Bone marrow-resident memory T cells survive pretransplant chemotherapy and contribute to early immune reconstitution of patients with acute myeloid leukemia given mafosfamide-purged autologous bone marrow transplantation

OBJECTIVE: Studies of memory T cells transferred with the graft are relevant to better understand the early immune reconstitution of patients given autologous bone marrow transplantation (A-BMT). A critical question is whether memory T cells resident in bone marrow (BM) of patients with hematological malignancies are resistant to either pretransplant chemotherapy or ex vivo pharmacological purging. PATIENTS AND METHODS: To address these issues, we evaluated the frequency of tetanus-toxoid (TT)-specific proliferating T-cell precursors (TT-PTCp) in BM and peripheral blood (PB) of eight patients with acute myeloid leukemia (AML) given A-BMT after in vitro purging of BM with mafosfamide. Patients were studied at the time of BM harvesting and five of them also after A-BMT. RESULTS: The range of TT-PTCp frequencies found after A-BMT were comparable with those observed in PB and in BM at the time of harvesting and did not differ significantly from those of eight age-matched healthy subjects who donated BM for a human leukocyte antigen-identical sibling. TT-PTCp frequencies in BM, studied before and after ex vivo purging, appeared not to be affected by incubation with mafosfamide. We also compared the T-cell receptor (TCR)-Vbeta-repertoire usage of TT-specific T-cell lines (TT-TCL) in BM of patients at the time of harvesting and in their PB 2 months after transplantation. The same TCR-clonotypes were detected in TT-TCL at time of harvesting and after A-BMT. CONCLUSION: These data indicate that BM-resident memory T cells of patients with AML are resistant to both pretransplant chemotherapy and ex vivo pharmacological purging and may contribute to immune reconstitution after A-BMT.     

Regulation of glutamate release by heteromeric nicotinic receptors in layer V of the secondary motor region (Fr2) in the dorsomedial shoulder of prefrontal cortex in mouse

We studied how nicotinic acetylcholine receptors (nAChRs) regulate glutamate release in the secondary motor area (Fr2) of the dorsomedial murine prefrontal cortex, in the presence of steady agonist levels. Fr2 mediates response to behavioral situations that require immediate attention and is a candidate for generating seizures in the frontal epilepsies caused by mutant nAChRs. Morphological analysis showed a peculiar chemoarchitecture and laminar distribution of pyramidal cells and interneurons. Tonic application of 5 µM nicotine on Layer V pyramidal neurons strongly increased the frequency of spontaneous glutamatergic excitatory postsynaptic currents. The effect was inhibited by 1 µM dihydro‐β‐erythroidine (which blocks α4‐containing nAChRs) but not by 10 nM methyllicaconitine (which blocks α7‐containing receptors). Excitatory postsynaptic currents s were also stimulated by 5‐iodo‐3‐[2(S)‐azetidinylmethoxy]pyridine, selective for β2‐containing receptors, in a dihydro‐β‐erythroidine ‐sensitive way. We next studied the association of α4 with different populations of glutamatergic terminals, by using as markers the vesicular glutamate transporter type (VGLUT) 1 for corticocortical synapses and VGLUT2 for thalamocortical projecting fibers. Immunoblots showed higher expression of α4 in Fr2, as compared with the somatosensory cortex. Immunofluorescence showed intense VGLUT1 staining throughout the cortical layers, whereas VGLUT2 immunoreactivity displayed a more distinct laminar distribution. In Layer V, colocalization of α4 nAChR subunit with both VGLUT1 and VGLUT2 was considerably stronger in Fr2 than in somatosensory cortex. Thus, in Fr2, α4β2 nAChRs are expressed in both intrinsic and extrinsic glutamatergic terminals and give a major contribution to control glutamate release in Layer V, in the presence of tonic agonist levels. Synapse 00:000–000, 2013 . © 2013 Wiley Periodicals, Inc.     

Glassy cell carcinoma of the endometrium: a case report and review of the literature

Glassy cell carcinomas are composed of malignant cells showing a "ground glass" cytoplasm, distinct cell membranes, and large nuclei with prominent nucleoli. To our knowledge, only 12 cases of glassy cell endometrial carcinomas (EGCC) have been reported until now. A 63-year-old patient complaining of irregular vaginal bleeding underwent hysteroscopy-guided biopsy revealing a well-differentiated endometrial endometrioid adenocarcinoma. The patient underwent left salpingo-oophorectomy, total abdominal hysterectomy, and pelvic lymphadenectomy. The final diagnosis was FIGO stage IB poorly differentiated endometrial adenosquamous carcinoma with > 90% of glassy tumor cells. The patient is alive, with no evidence of disease for 69 months after diagnosis. We describe an additional case of EGCC and review the data of the literature, emphasizing the need to strictly define the criteria for the diagnosis and the potential usefulness of assessing biologic parameters for the prognostic characterization of this rare entity.     

