Pleomorphic Cells Research Articles

First Report of a ‘Candidatus Phytoplasma aurantifolia’ Strain Associated with Virescence, Floral Proliferation, and Dwarf Symptoms on Indigofera suffruticosa in Taiwan

Indigofera suffruticosa Mill., generally known as anil or Guatemalan indigo, is a perennial shrub planting in tropical and subtropical regions, including Taiwan; it was commonly used to produce the indigo dye, which is still being used in many personal workshops in Taiwan. There were a few reported cases of the association of phytoplasma with Indigofera spp. In 1983, I. suffruticosa showing phyllody and small leaf symptoms was first described, but no molecular identification was conducted (Albanese et al. 1995). Indigofera linifolia with little leaf, phyllody, and proliferation symptoms and Indigofera colutea showing little leaf symptoms in Australia caused by phytoplasma strains were reportedly associated with papaya yellow crinkle disease (Padovan and Gibb 2001). Affected Indigofera juncea and Indigofera hirsuta exhibiting witches’-broom symptoms were reported to be the same as the sesame phyllody phytoplasma isolate in Thailand (Nakashima et al. 1995). In February 2016, I. suffruticosa plants showing virescence, floral proliferation, and dwarf symptoms were collected from commercial fields of Mingjian Township, Nantou County, Taiwan. Two each of symptomatic and asymptomatic samples were collected from the field for further study. Transmission electron microscopic observation revealed the presence of the pleomorphic phytoplasma cells only in the phloem sieve cells of the two symptomatic I. suffruticosa plants. The two asymptomatic and two symptomatic samples were used for total DNA extraction with the modified CTAB method, and the amplicons of the nested PCR with universal primer pairs for nearly full-length 16S rDNA amplification (P1/P7 followed by R16F2n/R16R2) were electrophoresed in 0.8% agarose gel (Fulton et al. 1995; Lee et al. 1998). A 1.2-kb fragment (GenBank accession no. KX268346) was identified only from the two symptomatic I. suffruticosa plants. Analysis of 16S rDNA sequences by iPhyClassifier indicated that the phytoplasma associated with virescence symptoms in I. suffruticosa shared a 98.2% identity with that of the ‘Candidatus Phytoplasma aurantifolia’ reference strain (GenBank accession no. U15442). Virtual restriction fragment length polymorphism patterns were identical to those of the 16SrII-V subgroup (Zhao et al. 2009). Taken together, the results indicated that the phytoplasma associated with I. suffruticosa in Taiwan is a ‘Ca. P. aurantifolia’-related strain. The occurrence of phytoplasma in I. suffruticosa could have direct implication on economic profit for growers and the handicraft industry in Taiwan. To our knowledge, this is the first report of a 16SrII-V subgroup phytoplasma causing virescence, floral proliferation, and dwarf symptoms on I. suffruticosa in Taiwan.

Metastatic pleomorphic dermal sarcoma to small bowel – A case report

Pleomorphic dermal sarcoma (PDS), a dermal based tumour that is related to atypical fibroxanthoma with overlapping morphological and immunohistochemical features but with subcutaneous invasion, necrosis, perineural or vascular invasion. PDS has low malignant potential with risk of local recurrence and metastases. Reported sites of metastases are distant skin, lymph nodes and lung. We report a case of a male aged 65 who presented with gastrointestinal bleeding due to haemorrhage from a small bowel lesion. His medical history included Muir-Torre syndrome, stage IV colon cancer (with liver metastases) and PDS of the lower lip (resected February 2018) with known lung metastases. He underwent a segmental resection of small bowel in June 2019; macroscopically a solitary 39 mm lesion was noted. Histopathology showed a tumour composed of sheets and nests of pleomorphic, mitotically active, epithelioid and spindle cells. Immunohistochemically the malignant cells showed positive for CD10 and negative staining for cytokeratins, CD117 and melanoma markers. The morphology and immunophenotype were the same as his previously diagnosed lip PDS. This is the first reported case of metastatic PDS to small bowel, and highlights the importance of clinical-pathological correlation. Although rare, metastatic sarcoma should be considered in the differential of pleomorphic epithelioid/spindled small bowel neoplasms.

