Lymphoproliferative disorders (LPDs) associated with inborn errors of immunity (IEI) are rare entities and their genetic basis is not well defined. We performed targeted deep sequencing using a panel of 529 genes to investigate the single nucleotide variants (SNVs), insertion/deletions (INDELs), structural rearrangements (SVs), and copy number variants (CNVs) in 16 lymphoproliferations associated with IEI belonging to a cohort of 14 patients. Histomorphologically, the cases were classified into B-cell hyperplasia (n = 1); polymorphic B-lymphoproliferative disorder (n = 8); polymorphic B-lymphoproliferative disorder with focal increased large cell component bordering large B cell lymphoma (n = 3); and large B-cell lymphoma (n = 4). Fourteen of the 16 cases were EBV positive. Across the patients, 40 variants were identified. The majority of the genes affected were those involved in DNA damage response pathways and transcriptional regulation. Pathogenic SNVs, INDELs, and CNVs were increased in cases with a large B cell component vs other cases. Heterozygous SNVs in protein interaction domains of EMSY were identified in 3 cases, suggesting that it may have a predisposing role in the development of lymphoproliferations. Deletions involving the TNFAIP3 gene (copy number losses) were also recurrent, identified in 3 of 16 cases (18.8

Adenoid cystic carcinoma (ACC) is a common malignant tumor of the salivary glands. However, it lacks established prognostic markers. This study investigates the prognostic relevance of glucose transporter-1 (GLUT-1) in ACC. Seventy patients diagnosed with ACC were included in the study. Immunohistochemical analysis was performed to evaluate GLUT-1 expression levels by intensity, distribution, and scoring. These scores were correlated with clinicopathological parameters, overall survival (OS), and disease-free survival (DFS). The prognostic performance of GLUT-1 was assessed using ROC curve analysis and independent prognostic factors were identified through univariate and multivariate Cox regression models. The GLUT-1 expression rate was 98.5

Background: Multicentric Castleman disease (MCD) is a rare, aggressive lymphoproliferative disorder. Human herpesvirus-8 (HHV-8) has an important role in the pathogenesis of the disease and its association with Kaposi’s sarcoma has been reported, especially in people living with human immunodeficiency virus (HIV). In this report, we present the case of HHV-8 positive MCD accompanied by Kaposi’s sarcoma and multiple myeloma in an HIV-negative patient. Case Report: A 78-year-old man with Kaposi’s sarcoma presented with B symptoms, pancytopenia, lymphadenopathy, and splenomegaly. The bone marrow biopsy demonstrated 70% lambda-restricted monotypic plasma cell infiltration consistent with plasma dyscrasia. Also, the patient was diagnosed with HHV-8 positive MCD as a result of inguinal lymph node excisional biopsy. Treatment was initiated including ganciclovir and methylprednisolone and followed by rituximab. The patient passed away at the 24th hour of rituximab infusion due to shock. Conclusions: MCD and associated malignancies are difficult to treat and have a poor prognosis. More studies and data are needed to manage these patients.

