecutive patients with MCL that underwent ASCT for MCL at our center and evaluated their outcomes according to whether they received MR after ASCT (n = 50) or did not (n = 107). MR was treated as a time-dependent covariate to account for variation in timing of its initiation. RESULTS: MR was associated with an improved PFS [hazard ratio (HR) 0.44; confidence interval (CI) (0.24-0.80), P = 0.007] and overall survival (OS; HR 0.46; CI 0.23-0.93, P = 0.03) following a multivariate adjustment for confounding factors with a median follow-up of ∼5 years. Grade 4 neutropenia was increased (34% versus 18%, P = 0.04) in the MR group, but no effect on the rate of mortality unrelated to relapse was observed. CONCLUSIONS: These data support that MR after ASCT for MCL confers a benefit in PFS and additionally suggest it may improve OS. General application of this strategy will require confirmation of benefit in prospective randomized trials.

Role of glutamate in coupling between bilaterally paired circadian clocks in Bulla gouldiana.

A group of electrically coupled basal retinal neurons (BRN) in the eye of the marine snail Bulla gouldiana generate a circadian rhythm in the frequency of compound action potentials (CAPs). CAPs are conducted to the contralateral retina via the optic nerves and the cerebral commissures to synchronize the rhythms of both eyes. CAPs can induce an excitatory postsynaptic potential (EPSP) in the contralateral BRNs that can lead to action potential generation. The pathway and mechanism of this bilateral coupling signal have not been elucidated, but the evidence suggests monosynaptic connections between the populations of pacemaker cells in both retinae. The study was designed to further characterize the coupling signal and investigate the role of glutamate as a neurotransmitter in this pathway. We found evidence supporting our hypothesis that glutamate, previously identified in BRNs by an immunocytological study, is involved in bilateral coupling. First, a combination of extracellular and intracellular electrophysiological recordings revealed that both electrically and optically evoked CAPs generate excitatory synaptic potentials and action potentials in contralateral BRNs. Application of glutamate also led to increased neuronal activity of individual BRNs both in the intact retina as well when isolated in cell culture. Lastly, glutamate-induced inward currents were characterized in cultured BRNs using perforated-patch recordings. The reversal potential was close to 0 mV, and the currents were sensitive to N-methyl-d-aspartic acid (NMDA) and non-NMDA antagonists. NMDA and AMPA, as well as aspartate, also induced distinct inward currents in BRNs. We conclude that glutamate can be used by BRNs as a transmitter to influence electrical activity in the contralateral pacemaker population. We propose that glutamate is required for synchronizing of the bilaterally paired retinal clocks producing a unified circadian timing signal.

Amine and amino acid transmitters in the eye of the mollusc Bulla gouldiana: an immunocytochemical study.

We identified putative transmitters of the photoreceptors and circadian pacemaker neurons and found candidates for efferent control in the eye of the marine mollusc Bulla gouldiana. Established antisera against octopamine, dopamine, serotonin, histamine, glutamate, gamma-aminobutyric acid (GABA), and taurine were used, and central ganglia were processed in parallel to evaluate general staining quality. Photoreceptors and circadian pacemaker cells both expressed immunoreactivity for glutamate and taurine. The eye and its sheath were devoid of GABA-like immunoreactive material, and none of the antisera directed against biogenic amines labelled cells or processes in the nervous tissue of the eye. However, dopamine and octopamine antisera stained large spherical granules (diameter 2-3 microm) contained in granular cells that are located in the connective tissue encapsulating the eye and the optic nerve. The serotonin antiserum revealed a sparse distribution of varicose axon fibers in the optic nerve and eye sheath. No histamine-immunoreactive processes were revealed in the eye. The functional significance of these findings for the molluscan eye and its circadian clock is discussed.

The fight and flight responses of crickets depleted of biogenic amines.