Hypereosinophilia in a patient with refractory eczema and ALK-negative anaplastic large cell lymphoma

A 41-year-old man with refractory eczema on immunosuppressants developed swinging fever and dyspnoea, with left pleural effusion and left axillary and groin lymphadenopathy (2–3 cm) found in physical examination. Clinically there was no eczema flare-up. Laboratory tests showed hypereosinophilia (79.1×109/L) and elevated LDH (520 U/L) and ALP (187 U/L). Some dysplastic eosinophils showing hyposegmented nuclei and hypogranular cytoplasm were noted on blood smears. Workup for parasites, allergy and autoimmune markers was unremarkable. Thoracentesis yielded exudative effusate with numerous lymphocytes (WBC >1000/mm3) but no malignant cell was evident. FNAC of lymph nodes found eosinophilic infiltrate only. Bone marrow examination revealed marked eosinophilia with similar dysplastic features and megakaryocytic hyperplasia, raising suspicion of myeloproliferative neoplasms. Karyotyping, molecular tests for JAK2 mutation and BCR-ABL1 fusion, and FISH for FIP1L1-PDGFRA, PDGRFB and FGFR1 rearrangements were unremarkable. Excisional biopsy of left groin lymph node was then pursued and demonstrated sheets of pleomorphic neoplastic cells with focal ‘hallmark cells’. Anaplastic large cell lymphoma (ALK–CD30+CD3+CD8+) was diagnosed. PET/CT scan revealed a stage-IV disease with extensive supra- and infra-diaphragmatic hypermetabolic lymph nodes and multiple bone involvements. The patient opted for brentuximab vedotin plus combination chemotherapy. This case underscores the importance of exhaustive investigations and clinicopathological corroboration in the workup of hypereosinophilia.

Proliferating cell nuclear antigen antibody testing – Getting them all

Proliferating cell nuclear antigen antibody (PCNA) is associated with systemic lupus erythematosus (SLE) and characterises a cohort of patients at risk of renal or central nervous system involvement. The antibody has also been reported in patients with hepatitis B and C virus infection. It has a pleomorphic nuclear cell cycle dependant pattern on antinuclear antibody (ANA) testing by indirect immunofluorescence (IFA). In this laboratory PCNA has been historically reported from the IFA pattern on Hep2000 cells (Immuno Concepts, USA) and not confirmed on a secondary test. An audit of PCNA pattern reported from ANA testing and incidental characterisation by immunoblot was undertaken to determine the frequency of PCNA detected. Over 4 years, 34 PCNA results were reported. Clinical details provided included SLE, joint pain and acute hepatitis. Extractable nuclear antigen antibody characterisation by immunoblot incidentally identified a further 20 samples with PCNA not reported on ANA. ANA patterns included speckled, homogeneous and nucleolar. Clinical details in this group also included SLE and joint pain. This investigation demonstrates that IFA testing alone for PCNA may miss clinically relevant cases due to a strong dominant ANA pattern. Clinical characteristics and correlation with IFA is required to validate the immunoblot PCNA result.

A Comparison of the Pathological Types of Undifferentiated Carcinoma of the Pancreas