Postmastectomy radiotherapy causes capsular contracture due to fibroproliferation of the capsular tissue around the implant. In fibrosis, unlike normal wound healing, structural and functional disorders are observed in the tissues caused by excessive/irregular accumulation of extracellular matrix proteins. It has been reported that transforming growth factor-beta 3 (TGF-beta 3) prevents and reverses fibrosis in various tissues or provides scarless healing with its antifibrotic effect. Additionally, TGF-beta 3 has been shown to reduce fibrosis in radiotherapy-induced fibrosis syndrome. However, no study in the literature investigates the effects of exogenously applied TGF-beta 3 on capsular contracture in aesthetic or reconstructive breast implant application. TGF-beta 3, which has a very short half-life, has low bioavailability with parenteral administration. Within the scope of this study, free TGF-beta 3 was loaded into the nanoparticles to increase its low bioavailability and extend its duration of action by providing controlled release. The aim of this study is to investigate the preventive/improving effects of radiation induced capsular contracture using chitosan film formulations containing TGF-beta 3 loaded poly(lactic-co-glycolic acid)-b-poly(ethylene glycol) (PLGA-b-PEG) nanoparticles in implant-based breast reconstruction. In the characterization studies of nanoparticles, the particle size and zeta potential of the TGF-beta 3-loaded PLGA-b-PEG nanoparticle formulation selected to be used in the treatment group were found to be 123.60 +/- 2.09 nm and -34.87 +/- 1.42 mV, respectively. The encapsulation efficiency of the formulation was calculated as 99.91 %. A controlled release profile was obtained in in vitro release studies. Chitosan film formulations containing free TGF-beta 3 or TGF-beta 3-loaded PLGA-b-PEG nanoparticles were used in in vivo studies. In animal studies, rats were randomly distributed into 6 groups (n = 8) as sham, implant, implant + radiotherapy, implant + radiotherapy + chitosan film containing unloaded nanoparticles, implant + radiotherapy + chitosan film containing free TGF-beta 3, implant + radiotherapy + chitosan film containing TGF-beta 3 loaded nanoparticle. In all study groups, a 2 cm incision was made along the posterior axillary line at the thoracic vertebral level in rats to reach the lateral edge of the latissimus dorsi. The fascial attachment to the chest wall was then bluntly dissected to create a pocket for the implants. In the treatment groups, the wound was closed after films were placed on the outer surface of the implants. After administering prophylactic antibiotics, rats were subjected to irradiation with 10 Gy photon beams targeted to each implant site. Each implant and the surrounding excised tissue were subjected to the necessary procedures for histological (capsule thickness, cell density), immunohistochemical, and biochemical (alpha-SMA, vimentin, collagen type I and type III, TGF-beta 1 and TGF-beta 3: expression level/protein level) examinations. It was determined that the levels of TGF-beta 1 and TGF-beta 3 collagen type III, which decreased as a result of radiotherapy, were brought to the control level with free TGF-beta 3 film and TGF-beta 3 nanoparticle film formulations. Histological analyses, consistent with biochemical analyses, showed that thick collagen and fibrosis, which increased with radiotherapy, were brought to the control level with free TGF-beta 3 film and TGF-beta 3 nanoparticle film treatments. In biochemical analyses, the decrease in thick collagen was compatible with the decrease in the collagen type I/type III ratio in the free TGF-beta 3 film and TGF-beta 3 nanoparticle film groups. Changes in protein expression show that TGF-beta 3 loaded nanoparticles are more successful than free TGF-beta 3 in wound healing. In line with these results and the literature, it is thought that the balance of TGF-beta 1 and TGF-beta 3 should be maintained to ensure scarless wound healing with no capsule contracture.

Mantle cell lymphoma (MCL) is genetically characterized by the IG::CCND1 translocation mediated by an aberrant V(D)J rearrangement. CCND1 translocations and overexpression have been identified in occasional aggressive B-cell lymphomas with unusual features for MCL. The mechanism generating CCND1 rearrangements in these tumors and their genomic profile are not known. We have reconstructed the IG::CCND1 translocations and the genomic profile of 13 SOX11-negative aggressive B-cell lymphomas using whole genome/exome and target sequencing. The mechanism behind the translocation was an aberrant V(D)J rearrangement in three tumors and by an anomalous IGH class-switch recombination (CSR) or somatic hypermutation (SHM) mechanism in ten. The tumors with a V(D)J-mediated translocation were two blastoid MCL and one high-grade B-cell lymphoma. None of them had a mutational profile suggestive of DLBCL. The ten tumors with CSR/SHM-mediated IGH::CCND1 were mainly large B-cell lymphomas, with mutated genes commonly seen in DLBCL and BCL6 rearrangements in 6. Two cases, which transformed from marginal zone lymphomas, carried mutations in KLF2, TNFAIP3 and KMT2D. These findings expand the spectrum of tumors carrying CCND1 rearrangement that may occur as a secondary event in DLBCL mediated by aberrant CSR/SHM and associated with a mutational profile different from that of MCL.

