Targeting Melanoma-Associated Fibroblasts (MAFs) with Activated γδ (Vδ2) T Cells: An In Vitro Cytotoxicity Model.

The tumor microenvironment (TME) has gained considerable scientific attention by playing a role in immunosuppression and tumorigenesis. Besides tumor cells, TME is composed of various other cell types, including cancer-associated fibroblasts (CAFs or MAFs when referring to melanoma-derived CAFs) and tumor-infiltrating lymphocytes (TILs), a subpopulation of which is labeled as γδ T cells. Since the current anti-cancer therapies using γδ T cells in various cancers have exhibited mixed treatment responses, to better understand the γδ T cell biology in melanoma, our research group aimed to investigate whether activated γδ T cells are capable of killing MAFs. To answer this question, we set up an in vitro platform using freshly isolated Vδ2-type γδ T cells and cultured MAFs that were biobanked from our melanoma patients. This study proved that the addition of zoledronic acid (1-2.5 µM) to the γδ T cells was necessary to drive MAFs into apoptosis. The MAF cytotoxicity of γδ T cells was further enhanced by using the stimulatory clone 20.1 of anti-BTN3A1 antibody but was reduced when anti-TCR γδ or anti-BTN2A1 antibodies were used. Since the administration of zoledronic acid is safe and tolerable in humans, our results provide further data for future clinical studies on the treatment of melanoma.

Mesenchymal-Stromal Cell-like Melanoma-Associated Fibroblasts Increase IL-10 Production by Macrophages in a Cyclooxygenase/Indoleamine 2,3-Dioxygenase-Dependent Manner.

Melanoma-associated fibroblasts (MAFs) are integral parts of melanoma, providing a protective network for melanoma cells. The phenotypical and functional similarities between MAFs and mesenchymal stromal cells (MSCs) prompted us to investigate if, similarly to MSCs, MAFs are capable of modulating macrophage functions. Using immunohistochemistry, we showed that MAFs and macrophages are in intimate contact within the tumor stroma. We then demonstrated that MAFs indeed are potent inducers of IL-10 production in various macrophage types in vitro, and this process is greatly augmented by the presence of treatment-naïve and chemotherapy-treated melanoma cells. MAFs derived from thick melanomas appear to be more immunosuppressive than those cultured from thin melanomas. The IL-10 increasing effect is mediated, at least in part, by cyclooxygenase and indoleamine 2,3-dioxygenase. Our data indicate that MAF-induced IL-10 production in macrophages may contribute to melanoma aggressiveness, and targeting the cyclooxygenase and indoleamine 2,3-dioxygenase pathways may abolish MAF-macrophage interactions.

The Human Melanoma Proteome Atlas-Complementing the melanoma transcriptome.

The MM500 meta-study aims to establish a knowledge basis of the tumor proteome to serve as a complement to genome and transcriptome studies. Somatic mutations and their effect on the transcriptome have been extensively characterized in melanoma. However, the effects of these genetic changes on the proteomic landscape and the impact on cellular processes in melanoma remain poorly understood. In this study, the quantitative mass-spectrometry-based proteomic analysis is interfaced with pathological tumor characterization, and associated with clinical data. The melanoma proteome landscape, obtained by the analysis of 505 well-annotated melanoma tumor samples, is defined based on almost 16 000 proteins, including mutated proteoforms of driver genes. More than 50 million MS/MS spectra were analyzed, resulting in approximately 13,6 million peptide spectrum matches (PSMs). Altogether 13 176 protein-coding genes, represented by 366 172 peptides, in addition to 52 000 phosphorylation sites, and 4 400 acetylation sites were successfully annotated. This data covers 65% and 74% of the predicted and identified human proteome, respectively. A high degree of correlation (Pearson, up to 0.54) with the melanoma transcriptome of the TCGA repository, with an overlap of 12 751 gene products, was found. Mapping of the expressed proteins with quantitation, spatiotemporal localization, mutations, splice isoforms, and PTM variants was proven not to be predicted by genome sequencing alone. The melanoma tumor molecular map was complemented by analysis of blood protein expression, including data on proteins regulated after immunotherapy. By adding these key proteomic pillars, the MM500 study expands the knowledge on melanoma disease.

The human melanoma proteome atlas-Defining the molecular pathology.

The MM500 study is an initiative to map the protein levels in malignant melanoma tumor samples, focused on in-depth histopathology coupled to proteome characterization. The protein levels and localization were determined for a broad spectrum of diverse, surgically isolated melanoma tumors originating from multiple body locations. More than 15,500 proteoforms were identified by mass spectrometry, from which chromosomal and subcellular localization was annotated within both primary and metastatic melanoma. The data generated by global proteomic experiments covered 72% of the proteins identified in the recently reported high stringency blueprint of the human proteome. This study contributes to the NIH Cancer Moonshot initiative combining detailed histopathological presentation with the molecular characterization for 505 melanoma tumor samples, localized in 26 organs from 232 patients.