This study aimed to identify the detailed clinicopathological features of undifferentiated carcinoma of the pancreas (UCP). We investigated clinical, imaging features and the prognoses of 261 patients; 8 were our patients, and the remainder were identified by searching English-language articles in PubMed. We classified patients with UCP into 3 types based on pathological findings: osteoclast-like giant cell-associated carcinoma, pleomorphic cell carcinoma (PLC), and spindle cell carcinoma. There were no remarkable differences in clinical, radiological features between these 3 types. However, PLCs were significantly more likely to be unresectable than were the other 2 types (P < 0.001). Patients with osteoclast-like giant cell-associated carcinoma achieved the best overall survival (OS) rates (P < 0.001), whereas those with spindle cell carcinoma had significantly longer OS rates than did those with PLC (P = 0.004). These OS patterns were maintained when considering only those patients who underwent resection. Patients with PLC had both lower curative resection and high lymph node metastasis rates (P = 0.029, P = 0.023). Patients who underwent resection had more favorable prognoses than did those who did not. Surgery is the first choice for resectable UCP. Pleomorphic cell carcinoma is particularly malignant; postoperative treatment should be introduced immediately.

52. FGFR3-TACC3: Case report and review of a potentially targetable diagnostic marker in high grade gliomas

Background: FGFR3-TACC3 fusion has been identified in IDH wild-type WHO grade III and IV astrocytomas, with an incidence of 2–3% in most case series and a range of 1–8%. These tumours have characteristic histopathological features and a poor prognosis. The aberrant CD34 immunohistochemical staining and glial and neuronal marker co-expression may lead to a misdiagnosis of a glioneuronal tumour with a better prognosis. An immunohistochemistry antibody to FGFR3 is commercially available which may serve as a more cost-effective screening test for identification of possible FGFR3-TACC3 cases. These tumours may also be amenable to treatment by FGFR tyrosine kinase inhibitors, or through inhibition of oxidative phosphorylation as a down-stream functional target of FGFR3-TACC3 fusion. Aims: A report highlighting common pitfalls in the diagnosis of high grade astrocytoma with FGFR3-TACC3 gene fusion. Methods: A single case study and review of the supporting literature. Results and conclusions: A 64-year-old male with a four month history of focal sensory abnormalities had a solitary 24 mm, intra-axial, contrast-enhancing left parietal lobe brain lesion on MRI. A stereotactic craniotomy and tumour resection was performed. Histopathology showed a malignant heterogeneous tumour. The cellular morphology included gemistocytic features, larger pleomorphic cells and numerous mitotic figures. Additional findings were psammomatous calcifications and thin-walled branching blood vessels with perivascular lymphoid cuffing. Many of the lesional cells co-expressed neuronal and glial protein markers on immunohistochemistry and there was also patchy aberrant CD34 staining. IDH1, IDH2 and BRAF genes were wild-type on molecular pathology testing. There was no loss of chromosome 1p or 19q or EGFR amplification by cytogenetic analysis. The Illumina TP170 next-generation sequencing assay identified a FGFR3-TACC3 gene fusion, a TERT promoter mutation (C250T) and a gain of FGFR3. The diagnosis of anaplastic astrocytoma, WHO grade III, IDH wild-type with FGFR3-TACC3 fusion, TERT promoter mutation and FGFR3 gain was made.

Cutaneous pilomatrical carcinosarcoma: a case report with molecular analysis and literature review

BackgroundCutaneous pilomatrical carcinosarcoma (CS) is a very rare biphasic tumor composed of admixed epithelial and mesenchymal malignant cells, with limited information on its pathogenesis. We report a case of pilomatrical CS of the scalp with comparative immunohistochemical and molecular analysis together with a review of the literature.Case presentationA 74-year-old woman presented with a rapidly growing long-standing tumor of the scalp. The tumor was surgically resected. Histologically, the tumor was 25 mm in diameter, and was composed of carcinoma showing a clear pilomatrical differentiation and sarcoma with pleomorphic spindle cells and giant cells. Both epithelial and mesenchymal components shared focal cytoplasmic and/or nuclear accumulation of β-catenin based on immunohistochemical analysis, although a mutation of exon 3 of the CTNNB1 gene was not detected. Fluorescence in situ hybridization analysis revealed gains of chromosomes 9p21, 3, and 7 in both the epithelial and sarcomatous components.ConclusionsThe current case demonstrated characteristic findings of pilomatricoma and further evidence of partial clonality between the carcinomatous and sarcomatous component, suggesting the possibility of malignant transformation of pilomatricoma. Rapid growth of a pilomatrical tumor should warrant the development of a malignant tumor, including CS.