Background Resection of metastatic hepatic tumors of breast cancer may result in the acceleration of hepatic and extrahepatic tumor progression due to the microenvironmental circulation of chemokines. This study aimed to investigate the effect of hepatectomy on a large panel of chemokines, liver regeneration, and myeloid cell levels in an experimental breast cancer model. Methods The 4T1 breast cancer cells were inoculated, and 30% to 40% hepatectomy was performed. Mice without tumors or only laparotomy (no hepatectomy) served as control groups. After 14 days (short-term) and 21 days (long-term), tissue samples were obtained from the regions near and distant from the resection site. Chemokine levels were evaluated by enzyme-linked immunosorbent assay arrays. Myeloid infiltration in the liver and the primary tumor and hepatic regeneration status were also histopathologically evaluated. Results The levels of pro-tumorigenic chemokines such as CCL2, CCL3, CCL4, and CCL5 were elevated in hepatectomized tumor-bearing animals. This observation was consistent with the presence of hepatic metastases. Liver regeneration and myeloid cell infiltration showed significant differences between the tumor-bearing hepatectomized groups followed in the short and long term. Conclusion Our study showed elevation and variations in chemokines after hepatectomy, with a prominent increase in pro-tumorigenic chemokines. These results can be associated with the acceleration of metastasis after liver resection. However, further prospective studies are required to better define the impact of resection, which may transform the liver into a favorable site for metastasis.

Anti-tumor necrosis factor alpha (TNF- alpha) biological agents can rarely cause sarcoid-like granulomatosis. A 20-year-old woman presented with a 1-month history of painful left upper eyelid swelling. She was on subcutaneous etanercept and methotrexate for 1 year for juvenile idiopathic arthritis. Imaging showed diffuse enlargement of the left and minimal enlargement of the right lacrimal gland. There was no finding in favor of sarcoidosis on systemic evaluation. Incisional biopsy of the left lacrimal gland revealed non-caseating granulomatous dacryoadenitis. The findings showed significant regression 1 month after cessation of Etanercept therapy. To the best of our knowledge, this report illustrates the first case of an isolated granulomatous dacryoadenitis during TNF-alpha antagonist therapy.

Classical BCR/ABL-1-negative myeloproliferative neoplasms (MPN) are hematopoietic stem cell disorders characterized by clonal proliferation of more than one mature myeloid cell lineage. The most common etiology is the constitutive activation of the JAK/STAT signalling pathway, caused by three main driver mutations: JAK2V617F, CALR, and MPL. In this study, we analyzed the distribution of these driver mutations in our series of classical BCR/AB1-negative MPN patients and their correlation with clinical symptoms, bone marrow, and laboratory findings. We also explored the efficacy of using the CAL2 antibody to identify CALR mutations and studied the potential of using EZH2 antibody expression levels as a prognostic indicator. Our study of 78 BCR/ABL1-negative MPN patients found JAK2V617 mutation in 57.7%, CALR mutation in 11.5%, and MPL mutation in 1.3% of cases. Thrombosis was the most common initial symptom, observed in 25% of patients, predominantly in those with JAK2V617F mutation (p= 0.02). CAL2 was positive in nearly all megakaryocytes of CALR mutant cases (7/9). EZH2 H-scores in megakaryocytes were lower in patients with a higher reticulin fibrosis score (p= 0.013), and thrombotic events were more frequently observed in these patients (p= 0.081). Our findings suggest that CAL2 and EZH2 immunohistochemical staining have potential as diagnostic and prognostic surrogate markers for MPN. Nevertheless, presently, the most crucial components in the diagnosis and prognosis of MPN are comprehensive molecular profiling and its alignment with other diagnostic tools.

Follicular lymphoma is a type of systemic lymphomas which constitutes approximately 30-35% of all Non-Hodgkin lymphomas. It typically presents itself in the form of generalized lymphadenopathy, hepatomegaly, splenomegaly and bone marrow involvement. Cutaneous involvement of follicular lymphoma generally appears as skin-coloured to red, violaceous papules or nodules most commonly involving the scalp, trunk and head&neck region. Herein, we would like to present an unusual case of follicular lymphoma which appears as skin-coloured papules prominent upon the both ears and trunk.