Aggressive and escape behaviors were analysed in crickets (Orthoptera) treated with either reserpine, a nonspecific depleter of biogenic amines, or the synthesis inhibitors alpha-methyltryptophan (AMTP) and alpha-methyl-p-tyrosine (AMT) to specifically deplete serotonin, respectively dopamine and octopamine. Standard immunocytochemical techniques were used to verify depletion from central nervous tissue, and determine the effective dosages. Reserpinized crickets became exceedingly lethargic and had severely depressed escape responses. However, they were still able to express all the major elements of the escalating sequences of stereotype motor performances that typifies normal aggressive behavior in the cricket. AMT and AMTP treatment had opposing influences on escape behavior, being enhanced by serotonin depletion, but depressed by dopamine/octopamine depletion. AMTP-induced serotonin depletion had no influence on aggressive or submissive behaviors. AMT-treated crickets could normally only be brought to fight by coaxing. Though capable of expressing aggressive behavior per se, agonistic encounters between AMT-treated crickets were shorter, and rarely involved actual physical interactions. Hence, although amines seem to have similar actions on escape behavior in insects and crustaceans, the aminergic control of aggression seems to be fundamentally different in these arthropods groups. We conclude that amines are not in principle required for the initiation and operation of the motor circuits underlying aggression in the cricket. However, octopamine and/or dopamine seem necessary for establishing a level of excitability sufficient for aggressive behavior to become overt in response to appropriate natural releasing stimuli.

A randomized British National Lymphoma Investigation trial of CHOP vs. a weekly multi-agent regimen (PACEBOM) in patients with histologically aggressive non-Hodgkin's lymphoma.

BACKGROUND: Between 1987 and 1991, the British National Lymphoma Investigation randomized 459 patients with non-Hodgkin's lymphoma with a large-cell component to either CHOP or the PACEBOM regimen. PATIENTS AND METHODS: Four hundred fifty-nine eligible patients were included in this trial, four hundred one with diffuse large-cell lymphoma and fifty-eight with diffuse mixed-cell lymphoma according to the Working Formulation. Two hundred twenty-six patients were randomized to receive CHOP and two hundred thirty-three to receive PACEBOM. The two arms of the trial were well balanced for all potential prognostic factors. RESULTS: The complete remission rate with PACEBOM was 64% and with CHOP 57% (NS). At eight years, the actuarial cause specific survival (CSS) in the PACEBOM arm is 59% compared to 49% in the CHOP arm (P = 0.09). Patients < 50 years of age fared significantly better in the PACEBOM arm both for CSS and overall survival (P = 0.002) and the CSS was also significantly improved in stage IV disease (P = 0.02). CONCLUSIONS: The mature data from this trial suggest that an etoposide-containing multi-agent weekly regimen can be superior to CHOP.

Colocalisation of taurine- with transmitter-immunoreactivities in the nervous system of the migratory locust.

The expression of taurine immunoreactivity (TAU-IR) by neurones immunoreactive for octopamine (OA-IR), gamma-aminobutyric acid (GABA-IR), and the C-terminal peptide sequence arginine-phenylalanine (RFamide-IR) was investigated in the migratory locust (Locusta migratoria). TAU-IR is colocalised with OA-IR in the dorsal unpaired median neurones, which are efferent neuroparacrine cells. TAU-IR is not, however, expressed by OA-IR interneurones in the thoracic ganglia and brain. The only other TAU-IR somata found with peripheral axons are the medial neurosecretory cells in abdominal ganglia that project to the neurohaemal organs. These cells exhibit RFamide-IR. The majority of TAU-IR somata in the thoracic abdominal nervous system exhibit GABA-IR. These cells correspond to populations of identified local and intersegmentally projecting inhibitory interneurones. TAU-IR is not, however, exhibited by the well-known GABAergic common inhibitor neurones, which have peripherally projecting axons. This differential distribution of TAU-IR in basically two, functionally different, neuronal subsets (efferent neurosecretory and neuroparacrine cells, inhibitory interneurones) conforms with the concept of taurine acing as a depressive agent to limit excitation during stressful conditions.

A randomised comparison of a third-generation regimen (PACEBOM) with a standard regimen (CHOP) in patients with histologically aggressive non-Hodgkin's lymphoma: a British National Lymphoma Investigation report.

A combination of cyclophosphamide, doxorubicin, vincristine and prednisolone (CHOP) has been a standard therapy for histologically aggressive non-Hodgkin's lymphomas for over 20 years, but several newer regimens, referred to as second or third generation, have been reported to give improved results in single-centre studies. Positive evidence from randomised trials has been lacking, and the British National Lymphoma Investigation therefore commenced a randomised comparison of CHOP vs a third-generation regimen, PACEBOM, in November 1987. A total of 459 eligible patients were entered into the trial: 226 in the CHOP arm and 233 in the PACEBOM arm. Overall, there was no significant difference in outcome between the two arms of the trial. In patients with stage IV disease there was an apparent improvement in survival for those treated with PACEBOM, but considerable caution must be exercised with such subgroup analysis.