Cytological and Histopathological Correlation of Feline Giant Cell-Rich Osteosarcoma

Background: Primary bone tumours are uncommon and poorly reported in cats but osteosarcoma (OSA) is the most frequent, mostly in elderly animals. Giant cell-rich OSA is considered rare in the literature representing 3% of all OSA in humans. The mitotic index seems to have a significant effect on the survival time of cats affected by this neoplasm as well as the tumour histopathological grade. The objective of this study was to report the cytological and histopathological findings of a giant cell rich OSA in a 4-year old cat with persistent feline leukemia virus (FeLV) antigenaemia.Case: A 4-year-old male neutered cat was referred with a history of persistent FeLV viraemia and pelvic limb lameness with a firm swelling. Previous radiographs of the affected limb revealed bone lysis in the third and fourth metatarsals and increased soft tissue radiopacity in the tarsal region. The referral veterinary assumed it to be osteomyelitis and initiated clinical treatment with antibiotic and anti-inflammatory. The cat was referred after there was no response to medical treatment. The cat was presented with a 5cm diameter ulcerated mass, with putrid odor in the pelvic limb. Complementary exams were performed, and abnormalities were found, including increased urea, creatinine, calcium and potassium, and decreased sodium and phosphorus. A new radiograph showed exuberant bone proliferation, with increased radiopacity involving tarsal, metatarsal, distal third of tarsal I and II, and distal diaphysis of metatarsal V, without compromising the metaphyseal region of distal diaphysis of metatarsal IV. Chest radiographs and abdominal ultrasound were unremarkable. Fine-needle aspiration was performed for cytological analysis, which reavealed a moderate amount of pleomorphic mesenchymal cells with moderate adhesion, cytoplasm with a format ranging from fusiform to stellate, pronounced anisocytosis and cellular pleomorphism, and elevated nucleus:cytoplasm ratio. Nucleus was oval and presented loose chromatin, single to double large and evident nucleolus, frequent karyomegaly, along with marked anisocariosis and nuclear pleomorphism. Multinucleated giant cells were and there was a single mitotic figure in 12 high power fields (0.196 mm2 FN20/400x). Therefore, it was suggestive of malignant mesenchymal neoplasia with possibility of OSA, fibrosarcoma oe undifferentiated sarcoma. Limb amputation with femoral disarticulation was performed uneventfully. The material was conserved in 10% formalin, submitted to macroscopic and microscopic evaluation, which showed a large number of fusiform and stellate cells, with indistinct edges, scarce eosinophilic cytoplasm, high nucleus:cytoplasm ratio and oval nucleus, presenting moderate to marked anisocariosis, loose chromatin, with unique and evident nucleoli, besides of bone trabeculae. Nuclear pleomorphism was moderate and there was four mitotic figures in three random high-power fields (400x). It was observed areas of vascular ectasia, and neoplastic embolization in lymphatic and blood vessels. Among the cells, collagenous stroma was predominant but in some areas there was an eosinophilic amorphous material with the possibility of osteoid matrix or collagen. There was also a large numbers of multinucleated giant cells. The histopathological result was compatible with a grade III giant cell rich OSA.Discussion: Although bone tumours are uncommon in cats, OSA is the most frequent, affecting maingly middle-aged to elderly cats, with a mean age of 10 years, which is different from the present report in a 4-year old cat, with FeLV persistent viraemia. Retroviral status may have influenced the development of the disease at na early onset. FeLV induces uncontrolled cell proliferation through insertional mutagenesis (usually near myc) inducing malignant neoplasias, mainly lymphoma, but also multiple cartilaginous exostosis, which, along with osteomyelitis and bone cyst were included in this patient´s list of differential diagnoses. Despite the macroscopic and radiographical andagressiveness there was no metastasis identified through chest radiographs or abdominal ultrasound, and feline OSA is associated with a lower metastatic rate, if compared to canine OSA. Cytological analysis was compatible with malignant mesenchymal neoplasia, being suggestive of giant cell rich OSA. There is no cytological classification for OSA, however cytological findings of malignancy may be correlated with the patient's clinical course. Cytological features were in agreement with the histopathological findings, compatible with a grade III giant cell rich OSA. Therefore, it is concluded that the characteristics of malignancy presented by cytology were sufficient for the recommendation of amputation of the affected limb, once there was no imaging sign of metastases in the chest or abdomen.