The obesity model is predominantly formed by feeding animals that are prone to obesity with high-fat diets, but this dietary intervention is not standardized. It is very difficult to pellet most high-fat diets, as they become rapidly oxidized due to their high-fat content. This study aimed to use calcium soap (Ca-soap) of beef tallow as a source of fat in high-fat mouse diets and to investigate its effects on pellet quality and its effectiveness in an obesity model. In the experiment, three diets were formed: a control diet (CD), a high-fat diet with beef tallow (HFD), and a high-fat diet containing beef tallow and Ca-soap of beef tallow (Ca-HFD). A total of 36 male C57BL/6 J mice aged six weeks were randomly divided into 3 groups: the Control (C), High-Fat (HF), and High-Fat with Ca-soap (Ca-HF) groups. The experiment lasted 20 weeks. Ca-HFD and HFD had higher hardness and pellet durability index (PDI) values than CD. Ca-HFD had a lower peroxide value than that of the HFD. At the end of the study, average body weight, body weight gain, food consumption, and calorie consumption were found to be the highest in the HF group and the lowest in the C group. The highest epididymal white adipose tissue (eWAT) ratio of the mice was seen in the Ca-HF group. Plasma triglyceride and insulin values were higher in the HF group than in the C group. The highest level of steatosis was observed in the HF group. High body weight gain, fat deposition, hypercholesterolemia, hyperglycemia, glucose tolerance, and hepatic steatosis were observed in C57BL/6 J male mice fed the high-fat diets, but only HFD caused hyperinsulinemia. HFD and Ca-HFD can be used to model obesity, but only HFD can be used in type 2 diabetes modeling.

Karaosmanoglu, Ali Devrim MD; Arslan, Sevtap MD; Ozbay, Yakup MD; Sokmensuer, Cenk MD; Ozogul, Ece MD; Karcaaltincaba, Musturay MDAuthor Information

OBJECTIVE:The aim of this study was to investigate the utility of BRCA1-associated protein-1 (BAP1), glucose transporter (GLUT)-1 and desmin expression by immunohistochemistry in the discrimination between reactive and malignant mesothelial proliferations.METHODS:A total of 88 biopsies and 30 effusions from mesothelioma cases were studied. Control groups were composed of 35 tissues and 30 cell blocks. The 88 mesothelioma cases were from 43 males and 45 females (mean age 56 years). Tumours were mostly localised to pleura (66/88, 75%) and of epithelioid histology (75/88, 85%). Cytology samples were from 17 males and 13 females (mean age 58 years), and 16 pleural and 14 peritoneal effusions. Twenty cytology cases had corresponding tissue biopsies.RESULTS:BAP1 loss was detected in 61/88 (69%) tissues and in 20/30 (67%) cytology samples from mesothelioma with a specificity of 100% for both sampling methods. BAP1 loss was observed more frequently in pleural and biphasic tumours. GLUT-1 immunoreactivity was identified in 54/81 (67%) and 23/25 (92%) malignant tissues and effusions, and in 6/33 (18%) and 6/30 (20%) benign tissues and effusions, respectively. Desmin loss was observed in 74/80 (92%) malignant biopsy samples, 16/21 (76%) malignant effusions and 10/34 (29%) of benign tissues, but in none of the reactive effusions. Concordance rate of results between biopsy and cytology was as follows: BAP1 20/20 (100%); GLUT-1 13/18 (72%); and desmin 10/14 (71%).CONCLUSIONS:BAP1, GLUT-1 and desmin are useful markers in the discrimination between reactive and malignant mesothelial proliferations. BAP1 loss seems to be diagnostic for mesotheliomas both in biopsy and cytology samples.

Angioimmunoblastic T-cell lymphoma (AITL) is a common subtype of peripheral T-cell lymphoma, accounting for approximately one-fifth of cases [1]. Epstein-Barr virus (EBV)positive B cells are present in the tumor tissue in most cases [2]. Twenty-five cases of EBV-associated B-cell lymphomas in AITL patients have been reported in the literature [1,3,4,5,6,7,8,9,10, 11,12,13,14,15]; herein, we report the 26th case.

