Nodular Aggregates Research Articles

The Prognostic Significance of Tumoral Melanosis.

Tumoral melanosis (TM) is a histological term to describe a nodular aggregation of macrophages containing melanin pigment (melanophages) that is devoid of viable melanocytes. It is most often identified in skin, where it may be appreciated clinically as a pigmented lesion; however, it can also be found in other organs such as lymph nodes. The presence of TM is usually thought to signify the presence of a regressed melanoma or other pigmented tumor. Until recently, it was a relatively uncommon finding; however, with the use of effective systemic therapies against melanoma, its occurrence in histological specimens is more frequent. We identified and reviewed all histopathological diagnoses of TM at any organ site reported at a single institution from 2006 to 2018. TM cases were paired with non-TM cases of cutaneous melanoma through propensity score matching at a 1:2 ratio, and their survival outcomes were compared. The clinical outcomes examined included recurrence-free survival (RFS), distant disease-free survival (DDFS), melanoma-specific survival (MSS), and overall survival (OS). TM was reported in 79 patients. Their median age was 65 years (range 22-88), with a 2:1 male predominance (51 out of 79, 65%). The most common organ involved was the skin (67%), with a third of all cases localized to a lower limb (36%). TM had a strong association with the presence of melanoma (91%) and regression at other sites of melanoma (54%), suggesting that it is part of a systemic immune response against melanoma. Most patients with TM either previously or subsequently developed histologically confirmed melanoma in the same anatomical region as the TM (89%). Thirty-five TM patients were matched with 70 non-TM cases. Patients with melanoma who developed TM without prior regional or systemic therapy showed improved MSS (p = 0.03), whereas no statistically significant differences were observed in terms of RFS, DDFS, and OS. TM usually occurs in the context of a previous or subsequent cutaneous melanoma and is associated with improved MSS. It is important that TM is recognized by pathologists and documented in pathology reports.

BRCA1-associated-protein-1 inactivated melanocytic tumours: characterisation of the clinicopathological spectrum and immunohistochemical expression pattern of preferentially expressed antigen in melanoma.

BRCA1-associaed protein-1 (BAP1) inactivated tumours (BIMT) are rare melanocytic tumours that may be mistaken for Spitz tumours or melanoma. They occur sporadically or in association with the BAP1 tumour predisposition syndrome (BAP1-TPDS), which may be complicated by uveal or cutaneous melanoma, mesothelioma, basal cell carcinoma and renal cell carcinoma. The aim of this study was to characterise the clinicopathological features and the immunohistochemical expression pattern of preferentially expressed antigen in melanoma (PRAME) of BIMT in a large patient cohort. Ethical approval was obtained, haematoxylin and eosin-stained slides were reviewed, PRAME immunohistochemistry was performed and clinical follow-up was obtained from patient records. Sixty-five BIMT from 38 patients (F:M = 4.4:1) were identified. BIMT were typically located on the trunk and head and neck (median size = 0.5 cm). Seven patients with BAP1-TPDS (range = 16-66 years, median = 25) had multiple BIMT (median = 5), while sporadic BIMT were solitary (median patient age = 39 years). One of seven patients with BAP1-TPDS developed additional malignancies (mesothelioma and cutaneous spindle cell melanoma) and died of complications of mesothelioma. All other patients are alive without recurrence of BIMT (median follow-up = 42 months). BIMT presented as intradermal, nodular aggregates of epithelioid melanocytes with low mitotic activity and moderate to severe cytological atypia in 63% of cases. A background conventional naevus was present in 64%. PRAME immunohistochemistry showed negative or weakly patchy positive staining in all BIMT. BIMT are more common in a sporadic setting and behave indolently, despite worrying cytological atypia. PRAME immunohistochemistry is a reassuring tool in distinguishing BIMT from melanoma.

A Rare Placental Site Nodule Involving the Ovary.

