Acute myeloid leukemia (AML) primary cells express high levels of phosphorylated Akt, a master regulator of cellular functions regarded as a promising drug target. By means of reverse phase protein ...arrays, we examined the response of 80 samples of primary cells from AML patients to selective inhibitors of the phosphatidylinositol 3 kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) axis. We confirm that >60% of the samples analyzed are characterized by high pathway phosphorylation. Unexpectedly, however, we show here that targeting Akt and mTOR with the specific inhibitors Akti 1/2 and Torin1, alone or in combination, result in paradoxical Akt phosphorylation and activation of downstream signaling in 70% of the samples. Indeed, we demonstrate that cropping Akt or mTOR activity can stabilize the Akt/mTOR downstream effectors Forkhead box O and insulin receptor substrate-1, which in turn potentiate signaling through upregulation of the expression/phosphorylation of selected growth factor receptor tyrosine kinases (RTKs). Activation of RTKs in turn reactivates PI3K and downstream signaling, thus overruling the action of the drugs. We finally demonstrate that dual inhibition of Akt and RTKs displays strong synergistic cytotoxic effects in AML cells and downmodulates Akt signaling to a much greater extent than either drug alone, and should therefore be explored in AML clinical setting.
ABSTRACT Fibrocartilaginous embolism (FCEM) and acute, non-compressive nucleus pulposus extrusion (ANNPE) are non-compressive myelopathies that are difficult to differentiate. The definitive ...diagnosis is obtained only with histology, but the presumptive diagnosis is made through clinical signs and imaging tests. The aim of this study is to report the imaging tests performed for the diagnosis of a neurological clinical case and discuss the best diagnostic method. After attending the patient, complementary tests were requested. Radiography results showed no change. The computed tomography diagnostic impression indicated distal protrusion between C6-C7, T11-T12, T13-L1 followed by mild spinal cord compression defined by the presence of a ventral hyperattenuating region. Magnetic resonance (RMI), showed a slight T2W hypersignal, well delimited in the gray matter, lateralized to the right, over the cranial third of C7. Concluding that the magnetic resonance is the method that brought more information for the diagnosis, in which the others were not described medullary alterations pertinent to FCEM and ANNPE. With their fair prognosis, the absence of histological diagnosis of these diseases may be a limiting factor in this study and, in relation to the RMI alterations being very similar between FCEM and ANNPE it is not possible to diagnose fully accurately.
RESUMO A embolia fibrocartilaginosa (EFC) e a extrusão aguda não compressiva do núcleo pulposo (EANCNP) são mielopatias não compressivas de difícil diferenciação. O diagnóstico definitivo é obtido apenas com a histologia, mas o diagnóstico presuntivo é feito por meio de sinais clínicos e exames de imagem. O objetivo deste trabalho é relatar os exames de imagem realizados para o diagnóstico de um caso clínico neurológico e discutir o melhor método diagnóstico. Após o atendimento do paciente, foram solicitados exames complementares. Os resultados da radiografia não mostraram nenhuma alteração. A impressão diagnóstica da tomografia computadorizada indicou protrusão distal entre C6-C7, T11-T12, T13-L1, seguida de leve compressão medular definida pela presença de região hiperatenuante ventral. À ressonância magnética (RM), apresentava discreto hipersinal em T2W, bem delimitado na substância cinzenta, lateralizado à direita, sobre o terço cranial de C7. Concluiu-se que a ressonância magnética é o método que mais trouxe informações para o diagnóstico, os demais métodos não foram descritos alterações medulares pertinentes à EFC e à EANCNP. Com seu prognóstico favorável, a ausência de diagnóstico histológico dessas doenças pode ser um fator limitante neste estudo. Em relação às alterações do RM serem muito semelhantes entre EFC e EANCNP, não é possível diagnosticar com total precisão.