OBJECTIVEProgrammed death ligand 1 (PD-L1) found on tumor cells has recently been reported to have a key role in the development and dissemination of many tumors, such as lung and breast carcinomas. In this study, we retrospectively analyzed PD-L1 expression among different types of sarcomas.MATERIAL AND METHODTissue microarrays of 3-4 mm diameter were composed from paraffin blocks of 222 various sarcomas. Slides prepared from microarrays were stained for PD-L1 antibody (Cell Signaling, E1L3N®) using Leica Bond Autostainer. Any membranous staining over 5% of the cells was regarded as positive. Quantitative real-time PCR with TaqMan gene expression assays for PDL1 was performed using whole sections from FFPE tissue of PD-L1 positive cases, by normalizing absolute values to β-actin. Relative expression level of mRNA of PDL1 was calculated and scored using Log102(threshold cycle of b-actin - threshold cycle of PDL1).RESULTSImmunohistochemically, PD-L1 expression was present in 34 of 222 (15%) sarcomas. 5/13 (39%) undifferentiated pleomorphic sarcomas, 6/18 (33%) malignant peripheral nerve sheath tumors, 5/16 (31%) dedifferentiated liposarcomas, 4/19 (21%) rhabdomyosarcomas, 2/16 (13%) epithelioid sarcomas, 2/15 (13%) leiomyosarcomas, 3/26 (12%) synovial sarcomas, 1/18 (6%) myxoid liposarcoma, 1/2 (50%) extraskeletal myxoid chondrosarcoma, 1/3 (33%) alveolar soft part sarcoma, 1/3 (33%) parachordoma/myoepithelioma, 1/5 (20%) pleomorphic liposarcoma, 1/7 (14%) angiosarcoma, 1/8 (13%) Ewing sarcoma showed PD-L1 expression. Cases of solitary fibrous tumor/hemangiopericytoma (18), desmoplastic round cell tumor (14), Ewing-like sarcoma (6), epithelioid hemangioendothelioma (5), clear cell sarcoma (4), myxofibrosarcoma (4), low grade fibromyxoid sarcoma (2) were all negative. Tumor-infiltrating hematopoietic cells were positive for PD-L1 in 32 cases (15%) with only 2 cases overlapping with PD-L1 staining in tumoral cells. Sixteen of 34 (47%) immunohistochemically PD-L1 positive cases showed significant but low-level PD-L1 mRNA overexpression.CONCLUSIONWe have shown PD-L1 expression in a subset of sarcomas, both at the protein and mRNA level. High-grade pleomorphic sarcomas tend to show more frequent PD-L1 expression. Clinical trials are necessary to further assess the effect of anti PD-L1 drugs on sarcomas showing PD-L1 expression.

BRCA1-associated protein 1 (BAP1) gene is located at chromosome region 3p21.1, a genomic region that is deleted in several human malignancies, including approximately 30-60% of mesotheliomas(1). In this study, we retrospectively investigated BAP1 status in 41 unrelated patients with mesothelioma who had a history of environmental fibrous mineral exposure (erionite or asbestos). We have also reviewed histological type and clinical characteristics of the analyzed patients. A total of 41 malignant mesothelioma cases were reviewed histopathologically. Representative areas were selected and 4-mm-diameter tissue microarrays were composed from paraffin blocks. Immunohistochemistry (IHC) was performed on paraffin tissue sections prepared from microarrays with a monoclonal antibody against BAP1. Cases with loss of nuclear staining were considered as loss of BAP1 expression. Satisfactory results were obtained in 37 patients (25 females, 12 males; mean age 56 yrs). Thirty-one cases were pleural, 5 cases were peritoneal and 1 case was paratesticular mesotheliomas. Histologically, 31 cases were epithelioid, and 6 cases were biphasic type. Overall all loss of BAP1 expression was 31/37 [83, 8% (87, 1% pleural, 80% peritoneal, 0 paratesticular)]. Histologically, all biphasic types and 25/31 (80, 6%) epithelioid types showed BAP1 expression loss. Loss of BAP1 expression seems to be a frequent event in Turkish malignant mesotheliomas. However, in our small cohort, no significant correlation was found between tumor type and localization. We need to demonstrate both somatic and germ-like mutations in familial cases especially from erionite villages. References:1. Carbone M. BAP1 and Cancer. Nat Rev Cancer; 13: 153–159,1.2013.

We report the case of a 7-year-old boy who presented with a swollen right eye. Magnetic resonance imaging revealed a right intraconal orbital mass with intense contrast enhancement. Incisional biopsy led to a diagnosis of perivascular epithelioid cell tumor (PEComa). Sirolimus was initiated but discontinued at the third week of treatment because the tumor had progressed. A minor regression of the tumor was seen after six cycles of systemic chemotherapy. Previously reported cases of PEComa were benign in nature, and full remission was achieved with surgical excision. In the present case the tumor was malignant and responded only slightly to systemic chemotherapy.