Placental site nodule (PSN) is a benign lesion representing a nodular aggregate of intermediate trophoblast, embedded in a hyalinized stroma, thought to arise from noninvoluted placental site remaining from a past gestation. Uterus is the most common site of PSN. Occurrence in extrauterine sites is rare, with most examples being reported in the fallopian tubes. Here we report an example of PSN in the ovary. A 35-year-old woman, gravida 4, para 1, with history of adnexal ectopic pregnancy treated with methotrexate, at 39 weeks and 1 day of a subsequent pregnancy, underwent a scheduled C-section. The surgery was successful, and a healthy female infant was delivered. Intraoperative adnexal inspection revealed a small pedunculated mass on the right ovary, which was excised and sent for pathological examination. Gross inspection showed a soft, tan-white tissue fragment measuring 2.0 × 1.0 × 0.2 cm. Microscopic examination showed epithelioid cells with hyperchromatic, mildly atypical nuclei and abundant eosinophilic cytoplasm, embedded in a hyalinized stroma, forming a nodule. A diagnosis of placental site nodule was made. Immunohistochemical stains for keratin AE1/AE3, vimentin, and inhibin were strongly positive in the epithelioid cells, and immunostain for p63 was focally positive, supporting the diagnosis. PSN of the ovary is extremely rare. To our knowledge, there has been only one reported patient in the literature so far. Extrauterine PSNs are thought to arise from ectopic pregnancies. Our patient's ovarian PSN is most likely a consequence of her previous adnexal ectopic pregnancy, which was treated medically.

Mineralogical Characteristics and Genetic Types of Pyrite with Different Occurrence: Constraints from Spectroscopy, Geochemistry and δ34S Stable Isotopes

Pyrite is widely distributed in the Earth’s crust and oceanic systems, and it generally occurs as pentagonal dodecahedra, cubic, octahedral, idiomorphic crystals, or dense, massive, granular, and nodular aggregates. In this study, representative pyrite samples from Hunan, Fujian, Jiangxi, Anhui in China and from Peru were collected. By utilizing a range of analytical techniques, including petrography, X-ray diffraction, infrared spectrum, Raman spectrum, major and trace element analysis, as well as sulfur stable isotope analysis, we comprehensively depict the mineralogical and spectroscopic characteristics of pyrite, and the evolution processes of the physical and chemical conditions of mineralization can be qualitatively constrained. The spectroscopic results indicate all samples show a relatively narrow absorption band with weak to moderate intensity in the vicinity of 343 cm−1, which represents the bending vibration of the Fe-[S2]2− molecular bond. The Co content of pyrite exhibits the characteristics of a positive correlation with temperature and a negative correlation with oxygen fugacity, respectively. The δ34S isotopic compositions of colloidal pyrite are in the range of 0.03 to 0.67, which are close to meteoric sulfur and mantle sulfur compositions, while the δ34S values of nodular pyrite fall within the range of granite, indicating the characteristics of mixtures of sulfur sources are mainly related to magmatic activity. Our results provide insight into the formation mechanisms of pyrite in different environments, its mineralization, and the ore genesis of deposits. Moreover, the integrated analytical methods for pyrite are provided, which can define theoretical guidance for the exploration and development of mineral resources.

Clinical and Histopathologic Study of Apocrine-Type Mixed Tumor of the Skin.

Mixed tumor of the skin (MTS) is a tumor characterized by folliculosebaceous-apocrine differentiation. Because of the wide range of histological variations, understanding the unique features of MTS can help improve diagnosis. This study describes the histopathological characteristics of MTS, mainly apocrine-type MTS (AMT), using 166 cases of AMT. We found that nodular aggregates of myoepithelial cells, mucinous changes in the stroma, and follicular differentiation were standard characteristic features of MTS. Among the cases studied, 67% showed prominent follicular germinative cells and 40% showed prominent lipomatous metaplasia in the stroma. These cases often pose difficulties for the diagnosis of AMT because of insufficient evidence of sweat glands or myoepithelial cell differentiation. This is the first study to examine how the histological features of AMT change as the tumor extends deeper into the dermis. We found that the proportion of AMT with folliculosebaceous differentiation and large lumina increased as it got deeper into the dermis. Histopathological diagnosis of MTS is vital because the clinical symptoms lack specificity. This study enhances our understanding of the histopathological characteristics of MTS.