Role of relaxation and specific suggestions in hypnotic emotional numbing.

The cognitive characteristics of highly hypnotizable subjects (Highs) allow them to easily modify their cognitive and autonomic state. Under hypnosis, Highs receiving cognitive, fear-like stimulation exhibit the cardiovascular changes typical of fear/stress, but also show an EEG pattern indicating a balance between fear-induced arousal and hypnotic relaxation. Indeed, hypnosis is effective in the attenuation of both emotional experience and behaviour (emotional numbing). The aim of the present experiment was to investigate the possible different role of relaxation and suggestion in hypnotic emotional numbing. Tonic skin conductance, respirogram, heart rate, systolic and diastolic blood pressure were recorded in 3 groups of hypnotized subjects: Group 1 received a fearful guided imagery associated with threat suggestions (Threat) followed by the same fearful suggestion associated with numbing instructions (relaxation and "No-Threat"); Group 2 received the same instructions in the opposite order of presentation; Group 3 received the fearful suggestion with threat instructions twice. The numbing suggestion reduced fear-related emotional experience and autonomic responses; if No-Threat preceded Threat, the heart rate, heart rate variability and blood pressure were also reduced during Threat, in spite of self reports of high negative emotion. Thus, 1) the subjective experience and the autonomic response to fear can be dissociated; 2) the efficacy of numbing suggestion is extended to a subsequent Threat stimulation; 3) habituation does not contribute to the numbing effect. The results indicate that the specific numbing suggestion is the main factor in hypnotic modulation of the experience of fear.     

Imagery of different sensory modalities: hypnotizability and body sway

Postural control in subjects with high (Highs) and low (Lows) susceptibility to hypnosis is differentially affected by changes in visual and neck tactile/proprioceptive input. The aim of the present experiment was to investigate whether imagery of the visual and tactile sensory modalities also induces different modulation of postural control in Highs and Lows. Fourteen Highs and 16 Lows were included in the study; they were recorded while standing upright with eyes closed during visual and tactile imagery tasks and during mental computation. Their posture and movement were recorded with an Elite System and their experience was assessed after each task in a structured interview. Visual imagery was judged “easier” than tactile imagery by Lows, while Highs performed both tasks easily and judged the tactile imagery less effortful and more vivid than Lows. No difference was observed for the mental computation. The Highs’ body sway was not affected by the cognitive tasks, while Lows showed a task-related modulation of body sway. The results are in line with the hypothesis of lower vulnerability of Highs to the effects of tasks interfering with postural control and of different sensory-motor integration in Highs and Lows.     

Fatigue in the masseter and temporalis muscles at constant load

Fatigue is usually defined as the point at which a particular level of force can be no longer maintained. In the present study, surface EMG of the masseter and temporalis anterior muscles was measured in ten healthy young adults performing a unilateral molar (right side) clench. The subjects clenched on a bite force transducer at a fixed force level of 13 kg (127 N) as long as they could (endurance). The test ended when the subjects could no longer produce the required bite force. From the EMG recordings, the median power frequency was calculated at the beginning of the task (T0), after one minute of clenching (T1), and at the end of the task (T2, endurance time). For each subject and muscle, percentage decrements in the median power frequency were also computed at T1 and T2. Endurance time ranged between 79 and 470 s. Significant modifications in the median power frequency in both masseter muscles (right side, p=0.003; left side, p=0.02, analysis of variance) were found, with a significant difference for the median frequency at T2 (p<0.02 at post hoc test). The modifications in the temporalis muscles were not significant (p>0.05). Additionally, at T1, significant percentage decrements in the median power frequency were found for both right side muscles (p<0.05, paired Student's ). The left side muscles modifications (p>0.05) were not significant. A significant effect of side was found (p=0.007, analysis of variance), without effects of muscle and no muscle x side interaction. At T2, both masseter muscles and the right side temporalis had a significant modification in their median power frequency. Overall, the modifications were larger in the masseter than in the temporalis muscles (p=0.022, analysis of variance), without effects of side and no muscle x side interaction. In conclusion, a fixed submaximal muscular contraction provoked fatigue modifications in the EMG power spectra that were well comparable to those obtained in previous investigations using forces computed as percentages of individually assessed maximum bite forces The present protocol (endurance clenching at a fixed force level) could be used as both a research and a clinical tool that would allow an easier and less troublesome assessment of both healthy persons and patients.