Macrophages reside in tissues infiltrated by chronic lymphocytic leukemia B cells and the extent of infiltration is associated with adverse prognostic factors. We studied blood monocyte population by ...flow cytometry and whole-genome microarrays. A mixed lymphocyte reaction was performed to evaluate proliferation of T cells in contact with monocytes from patients and normal donors. Migration and gene modulation in normal monocytes cultured with CLL cells were also evaluated. The absolute number of monocytes increased in chronic lymphocytic leukemia patients compared to the number in normal controls (792 ± 86 cells/μL versus 485 ± 46 cells/μL, P=0.003). Higher numbers of non-classical CD14(+)CD16(++) and Tie-2-expressing monocytes were also detected in patients. Furthermore, we performed a gene expression analysis of monocytes in chronic lymphocytic leukemia patients, showing up-regulation of RAP1GAP and down-regulation of tubulins and CDC42EP3, which would be expected to result in impairment of phagocytosis. We also detected gene alterations such as down-regulation of PTGR2, a reductase able to inactivate prostaglandin E2, indicating immunosuppressive activity. Accordingly, the proliferation of T cells in contact with monocytes from patients was inhibited compared to that of cells in contact with monocytes from normal controls. Finally, normal monocytes in vitro increased migration and up-regulated CD16, RAP1GAP, IL-10, IL-8, MMP9 and down-regulated PTGR2 in response to leukemic cells or conditioned media. In conclusion, altered composition and deregulation of genes involved in phagocytosis and inflammation were found in blood monocytes obtained from chronic lymphocytic leukemia patients, suggesting that leukemia-mediated "education" of immune elements may also include the establishment of a skewed phenotype in the monocyte/macrophage population.
Tau protein is of primary importance for neuronal homeostasis and when hyperphosphorylated (PP-Tau), it tends to aggregate in neurofibrillary tangles, as is the case with tauopathies, a class of ...neurodegenerative disorders. Reversible PP-Tau accumulation occurs in the brain of hibernating rodents and it was recently observed in rats (a non-hibernator) during synthetic torpor (ST), a pharmacological-induced torpor-like condition. To date, the expression of PP-Tau in the rat enteric nervous system (ENS) is still unknown. The present study immunohistochemically investigates the PP-Tau expression in the myenteric plexus of the ileum and colon of normothermic rats (CTRL) and during ST, focusing on the two major subclasses of enteric neurons, i.e., cholinergic and nitrergic.
Results showed that both groups of rats expressed PP-Tau, with a significantly increased percentage of PP-Tau immunoreactive (IR) neurons in ST vs. CTRL. In all rats, the majority of PP-Tau-IR neurons were cholinergic. In ST rats, the percentage of PP-Tau-IR neurons expressing a nitrergic phenotype increased, although with no significant differences between groups. In addition, the ileum of ST rats showed a significant decrease in the percentage of nitrergic neurons. In conclusion, our findings suggest an adaptive response of ENS to very low core body temperatures, with changes involving PP-tau expression in enteric neurons, especially the ileal nitrergic subpopulation. In addition, the high presence of PP-Tau in cholinergic neurons, specifically, is very interesting and deserves further investigation. Altogether, these data strengthen the hypothesis of a common cellular mechanism triggered by ST, natural hibernation and tauopathies occurring in ENS neurons.