Study of technological properties of the sample ethiopian iron ore of the arabian-nubian shield. Part 2. Microscopic analysis of ore samples with a thickness of +40.0+5.0 mm

As a result of the research, the mineral composition of the mineralogical and petrological varieties of material of the 10-20 mm and 20-40 mm size classes of the studied ore sample was specified. Transparent and polished sections of the above-mentioned granulometric samples were prepared. Microscopic examination was carried out using a transmitted and reflected light microscope «Carl Zeiss Jena NU-2 Microscope». A description of the discovered minerals was completed. It was found that the main ore minerals of the studied ore are dispersed hematite and hydrohematite, and their amount is about 54.0% of the total volume. Microscopic analysis also revealed hydrohematite spherulites, including in the intergranular space in quartz aggregates. It is shown that secondary ore minerals in the studied raw materials are goethite and hydrogoethite. Microscopic analysis showed their content at the level of 4.0 %. The color of goethite and hydrogethite, which are represented by iron hydroxides of yellow, brown, brown and black color and their form in the form of nodular aggregates, spherulites of crusts, and nest-like aggregates, is also determined. It was found that the main non-metallic mineral in the studied raw material is quartz, the content of which is on average 30 %. This mineral in the sample of the studied ore is characterized by a variety of morphological features and a wide range of grain sizes – from several micrometers to several centimeters. Microscopic examination of quartz showed that it is represented by xenomorphic grains with a complex nature of fusions with other minerals, rounded and acute-angled grains, chalcedony (agat) grains and thin-prismatic crystals. The presence of a secondary non-ore mineral, which has clay properties and is represented by kaolinite, is also shown. An additional study was carried out, in which chalcopyrite – an additional component of iron ore raw materials – was analyzed. The conducted microscopic studies made it possible to draw a conclusion about the mineral and petrological characteristics of the raw material. It is shown that the type of iron ore of the Arabian-Nubian shield is similar to ores of volcanic-sedimentary origin and brown iron ores

Mycosis Fungoides: A Diagnostic Challenge

Background: Mycosis fungoides (MF) is the most frequent type of cutaneous T-cell lymphomas (CTCL), a heterogeneous group of non-Hodgkin lymphoma of T-cell origin. CTCLs accounts for about 4% of all non-Hodgkin lymphoma. MF mainly involves the skin but in advanced cases it may involve the lymph, blood and other organs. It mimics other skin disorders like erythema necroticans, leprosy, psoriasis etc which leads to delayed diagnosis and subsequent treatment. Case report: A 61 year old male patient visited the OPD of a tertiary care hospital in Eastern India with multiple reddish elevated lesions for last 6 months which later became dome shaped and then ulcerated. He did not have any fever or itching. He informed that he had been taking Homeopathic medicine previously, but the identity of the said medicine could not be confirmed. Physical examination ruled out the presence of pallor, jaundice, oedema and cyanosis. Pulse and BP were within normal range. Systemic examination was unremarkable except mild hepatomegaly. Cutaneous examination revealed multiple erythematous indurated plaques, papules and nodules over chest, abdomen, back, limbs and face. Few of the plaques present on the trunk and left arm had developed ulcers having a necrotic floors. Cervical, axillary and inguinal lymph nodes were palpable, being firm in consistency and mobile in nature. Hair, nail and mucosal examination revealed normal results. Complete blood count (CBC), peripheral blood smear(PBS), serum urea and creatinine level did not deviate from normal findings. Ultrasonography of whole abdomen showed a mildly enlarged liver. A prominent lymph node in the aortocaval groove could be appreciated on the computed tomography(CT) scan of whole abdomen. CT scan of chest revealed bilateral axillary lymphadenopathy. Erythema necroticans, sarcoidosis and CTCL were considered as the differential diagnoses. Lesional biopsy revealed irregular epidermal hyperplasia, parakeratosis and epidermotropism in the absence of spongiosis.There was infiltration of atypical lymphocytes in the epidermis, forming well developed Pautrier’s micro abscesses. The lymphocytes were tagging the dermo-epidermal junction and within the epidermis showing surrounding halo, convoluted nuclei and variable nuclear pleomorphism. Band like papillary dermal lymphoid infiltrate, dermal lymphoid fibroplasia and nodular lymphoid aggregates could be appreciated in the deep dermis. Immunohistochemical tests were performed and the immunophenotypic profile revealed positivity for CD2, CD3, CD5, CD7, CD 8 and TIA1 and negative for CD4, CD20, CD30, CD56 and granzyme B. Based on clinical presentation, histopathology and Immunohistochemical tests, a final diagnosis of MF was made. The patient was then started on systemic antibiotic therapy, due to increased risk of bacterial super infection of lesions due to skin barrier disruption. He was then referred to Oncology department for further treatment. Conclusion: In this case, the male patient was suffering from mycosis fungoides. In the background of its clinical features being similar to other skin disorders like erythema necroticans, clinicians may face a diagnostic dilemma to correctly diagnose it. Timely diagnosis and treatment improves the prognosis in most cases. Histopathological evaluation still remains the investigation of choice.