Multiple Myeloma in a Feline

Background: Multiple myelomas (MM) are malignant neoplasms originating in the bone marrow plasmacytes, and are characterized by the presence of persistent hyperglobulinemia. Although they are rarely found in domestic animals, the canine species is most affected. In felines, they represent less than 1% of hematopoietic neoplasms, and mainly occur in older animals aged 10-12 years. The aim of the present study was to report a case of multiple myeloma, with extramedullary plasmocytosis in a feline.Case: A 3-year-old feline, female, mixed breed, with a history of apathy, anorexia, halitosis, and vomiting was referred to the Laboratório Regional de Diagnóstico (LRD) at the Universidade Federal de Pelotas (UFPel). Oral examination revealed a cyanotic oral mucosa and ulcerative lesions on the gingiva. The necropsy showed generalized lymphadenomegaly, splenomegaly, and hemorrhagic diathesis. Organ fragments were collected and fixed in 10% buffered formalin. After 48 h, the samples were cleaved, embedded in paraffin, and stained using the routine hematoxylin and eosin technique. Selected sections were subjected to immunohistochemistry (IHC) using the streptavidin-biotin method. The CD79a, CD138, CD3, and MUM1 antibodies were used. Histopathological evaluation of the bone marrow revealed hypercellularity with predominant round pleomorphic cells (90%), which presented an eccentric basophilic nucleus and abundant eosinophilic cytoplasm. The same neoplastic cells infiltrated the gingival mucosa, lymph nodes, spleen, liver, kidneys, heart, lungs, and brain. The IHC showed positive immunostaining for MUM1 and CD138 antibodies.Discussion: The diagnosis of MM in the present study was based on histopathology and by IHC. According to the literature, the presence of neoplastic plasmocytes in the bone marrow (&gt;20%), osteolytic lesions, monoclonal gammopathy, and Bence-Jones proteinuria should be investigated in suspected MM cases. The presence of two or more of these characteristics is necessary for confirmed diagnosis. However, studies show that only 50% of felines with MM have two or more of these characteristics, with cell morphology and neoplastic infiltration in other organs being important parameters for the definitive diagnosis of MM in them. For MM diagnosis by IHC, CD79a, CD138, and MUM1 can be used. In this report, there was positive immunostaining for MUM1 and CD138, which allowed the exclusion of other neoplasms of hematopoietic origins. The origin of MM is unknown, but chronic immune stimulation is believed to be involved in humans. In felines, although infection by the feline immunodeficiency virus (FIV) and feline leukemia (FeLV) is commonly associated with the occurrence of lymphomas, the same is not described for MM. This was corroborated in the present study, since the animal in this report tested negative for FIV and FeLV. In cats, MM has no apparent predilection for race or sex, and mainly affects older animals (average age, 10 years). Although MM is rare, it has been described not only in this case, but also in young cats. The infiltration of neoplastic plasmacytes in the extramedullary tissues (extramedullary plasmacytosis) is observed more frequently in felines than in canines, and mainly occurs in the lymph nodes, spleen, and liver. In this case, there was disseminated extramedullary plasmacytosis involving multiple organs, such as the brain, lungs, heart, and kidneys. Persistent hyperglobulinemia is the main clinical manifestation of MM observed in 87.7% of cats with this neoplasm. However, this variation can also be observed in other pathologies, such as chronic lymphocytic leukemia, B cell lymphoma, primary macroglobulinemia, and chronic infections (e.g., ehrlichiosis, leishmaniasis, and heartworm disease). Hence, MM should always be considered as differential diagnosis when diagnosing the aforementioned diseases.