A new method for double immunolabelling with primary antibodies from identical species.

There are several double immunolabelling methods but each has its drawbacks. More often than not, antibodies with the required specificities are available in only one species and their use normally produces false labels due to cross-reactivity. We describe a new and reliable technique for staining with primary antibodies from the same species, that can even be employed on tissues of the donor species. The protocol avoids cross-reactivities without loss in sensitivity, uses commercially available reagents and takes advantage of enzymatic detection, although it can be adapted for fluorescent labelling. Briefly, tissue is incubated with one primary antibody, followed by a peroxidase-coupled secondary antibody which is detected using amino ethyl carbazol to give a red reaction product. Meanwhile, the next primary antibody is coupled in vitro to a biotinylated secondary antibody and excess binding sites quenched with normal immune serum from the same species as the primary antibody. This complex is applied to tissue and detected by the avidin-biotin/alkaline phosphatase technique using naphthol-AS-MX-phosphate/Fast Blue BB to produce a blue label. In addition to extensive controls, the reliability and broad applicability of this method has been confirmed in (1) murine skin cryostat sections to co-visualize antigen-presenting cells (MHC class II-immunoreactive; "-ir') with either antigen detecting T lymphocytes (CD4-ir) or Langerhans cells (NLDC-145-ir) and (2) locust (Insecta) abdominal ganglion paraffin sections, where it is known that immunoreactivities for octopamine and a FMRFamide-related peptide are colocalized in only one, uniquely identifiable neuron.

Localization of octopaminergic neurones in insects.

This paper reviews data on the localization of octopaminergic neurons revealed by immunocytochemistry in insects, primarily the locusts Schistocerca gregaria and Locusta migratoria, cricket Gryllus bimaculatus, and cockroach Periplaneta americana. Supporting evidence for their octopaminergic nature is mentioned where available. In orthopteran ventral ganglia, the major classes of octopamine-like immunoreactive (-LI) neurones include: (1) efferent dorsal and ventral unpaired median (DUM, VUM) neurones; (2) several intersegmentally projecting DUM interneurones in the suboesophageal ganglion; other DUM interneurones are probably GABAergic; (3) a pair of anterior median cells in the prothoracic ganglion; (4) a single pair of ventral cells in most thoracic and some other ganglia; these appear to be plurisegmentally projecting interneurones. Eight categories of octopamine-LI neurones occur in the orthopteran brain. The basic projections of three types are described here: one class project to the optic lobes to form wide field projections. Another type descends to cross into the tritocerebral commissure and may invade the contralateral brain hemisphere. A further class is the median neurosecretory cells with axons in the nervi corpori cardiaci I. Available data for the honey bee Apis mellifera and moth Manduca sexta indicate that the octopamine-LI cell types found in orthopterans also occur in holometabolous insects. Immunocytochemical evidence suggests that some octopaminergic DUM cells contain an FMRFamide-related peptide and the amino acid taurine as putative cotransmitters.

Colocalization of octopamine and FMRFamide related peptide in identified heart projecting (DUM) neurones in the locust revealed by immunocytochemistry.

Immunocytochemical techniques are employed to reveal colocalization of octopamine with FMRFamide related peptide in the locust ventral nervous system. In each unfused pregenital abdominal ganglia (A4-A6) there are 3 octopamine-like immunoreactive neurones. By combining intracellular Lucifer yellow staining with subsequent immunocytochemistry these are individually identified as the efferent dorsal unpaired median (DUM) neurones DUM-1 and DUM-2, which innervate abdominal tergal and respectively sternal skeletal muscles, and DUM heart-1, an FMRFamide-like immunoreactive neurone which projects to the heart and associated alary muscles. Colocalization of octopamine- and FMRFamide-like immunoreactivity in DUM heart-1 is verified by alternate staining of consecutive sections. With respect to locust ventral ganglia, this investigation shows that colocalization of octopamine with an FMRFamide related peptide is restricted to a single DUM cell occurring in each abdominal ganglion 2-7, which most likely corresponds to segmental homologues of DUM heart-1.