Crystallization with Nodular Aggregation near the Glass Transition Temperature for Syndiotactic Polypropylene

The isothermal crystallization from the melt state of syndiotactic polypropylene (sPP) has been studied by wide-angle X-ray diffraction (WAXD), small-angle X-ray scattering (SAXS), and optical microscopy. The WAXD and SAXS results show the crystallization mechanism near the glass transition temperature in which the crystalline and mesomorphic nodules cover the entire sample with the formation of aggregation regions. For the SAXS analysis, the scattering function for the three-component system has been suggested. Furthermore, to analyze the growth kinetics of the aggregation region for sPP, the time-dependent structure factor combined with the homogeneous and inhomogeneous nucleation-and-growth kinetics has been suggested. The analysis shows that the growth kinetics of the aggregation region for sPP is the homogeneous nucleation-and-growth. The growth velocity of the aggregation region is a natural extrapolation of that of spherulite to the high supercooling region. These results might indicate that the crystallization with the nodular aggregation is a fundamental crystallization process near the glass transition temperature for polymers.

Tuberous Sclerosis Complex Associated Renal Cell Carcinoma with Leiomyomatous Stroma

This case report is about an unusual subtype of renal cell carcinoma with distinct morphological pattern in a 24-year-old female patient with tuberous sclerosis complex. She presented with pain in the left flank and an Magnetic Resonance Imaging (MRI) upper abdomen showed a solid cystic lesion in the interpolar region of left kidney, measuring 2.8×2.8×2.8 cm. She underwent partial nephrectomy which revealed a solid cystic tumour. Microscopically, the tumour was composed of long branching tubules lined by cells with voluminous clear/ vacuolated cytoplasm. A prominent fibromyomatous stroma was seen focally, separating the tumour into nodular aggregates. Tumour cells were positive for Cytokeratin 7 (CK7) on immunohistochemical study. These features were consistent with renal cell carcinoma with leiomyomatous stroma. Genetic study showed a heterozygous nonsense variation in Tuberous Sclerosis 1 (TSC1), gene, diagnostic of tuberous sclerosis. This report exemplifies the pivotal role pathologists play in the initial identification of certain hereditary cancer syndrome. The distinct morphology and immunohistochemical profile of the tumour is described.

S2509 Mantle Cell Lymphoma Presenting as Multiple Lymphomatous Polyposis: A Case Report

Introduction: Mantle Cell Lymphoma (MCL) typically presents with extensive lymphadenopathy, fevers, night sweats and unintentional weight loss. However, Multiple Lymphomatous Polyposis (MLP), MCL arising from the gastrointestinal tract, is an aggressive malignancy and is infrequently described. Case Description/Methods: A 68-year-old White man with atrial fibrillation and hypertension presented to the hospital with watery diarrhea, fatigue and intermittent epigastric pain for the past several weeks. He denied melena, hematochezia, heartburn, dysphagia, odynophagia, weight loss, loss of appetite, NSAID use, recent travel or sick contacts. There was no family history of gastrointestinal malignancy. No history of prior endoscopic evaluation. Physical exam was notable for brown stool on rectal exam. Further evaluation revealed Hgb 6.4, MCV 60.9, iron 43, TIBC 442 and iron saturation 10% consistent with iron deficiency anemia. Upper endoscopy was normal. Colonoscopy showed 8 6-16 mm polyps scattered throughout the colon, all of which were resected completely. Analysis showed a range of histology – 3 tubular adenomas, 1 serrated and 2 hyperplastic polyps. However, histology of 4 polypoid appearing lesions (Figure 1A) showed prominent nodular atypical submucosal lymphoid aggregates (Figure 1B) that were positive for CD20, cyclin D1, weak CD5, and negative for CD10 and CD23. This was consistent with the diagnosis of MCL. CT imaging showed diffuse lymphadenopathy, including bulky disease with a 16.6 x 10.6 cm small bowel mesenteric mass (Figure 1C), along with proximal small bowel and terminal ileal thickening. He was induced with 6 cycles of bendamustine-rituximab combination therapy and maintained on rituximab. A re-staging scan at 27 months showed no evidence of disease progression. Discussion: We present a case of MCL presenting as MLP with symptoms of anemia, diarrhea and epigastric pain. MCL is a rare B-cell non-Hodgkin's lymphoma that portends a poor prognosis, making early identification and diagnosis critical. Obstruction, GI bleeding, and perforation are common complications for MCL presenting as MLP. It is important to keep MLP on the differential when multiple small nodular or polypoid lesions are identified on colonoscopy. All different types of polyps should be resected or sampled during colonoscopy, and each evaluated by the pathologist to avoid missing clinically significant conditions. Early diagnosis is key to prevent morbidity and mortality in MLP as described in our case.Figure 1.: (A) polypoid lesion (B) nodular atypical submucosal lymphoid aggregates (C) 16.6 x 10.6 cm small bowel mesenteric mass.