Summary Objectives Vocal fold paralysis can have an important impact on a patient's quality of life. The goal of this study was to compare, in terms of vocal improvement and motility recovery, the ...post-vocal treatment results of our patients with unilateral vocal fold paralysis (UVFP) when treatment was started early (within 4 weeks from injury) versus intermediate (from 4 to 8 weeks) or delayed (at least 8 weeks after injury) treatment. Study Design An 11-year retrospective study of patients with UVFP who underwent multidimensional diagnostic-therapeutic assessment. Methods In total, 171 patients with UVFP were included in our study, divided into three groups who underwent early (first group), intermediate (second group), or delayed (third group) voice treatment. All patients underwent voice therapy based on forcible exercises supplemented by manipulations and maneuvers. Results Of the 171 patients with UVFP, 106 (62%) recovered vocal fold motility. Of these 106 patients, 51/78 (65%) were in the first group, 30/49 (61%) in the second group, and 25/44 (56%) in the third group. A significant ( P < 0.0001) reduction in fundamental frequency (Fo) was present in the first group with a manifest improvement in the mean values of Jitter (Jitt%; P = 0.001), Shimmer (Shim%; P < 0.0001), and noise-to-harmonic ratio (NHR; P < 0.0001). A significant ( P < 0.0001) reduction in Fo was found in the second group with a manifest improvement in Jitt% ( P < 0.001), Shim% ( P < 0.0001), and NHR ( P < 0.0001). For the third group, no values were statistically significant apart from the improvement in NHR ( P < 0.001). Conclusions This study confirms the importance of early rehabilitation underlining the non-functional vocal recovery in patients who started treatment later than 8 weeks after injury.
No study has so far addressed whether differences do exist in the management of cancer pain between patients receiving usual care by primary specialists and those receiving early ...palliative/supportive intervention.
A multicentre cross-sectional study in 32 Italian Hospitals has included 1450 patients, receiving analgesic therapy for cancer pain: 602 with access to primary specialist alone (standard care, SC) and 848 with early access to a palliative/supportive care (ePSC) team, concomitant with primary oncology care.
Statistically significant differences in the analgesic drug administration according to care model have been evident: non-opioids were more frequently used in SC (9.5 % versus 2 % ; P < 0.001), while strong opioids in ePSC group (80 % versus 63 % ; P < 0.001). The number of patients with severe pain was lower in ePSC compared with SC group (31 % versus 17 % ; P < 0.001). Results of multivariate analysis have shown that ePSC integrated with primary oncologic care (relative risk 0.69; 95 % confidence interval 0.48–0.99; P = 0.045) was an independent factor associated with a 31 % reduced risk of suffering from severe pain.
An ePSC team provides the most effective standard of analgesic therapy for cancer pain. A randomized clinical trial is needed to confirm these findings.
Primary Effusion Lymphoma (PEL) is an HHV-8-related non Hodgkin lymphoma localized in body cavities (as pleural, peritoneal and pericardial) presenting lymphomatous effusion that, until now, lack of ...an effective therapy. Curcumin was reported to display pro-apoptotic effect via the inhibition of the JAK/STAT pathway, that is overexpressed in PEL cells, as consequence of virus infection. The administration of curcumin is severely restricted by its physicochemical properties, mainly its low solubility in biological fluid and consequently low bioavailability. Encapsulation into biocompatible and biodegradable PLGA nanoparticles (NPs) could be a strategy to overcome biological limits of curcumin, offering a valuable step forward for its clinical application. In this study we described single-emulsion process for curcumin loading into NPs (encapsulation efficiency about 35%). We applied a post-formulation strategy (NHS/EDC reaction) to decorate the surface of the curcumin-loaded NPs with quantum dots (QDs) as imaging agents (QDs-NPs-Cur, 24pmol of QDs per 100mg of NPs) obtaining tools useful for possible application in theranostic approach.
Bifunctionalized NPs were tested in vitro on two PEL's cell line (BCBL-1 and HBL-6). The efficacy of the treatment was evaluated by cytofluorimetric assay by measuring both cell viability and cell density. We found that the NPs significantly improve the cellular effect of curcumin (respect to free drug). Moreover, by means of confocal microscopy, both the localization of bifunctional NPs and of the released drug were easily detectable.
Thus, we conclude that the delivery of curcumin using bifunctional traceable NPs is a promising future approach for the diagnosis and the treatment of PEL.