Melanoma-associated fibroblasts impair CD8+ T cell function and modify expression of immune checkpoint regulators via increased arginase activity.

This study shows that melanoma-associated fibroblasts (MAFs) suppress cytotoxic T lymphocyte (CTL) activity and reveals a pivotal role played by arginase in this phenomenon. MAFs and normal dermal fibroblasts (DFs) were isolated from surgically resected melanomas and identified as Melan-A-/gp100-/FAP+ cells. CTLs of healthy blood donors were activated in the presence of MAF- and DF-conditioned media (CM). Markers of successful CTL activation, cytotoxic degranulation, killing activity and immune checkpoint regulation were evaluated by flow cytometry, ELISPOT, and redirected killing assays. Soluble mediators responsible for MAF-mediated effects were identified by ELISA, flow cytometry, inhibitor assays, and knock-in experiments. In the presence of MAF-CM, activated/non-naïve CTLs displayed dysregulated ERK1/2 and NF-κB signaling, impeded CD69 and granzyme B production, impaired killing activity, and upregulated expression of the negative immune checkpoint receptors TIGIT and BTLA. Compared to DFs, MAFs displayed increased amounts of VISTA and HVEM, a known ligand of BTLA on T cells, increased L-arginase activity and CXCL12 release. Transgenic arginase over-expression further increased, while selective arginase inhibition neutralized MAF-induced TIGIT and BTLA expression on CTLs. Our data indicate that MAF interfere with intracellular CTL signaling via soluble mediators leading to CTL anergy and modify immune checkpoint receptor availability via L-arginine depletion.

Tardive Dyskinesia Associated with Bupropion.

Present report describes a 46 year old male patient with a diagnosis of major depression who developed tardive dyskinesia during bupropion therapy. Our patient had no history of neuroleptic use and his laboratory and neurologic examinations were normal. He had no family history of neurologic diseases. Although bupropion induced dyskinesia has been previously reported in the literature, it is rare and our case is the first case regarding tardive dyskinesia.

A new diagnostic tool for masked hypertension: impaired sleep quality.

INTRODUCTION: The aim of this study was to evaluate the relationship between masked hypertension and impaired sleep quality. Additionally, we evaluated the diagnostic role and prevalence of poor sleep quality among patients with newly diagnosed masked hypertension. MATERIAL AND METHODS: A total of 112 individuals, 72 patients with newly diagnosed masked hypertension and 40 normotensive healthy volunteers, were included in this study. All patients underwent evaluation comprising 12-lead electrocardiography, transthoracic echocardiography, 24-hour Holter ECG, and basic laboratory tests. Additionally, all participants completed questionnaires, including the Pittsburgh Sleep Quality Index (PSQI). RESULTS: The total PSQI score was significantly higher in the masked hypertension group than in the normotensive healthy volunteers (4.13 ±2.43 vs. 2.33 ±1.67, p < 0.001). A PSQI score > 5 was found in 45.8% (n = 33) of patients in the masked hypertension group and 15% (n = 6) of patients in the normotensive group (p < 0.001). The non-dipper pattern was found in 17.5% of the healthy volunteer group and 59.94% (n = 41) of the masked hypertension group (p < 0.001). When we compared the dipping pattern of the masked hypertension groups, there was a significant difference in PSQI score between the dipper and non-dipper groups (4.87 ±3.21 vs. 3.58 ±2.33, p < 0.001). Multiple logistic regression analyses showed that masked hypertension, LV mass, and LV mass index score were independent predictors of poor PSQI. CONCLUSIONS: This study demonstrates impaired sleep quality in subjects with masked hypertension, particularly those with a non-dipper pattern. Additionally, this study indicates that impaired sleep quality may help diagnose masked hypertension, particularly in the non-dipper group.

Leukocytosis after Clozapine Treatment in a Patient with Chronic Schizophrenia.

Clozapine is an atypical antipsychotic drug that is approved by the US Food and Drug Administration (FDA) for the treatment of psychotic disorders. Agranulocytosis is a well-established side effect of clozapine; clozapine has also been associated with other blood dyscrasias like leukocytosis, albeit rarely. In this paper, we aim to report a case of possible clozapine-associated leukocytosis in a 41-year-old woman.

Burden on Caregivers of Patients with Schizophrenia and Related Factors.