Generalized nodules with the leonine appearance

A 39-year-old woman presented with generalized nodules and lymphadenopathy for 5 years, with nodules on the scalp coalesced to form larger cerebriform lesions. The nodules on the face and scalp gave a leonine appearance and some of them had erosions (Figs 1 and 2). The peripheral blood smear revealed atypical lymphocytes and eosinophils, whereas histopathology showed irregular nodular aggregates of intermediate and large lymphocytes, with vesicular nuclei and conspicuous nucleoli, histiocytes, and mononuclear cells in the dermis, extending up to the subcutaneous tissue.

IL-17RA receptor signaling contributes to lung inflammation and parasite burden during Toxocara canis infection in mice.

IL-17 is a cytokine produced by innate and acquired immunity cells that have an action against fungi and bacteria. However, its action in helminth infections is unclear, including in Toxocara canis infection. Toxocariasis is a neglected zoonosis representing a significant public health problem with an estimated seroprevalence of 19% worldwide. In the present study, we describe the immunopathological action of IL-17RA in acute T. canis infection. C57BL/6j (WT) and IL-17RA receptor knockout (IL-17RA-/-) mice were infected with 1000 T. canis eggs. Mice were evaluated 3 days post-infection for parasite load and white blood cell count. Lung tissue was harvested for histopathology and cytokine expression. In addition, we performed multiparametric flow cytometry in the BAL and peripheral blood, evaluating phenotypic and functional changes in myeloid and lymphoid populations. We showed that IL-17RA is essential to control larvae load in the lung; however, IL-17RA contributed to pulmonary inflammation, inducing inflammatory nodular aggregates formation and presented higher pulmonary IL-6 levels. The absence of IL-17RA was associated with a higher frequency of neutrophils as a source of IL-4 in BAL, while in the presence of IL-17RA, mice display a higher frequency of alveolar macrophages expressing the same cytokine. Taken together, this study indicates that neutrophils may be an important source of IL-4 in the lungs during T. canis infection. Furthermore, IL-17/IL-17RA axis is important to control parasite load, however, its presence triggers lung inflammation that can lead to tissue damage.

Unique Isospora-associated histologic lesions in white-rumped shama (Copsychus malabaricus).

Twenty-one white-rumped shamas (19 necropsied, 2 biopsied) (Copsychus malabaricus) housed at the San Diego Zoo between 1992 and 2020 were diagnosed with Isospora infection based on evaluation of histologic sections. Review of these cases revealed a consistent histologic lesion characterized by nodular aggregates of atypical epithelioid macrophages containing few intracytoplasmic protozoa, with or without lymphocytic infiltrates. Of the 19 necropsied cases, 16 (84%) had systemic lesions variably affecting the liver, spleen, gastrointestinal tract, lung, pancreas, connective tissues, or bone marrow, while all 21 diagnosed cases had skin involvement. The findings suggest that white-rumped shamas have a unique inflammatory response to isosporosis with a predilection for the skin. Skin may be a diagnostically sensitive sampling site for histologic diagnosis of Isospora in this species.

Limited sinonasal Rosai-Dorfman disease presenting as chronic sinusitis.