Evidence for octopaminergic nature of peripherally projecting DUM-cells, but not DUM-interneurons in locusts.

In locusts, a median neuroblast in each segmental ganglion gives rise to numerous unpaired progeny--the well known peripherally projecting dorsal-, occasionally ventral-, unpaired median (DUM-, resp. VUM-) neurones together with the lesser known DUM-interneurones 12. We examine the reputed octopaminergic nature of this nerve cell lineage using an anti-octopamine serum recently developed by M. Eckert and J. Rapus 7. This antiserum labels in each segmental ganglion numerous midline neurones, identifiable as DUM- and VUM-cells by their some sizes and positions, projections in DUM-tracts and characteristic T-junctions with bilaterally projecting axons. All octopamine immunoreactive DUM-, and VUM-neurones appear to project to peripheral nerves; their numbers correspond to the number of peripherally projecting DUM- and VUM-neurones identified so far in the examined ganglia. Presumptive DUM-interneurones, i.e. smaller somata interspersed between the peripherally projecting DUM-cells are not octopamine immunoreactive, but, confirming other studies 25, display GABA-like immunoreactivity. We thus suggest, that of the whole DUM-cell population in the examined ganglia, all and only peripherally projecting DUM-neurons are octopaminergic.

Octopamine immunoreactive cell populations in the locust thoracic-abdominal nervous system.

We describe octopamine-immunoreactive somata and their projections in the pro- meso-, meta- and pregenital abdominal-ganglia of locusts. Immunoreactive midline somata were identified as dorsal- and ventral- unpaired median (DUM- and VUM-, respectively) neurones due to their: characteristic large size and positions of somata, primary neurites in DUM-tracts giving rise to T-junctions, and bilaterally projecting axons. In the prothoracic ganglion there are most likely 8 such cells; in the meso- and metathoracic, some 20 each; and in each individual pregenital abdominal ganglion, typically 3. All appear to project to peripheral nerves and their numbers correspond to the number of peripherally projecting DUM-cells identified to date in each ganglion. We suggest that probably all peripherally projecting DUM-cells are octopaminergic in the examined ganglia. Presumptive DUM-interneurones are not octopamine-immunoreactive, but, confirming other studies, are shown to label with an antiserum to gamma-amino butyric acid (GABA). Other octopamine-immunoreactive neurones include a pair of midline, prothoracic, anterior medial cells, not necessarily DUM-cells, and a pair of ventral lateral somata in each thoracic- and the first abdominal ganglion. The latter project intersegmentally in ventral tracts. Intersegmentally projecting octopamine-immunoreactive fibers in dorsal tracts probably arise from a prothoracic DUM-cell, which leaves through suboesophageal nerves, or descending suboesophageal DUM-cells. Thus, the octopamine-immunoreactive system of thoracic and pregenital abdominal ganglia in locust comprises all peripherally projecting DUM-cells and a plurisegmental network.

A method for preparation of fibrin glue.

Otorhinolaryngological operations would be facilitated by the use of a safe, cheap, surgical adhesive. Though commercial fibrin glue is effective, it is both relatively expensive and, because it is prepared from pooled human blood, there has been concern that it carries the risk of transmitting viral disease, in particular hepatitis and acquired immune deficiency syndrome. We describe a rapid, cheap method of preparing sufficient fibrin glue, of adequate bonding power, from a small quantity of the patient's own blood, and therefore with no risk of contracting viral disease. Our method of preparation of fibrin glue promises to provide a valuable adjunct to otorhinolaryngological surgical practice.

Bronchiectasis in acute leukaemia.

Five children in remission from acute lymphoblastic leukaemia developed bronchiectasis when on chemotherapy. Persistent collapse or consolidation on chest radiographs was helpful in suggesting the diagnosis. Necropsy established the diagnosis in one child who died of massive haemoptysis when in complete remission, and bronchography confirmed the diagnosis in three. In a further child the diagnosis was based on clinical and chest X-ray findings alone. The surviving children were treated with prophylactic rotating antibiotics. Routine chest radiographs are recommended in children with acute lymphoblastic leukaemia, as bronchiectasis may otherwise be underdiagnosed.