INTRODUCTION: Caregivers of patients with schizophrenia are under the burden of continuous and difficult processes. Determination of the factors related to caregiver burden in schizophrenia may help find strategies to decrease the burden. This study aimed at investigating the factors associated with caregiver burden among relatives of patients with schizophrenia. METHODS: Eighty-eight caregivers of patients under treatment for schizophrenia for at least 1 year were included in the study. The Zarit Caregiver Burden Interview was used for the assessment of caregiver burden. Sociodemographical data, the level of knowledge about schizophrenia, clinical impression scale, and global assessment of functioning were used to evaluate the related factors. RESULTS: Caregiver burden was negatively correlated with income level and functionality of the patient and was positively correlated with the age of the caregiver, the daily time spent with the patient, and the number of hospitalizations of the patient (p<0.05). There was no significant correlation between the caregivers' knowledge about schizophrenia and caregiver burden (p<0.05). Living in the same house with the patient was a positive predictor, whereas functionality and income level of the patient and education level of the caregiver were negative predictors (p<0.05). CONCLUSION: This study highlighted the importance of setting targets for improving the functionality of patients in the design and implementation of rehabilitation and support programs for patients with schizophrenia. Additionally, providing higher income for patients, creating conditions for an independent life, and increasing incentives for younger caregivers with a higher educational level may help decrease caregiver burden.

Effects of minimally invasive decompression surgery on quality of life in older patients with spinal stenosis.

OBJECTIVES: Lumbar spinal stenosis (LSS) in the elderly may result in a progressive narrowing of the spinal canal leading to compression of nerve roots in some individuals. The aim of this study was to evaluate the quality of life changes after minimally invasive decompression surgery without instrumentation in geriatric patients with lumbar spinal stenosis. PATIENTS AND METHODS: This prospective clinical study included 37 patients with American Society of Anesthesiologists (ASA) II-III scores between the ages of 65 and 86 years, who were planned to undergo surgical intervention due to LSS. All patients had neurogenic claudication and pain in the hips, thighs, and legs. Measurements of the osseous spinal canal were evaluated by magnetic resonance imaging. Before the surgical intervention, patient demographics and clinical characteristics were recorded. The Short-Form-36 test, the Oswestry Disability Index, and the Visual Analog Scale were applied to all patients preoperatively and two years postoperatively. RESULTS: In the study population, 11 patients had single level of spinal stenosis, 20 patients had two levels of spinal stenosis, and six patients had three levels of spinal stenosis. There were significant differences between the preoperative and postoperative ODI and VAS scores. There was a statistically significant difference in all subscales of the SF-36 test with the exception of general health scores. Three patients who had dural damage during the operation were treated with bio glue. Also, no patients were recorded to have any neurological deficits and root injuries postoperatively. CONCLUSION: Minimally invasive decompression surgery, without instrumentation, for lumbar spinal stenosis in geriatric patients significantly improves the patients' quality of life.

Increased neutrophil/lymphoctye ratio in patients with bipolar disorder: a preliminary study.

BACKGROUND: In this study, it has been aimed to investigate whether neutrophil-lymphocyte ratio (NLR) was higher in non-obese patients with bipolar disorder (BD) than in a healthy control group matched for age, sex, and body mass index, and also to determine if there was an interaction between NLR and severity of the bipolar disorder. SUBJECTS AND METHODS: In this retrospective study, 103 non-obese patients with BD and 126 healthy control subjects were analyzed for complete blood count. The Young Mania Rating Scale (YMRS) was used to determine the severity of the disorder. RESULTS: The NLR was higher in female patients than in female comparison subjects (3.2±2.2; versus 1.7±0.4) (p<0.001). Also, compared with the healthy male subjects, the male patients had significantly higher neutrophil/lymphocyte ratio (3.3±2.4; versus 2.0±0.7) (p<0.001). In the patients with bipolar disorder, NLR did not significantly correlate with severity (as measured with the YMRS) (r=0.052; p=0.204) and duration of the disorder (r=0.045; p=0.301). CONCLUSIONS: Results of this study revealed that patients with bipolar disorder have statistically significant elevated NRL than healthy compares. According to this finding, elevated levels of NLR may be involved in inflammatory pathophysiology of bipolar disorder. Further studies are needed for a better understanding of the mechanism between elevation of NRL in patients with bipolar disorder.

The prevalence of post-traumatic stress disorder in patients with burn injuries, and their quality of life.

OBJECTIVE: The aim of this study was to investigate the prevalence of post-traumatic stress disorder (PTSD) in patients with burn injuries undergoing physical therapy, and to evaluate their quality of life. METHODS: A total of 21 patients who underwent physical therapy for burn injuries between October 2012 and December 2012, in the Physical Therapy and Rehabilitation outpatient clinic of a Training and Research Hospital, were included in the study. The sociodemographic form for data collection, the Clinician- Administered PTSD Scale (CAPS) for the diagnosis of PTSD, and the Short Form 36 (SF-36) Health Survey for the assessment of the quality of life, were used. RESULTS: Eight patients (38.1%) had PTSD. These patients had poor physical functioning, and indicated a lower rate of role functioning-physical, vitality, and role functioning-social, compared to those without PTSD. However, it did not reach statistical significance. The physical functioning related to the quality of life was statistically significantly lower in the patients with contracture. CONCLUSIONS: PTSD seems to be an important health issue in patients with burn injuries. Clinicians who attempt to tailor treatment interventions should keep in mind that these patients require psychosocial rehabilitation, as well as physical therapy.