AimsThe sinonasal tract is a common extranodal site for Rosai–Dorfman disease (RDD). Recently, histiocytes with features of RDD were identified in the clinical setting of chronic sinusitis. This study evaluates whether this phenomenon should be considered part of the RDD spectrum or classified separately as RDD‐like histiocytes.Methods and resultsWe prospectively collected 13 cases showing histological features of RDD in chronic sinusitis patients and identified 14 with similar findings (3.5%) via retrospective review of 403 sinus contents over 2 years. All 27 cases displayed nodular aggregates of eosinophilic histiocytes with intermixed lymphoplasmacytic inflammation, prominent eosinophils and emperipolesis. The histiocytes were positive for S100 protein and cyclin D1 and negative for CD1a and CD207. All patients presented with severe chronic sinusitis without tumour formation or systemic symptoms. Twelve patients with follow‐up (55%) required repeat sinus surgery compared with just 43 other sinusitis patients evaluated (11%); features of RDD were present in their additional specimens. Two cases that underwent targeted next‐generation sequencing (20%) had oncogenic mutations in NF1 and KEAP1.ConclusionsOverall, these findings confirm diagnostic histological and immunohistochemical features of RDD in a subset of chronic sinusitis specimens. While patients uniformly lack systemic involvement or tumefactive growth, they have a high risk of recurrent sinus disease. Although the relatively subtle nature of the findings raises consideration of separate classification, the presence of occasional oncogenic mutations and evidence of consistent MAPK/ERK pathway activation via cyclin D1 positivity suggests that this phenomenon represents a unique limited manifestation of RDD.

Uterine Adenomyoma- A Case Report

Abstract: Adenomyoma of the uterus is a benign tumour composed of benign endometrial glands and endometrial stroma bordered by leiomyomatus smooth muscle. It is classified as a mixed epithelial and mesenchymal tumour of uterine corpus. It may be located within the myometrium, or it may involve or originate in th endometrium and grow as a polyp. We report a case of uterine adenomyoma that occurred in 32 year old women. The patient presented with menorrhagia and irregular periods and she underwent a hysterectomy with bilateral salphingo-oopherectomy. Histologically, tumour was composed of nodular aggregate of irregular and cystically dilated endometrial glands surrounded by fibromyomatous stroma and were diagnosed as adenomyoma of uterus.

Clinically Undetected Hodgkin Lymphoma Diagnosed Initially on Bone Marrow Biopsy: A Large Retrospective Observational Study from a Tertiary Care Center

Abstract Introduction Hodgkin lymphoma (HL) involving the bone marrow (BM) is relatively rare with an incidence ranging from 4% to 18%. The incidence of primary HL of marrow is 0.25%. To the best of our knowledge, the present study is the largest study on HL diagnosed initially on marrow biopsy. Objective To establish diagnostic criteria based on clinicopathological and histological features in HL diagnosed first on the marrow. Materials and Methods This was a retrospective study done from January 2012 to December 2020 that included 36 cases of HL diagnosed initially on BM. Based on the presence of large mononuclear or binucleate Reed–Sternberg (RS)-like cells in a polymorphous inflammatory background, HL was suspected and immunohistochemistry (IHC) with CD15 and CD30 was done. Correlation with subsequent lymph node biopsies was done, wherever possible. Results Fever (94.4%) was the most common symptom, followed by loss of weight (66.7%). Twenty-one cases (58.4%) had uni/bicytopenia and 15 cases (41.6%) had pancytopenia. Only one case showed suspicious mononuclear RS cells on aspirates and the rest of the cases were diagnosed on trephine biopsy alone. Trephine imprints showed variable cellularity in 13 (36%) cases. Diffuse involvement was seen in 24 cases (66.7%), and focal nodular aggregates were seen in 12 cases (33.3%). Out of 36 cases, 26 cases (19 cases on marrow and 7 cases on lymph node) were confirmed as HL with IHC. Immunophenotype of the RS cells on the marrow was CD30+/CD15+ in (6/29) (20.7%) cases, CD30+/CD15− in (7/29) (24.1%) cases and CD30−/CD15+ in (6/29) (20.7%) cases. Seven cases (26.9%) were diagnosed on subsequent lymph node biopsy as mixed cellularity HL with IHC confirmation. Marrow fibrosis was seen in 16 cases (44.4%), and granulomas were seen in 8 cases (22.2%). Conclusion In cases presenting with long-standing fever and cytopenias, HL must always be suspected, even if there are no palpable lymph nodes. Bone marrow biopsy is preferable over aspiration in such cases and IHC plays a major role in diagnosing the cases.