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After transplant, patient infection with human herpesvirus 8 (HHV‐8) and Kaposi sarcoma–associated herpesvirus (KSHV) is known to cause aggressive tumors and severe nonneoplastic complications. These ...latter syndromes are driven by HHV‐8/KSHV lytic reactivations and related hyperinflammatory host responses typically characterized by high viral loads, elevated levels of cytokines and other inflammation biomarkers, cytopenia, organ failure, high fever, and worsening conditions (with no evidence of B cell neoplasias). These disorders are associated with a high mortality rate, often due to lack of prompt diagnosis, effective therapeutic approaches, and adequate follow‐up. These features resemble most of those defining the so‐called KSHV‐associated inflammatory cytokine syndrome (KICS), which was recently recognized in patients positive for human immunodeficiency virus (HIV). In this report, we describe—for the first time—a case of a KICS‐like nonneoplastic recurrent complication occurring after transplant in an HIV‐negative patient that was successfully treated by a combination of anti‐CD20 monoclonal therapy, antivirals, and modification of the immunosuppressive regimen. In addition to clinical and laboratory findings collected during 3‐year follow‐up, we report novel experimental data on HHV‐8–specific T cell dynamics and circulating microRNA profile, showing correlations with clinical course and other laboratory markers (including viral load, C‐reactive protein, and cytokine levels), providing useful information about abnormal cellular and cytokine dynamics underlying HHV‐8–associated inflammatory disorders in posttransplant patients.
The authors report the first case of successful treatment of Kaposi sarcoma herpesvirus inflammatory cytokine syndrome occurring in a kidney–liver transplant patient secondary to donor‐derived HHV‐8 infection together with experimental data regarding HHV‐8–specific T cell dynamics and circulating miRNA profile.
Tracheostomy is required to ensure a safe airway in open partial horizontal laryngectomies. The presence of the tracheostomy tube can contribute to post-operative dysphagia. This study aimed to ...evaluate the effects of a circumferential tracheostomy technique on swallowing.
A retrospective study was conducted of patients who underwent open partial horizontal laryngectomies between April 2018 and June 2019. Patients were divided into two groups based on the tracheostomy technique: group 1 had two stitches from the inferior tracheal ring to the skin; group 2 had circumferential fixation of the trachea to the skin. Demographic information, surgical data, post-operative rehabilitation course and complication details were collected and analysed.
Twenty-four patients were enrolled. Patients in group 2 had significant improvement in the initial phases of swallowing rehabilitation.
Tracheostomy with anchorage of the trachea to the skin by circumferential stitches could allow early removal of the tracheal tube, with a better swallowing outcome.
Purpose
We investigated the clinical performance of (1 → 3)-β-
d
-glucan (BG), as an early marker of invasive fungal infections (IFI), in different clinical settings.
Methods
BG serum levels were ...assessed by Fungitell (Associates of Cape Cod, Inc), in parallel with galactomannan (GM) when requested by clinicians. By a prospective monocentric study, 270 episodes at risk or with suspect of IFI were enrolled, namely 58 proven-probable invasive aspergillosis (IA), 27 proven invasive candidiasis (IC), 11 possible IC, 16
P.jirovecii
pneumonia (PJP), 4 episodes of other IFI and 154 non-IFI controls.
Results
We found that (a) the BG overall sensitivity, specificity, positive predictive value and negative predictive value (NPV) were 87.9, 80.5, 76.7 and 89.9 %, respectively; (b) the highest sensitivity was found in the IC groups, followed by PJP, IA and other IFI groups; (c) an association was observed between BG kinetics and patients outcome; (d) in the IA episodes, the combination of BG or GM vs GM alone increased sensitivity from 60.0 to 83.3 % in the haematological patients; (e) false-positive BG results were related to Gram-negative infections or infusion of polyclonal IgM-enriched immunoglobulins, where high levels of BG were indeed detected.
Conclusion
Besides strengthening its overall good clinical performance, we provide evidence that serum BG correlates with clinical outcome and that, once used in combination with GM, BG allows to enhance IFI diagnosis rate. The high sensitivity and NPV, observed in the Intensive Care Unit setting, open to BG validation as a marker for assessment of antifungal treatment.
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