A Pigeon-Derived Sub-Genotype XXI.1.2 Newcastle Disease Virus from Bangladesh Induces High Mortality in Chickens.

Newcastle disease virus (NDV) is a significant pathogen of poultry; however, variants also affect other species, including pigeons. While NDV is endemic in Bangladesh, and poultry isolates have been recently characterized, information about viruses infecting pigeons is limited. Worldwide, pigeon-derived isolates are commonly of low to moderate virulence for chickens. Here, we studied a pigeon-derived NDV isolated in Bangladesh in 2010. To molecularly characterize the isolate, we sequenced its complete fusion gene and performed a comprehensive phylogenetic analysis. We further studied the biological properties of the virus by estimating mean death time (MDT) and by experimentally infecting 5-week-old naïve Sonali chickens. The studied virus clustered in sub-genotype XXI.1.2 with NDV from pigeons from Pakistan isolated during 2014–2018. Deduced amino acid sequence analysis showed a polybasic fusion protein cleavage site motif, typical for virulent NDV. The performed in vivo pathogenicity testing showed a MDT of 40.8 h, and along with previously established intracerebral pathogenicity index of 1.51, these indicated a velogenic pathotype for chickens, which is not typical for pigeon-derived viruses. The experimental infection of chickens resulted in marked neurological signs and high mortality starting at 7 days post infection (dpi). Mild congestion in the thymus and necrosis in the spleen were observed at an advanced stage of infection. Microscopically, lymphoid depletion in the thymus, spleen, and bursa of Fabricius were found at 5 dpi, which progressed to severe in the following days. Mild to moderate proliferation of glial cells was noticed in the brain starting at 2 dpi, which gradually progressed with time, leading to focal nodular aggregation. This study reports the velogenic nature for domestic chickens of a pigeon-derived NDV isolate of sub-genotype XXI.1.2. Our findings show that not all pigeon-derived viruses are of low virulence for chickens and highlight the importance of biologically evaluating the pathogenicity of NDV isolated from pigeons.

Reassessment of EWSR1 gene rearrangement-negative undifferentiated round cell sarcomas in bone and soft tissues by fluorescence in situ hybridization

Objective: To unravel the CIC rearrangement sarcomas and BCOR-CCNB3 sarcomas from EWSR1 rearrangement-negative undifferentiated round cell sarcomas in the bone and soft tissues. Methods: Twenty-eight cases of EWSR1 rearrangement-negative undifferentiated round cell sarcomas of bone and soft tissues, tested for CIC rearrangement and BCOR rearrangement by fluorescence in situ hybridization and related immunostaining were analyzed, and some of the BCOR rearrangement cases were verified by reverse transcription-polymerase chain reaction. Results: Five of 28 (17.9%) tested cases were positive for CIC rearrangement and six (21.4%) for BCOR rearrangement. Histopathologically, CIC rearrangement sarcomas comprised nodular aggregates of round to polygonal cells, containing hyperchromatic nuclei, prominent nucleoli and moderate cytoplasm, with focal variable necrosis and myxoid stroma. BCOR-CCNB3 sarcomas mostly comprised diffusely arranged, round to oval to short spindly cells with angulated nuclei, vesicular chromatin, inconspicuous nucleoli and interspersed vessels. Immunohistochemically, five of six BCOR-CCNB3 sarcomas showed CCNB3 immunostaining, which could be helpful for diagnosis. Two patients with CIC rearrangement sarcoma died of the diseases in seven months and twenty-two months. One patient with BCOR-CCNB3 sarcoma died of the diseases in forty-six months. Conclusions: Overall, 39.3% of the EWSR1 rearrangement-negative undifferentiated round cell sarcomas are CIC rearrangement sarcomas and BCOR-CCNB3 sarcomas. Molecular testing is helpful for diagnosis.

Different Conformations of Fibronectin Fibrils Formed Under Cell-Free Condition in Vitro display Distinct Effects on Cell Adhesion and Spreading

Introduction: Insoluble fibrils are the active form of fibronectin (FN) with exposed functional domains capable of interacting with a variety of biomolecules and cells to support many cellular processes. Recent studies show that the formation of FN fibrils can occur in cell-free systems. Currently, the characteristic of FN fibrils and their potential in wound healing application are not fully understood. Our project aims at (i) using urea to induce the formation of FN fibrils with different conformations and (ii) to characterize their conformation-dependent effect on cellular adhesion and spreading.Method: To induce FN fibrillogenesis, native plasma FN at various initial concentrations (0.25, 0.5, 0.75 and 1 mg/ml) was treated by dialysis in urea at concentrations of 1, 2, and 4 M for 16h at room temperature followed by dialysis in PBS buffer (to remove urea). Morphological analysis of formed FN fibrils was conducted bylight microscopy and scanning electron microscopy (SEM). To assess biological effects of FN fibrils on cells in relationto their morphologies, platelet and fibroblast cell adhesion assays were performed. Specifically, platelet rich plasma (5x107 platelets/ml) or fibroblasts (L929, 104 cells/ml) were incubated on surfaces pre-coated with 0.25 and 0.75 mg/ml native plasma FN or FN fibrils for 30 min at 370C.Results: Our cell-free FN fibrillogenesis experiments in vitro indicated that the optimal concentrations of FN and urea were 1 mg/ml and 2 M, respectively, to induce formation of FN fibrils with various conformations. FN only formed minuscule fibrils at 1 M urea while there was no fibril formation at 4 M urea. In contrast, FN fibrils with various conformations ranging from fibrillar FN to non-fibrillar FN (i.e., nodular FN aggregates) were formed when using 2 M urea. Fibril formation was shown to be dependent on the FN concentration. FN at low concentrations (0.25 or 0.5 mg/ml) formed a few insoluble minuscule fibrils or nodular FN aggregates with variation in size ranging from 10 to 50 µm. FN at higher concentrations (0.75 or 1 mg/ml) formed much more nodular FN and fibrillar FN with diameters mostly ranging from 10 to 30 µm that tended to interlace into the fibrillar matrix. SEM analysis documented morphological variations of fibrils formed under different conditions. For example, at low FN concentration (0.25 mg/ml) small fibrils with a smooth surface were formed. By contrast, larger fibrils with attached nodular FN and a resulting rough surface were formed at high FN concentration (≥ 0.75 mg/ml). Adhesion assays indicated that platelets adhered approximately 6 times more firmly on FN fibrils than on native plasma FN (n=3, p < 0.05).On immobilized native FN, adherent platelets were evenly distributed or formed small clusters, while fibrillar FN induced platelet accumulation on the matrix. Fibroblasts adhered both on non-fibrillar FN and matrix of small fibrils (diameter in range of 10-30 µm) but did not interact to larger fibrillar bundles (diameter > 292 µm). Investigations of FN-cell interaction by SEM revealed a structure-dependent effect of FN on fibroblast morphology after adhesion. Specifically, cells remained in spherical shape and separately adhered on nodular FN while extending their shape in multiple directions on the rough surface of FN fibrils.Conclusion: Treatment of native plasma FN by urea at a concentration of 2 M induces irreversible formation of heterogeneous FN fibrils, displaying a rough surface due to attachment of FN nodules. The conformational and compositional changes of the FN matrix can lead to firm adhesion of platelets and fibroblasts and may thus provide a future tool to improve wound healing upon local application. DisclosuresNo relevant conflicts of interest to declare.

AYURVEDIC MANAGEMENT OF ADENOMYOSIS – A CASE REPORT

Most of the women are suffering from one or other menstrual problem in their life, Adenomyosis is also one among them. In general, Adenomyosis is defined as presence of endometrial tissue within the myometrium. It is described as circumscribed nodular aggregate of smooth muscle and endometrial glands with compensatory hypertrophy of the myometrium surrounding the ectopic endometrium1. It is associated with menorrhagia or menometrorrhagia and congestive dysmenorrhoea. In Ayurvedic text 20 yoni Vyapads are explained, among them Vipluta is one of them characterised with constant pain associated with other pains of Vata Prakopa2. So, in this case we diagnosed the case as Vipluta Yonivyapad and treated with the same line of treatment, which got promising result. Keywords: Adenomyosis, Yoni Vyapad, Vipluta Yoni Vyapad.