Soft Tissue Special Issue: Cutaneous Pleomorphic Spindle Cell Tumors.

This manuscript provides an overview of pleomorphic spindle cell tumors presenting on sun-damaged skin of the elderly and includes discussions of atypical fibroxanthoma, pleomorphic dermal sarcoma, spindle cell and metaplastic squamous cell carcinoma, spindle cell and dedifferentiated melanoma and poorly differentiated cutaneous angiosarcoma. These tumors share many of the clinical presenting and histological features, making confident diagnosis challenging. A reliable and robust diagnosis is necessary to predict behavior as the biologic potential of these tumors ranges from benign (e.g. atypical fibroxanthoma) to outright malignant with poor survival rates (e.g. cutaneous angiosarcoma). The salient clinical, histologic and immunohistochemical characteristics are discussed in detail with emphasis on distinguishing features and differential diagnosis to provide the reader with a better understanding of these entities and helpful clues for a more robust diagnosis.

Practical Approach to the Histologic Diagnosis of Gastrointestinal Lymphomas Through the First-line Marker Battery of CD20, CD3, CD30, and Epstein-Barr Virus-encoded RNAs.

The gastrointestinal (GI) tract is a prevalent site for extranodal lymphomas. Some subtypes of GI tract lymphomas are aggressive and have dismal clinical outcomes. Therefore, prompt histopathologic detection of such types can be very important. We thus introduce a practical approach in the histopathologic diagnosis of GI lymphomas according to the revised World Health Organization (WHO) classification. When lymphocyte proliferation is found in the GI tract, a stepwise approach can help narrow down the differential diagnoses. When considering subtype incidence, macroscopic findings, and microscopic patterns, applying a first-line marker battery of CD20, CD3, CD30, and Epstein-Barr virus-encoded RNAs can effectively narrow down the top differential diagnoses at the initial step. Generally, the most common subtype among GI tract lymphomas is B-cell non-Hodgkin lymphoma identified by CD20 expression, followed by T-cell and NK-cell non-Hodgkin lymphomas identified by the CD3 expression, and some subtypes are defined by Epstein-Barr virus infection or CD30 expression. Macroscopically, lymphomas present as various gross types, such as large masses, small lesions, superficial and shallow lesions, polyp-like or polyposis-like features, or ulcer/necrosis/perforation. Microscopically, large pleomorphic cells or small to medium-sized tumor cells grow with various architectures and tumor microenvironments. Incorporation of macroscopic and microscopic features and the stepwise immunophenotyping may be a practical approach to the differential diagnosis of aggressive lymphoma, indolent/low-grade lymphoma, or benign to indolent lymphoproliferative disease. Exceptions always exist; this approach focuses on the relatively prevalent circumstances of lymphomatous lesions initially encountered in the GI tract.

In vitro evaluation of electroporated gold nanoparticles and extremely-low frequency electromagnetic field anticancer activity against Hep-2 laryngeal cancer cells.

The extremely-low frequency electromagnetic field (ELFEMF) has been proposed for use in cancer therapy since it was found that magnetic waves interfere with many biological processes. Gold nanoparticles (Au-NPs) have been widely used for drug delivery during cancer in vitro studies due to their low cytotoxity and high biocompatibility. The electroporation of cancer cells in a presence of Au-NPs (EP Au-NPs) can induce cell apoptosis, alterations of cell cycle profile and morphological changes. The impact of ELFEMF and EP Au-NPs on morphology, cell cycle and activation of apoptosis-associated genes on Hep-2 laryngeal cancer cell line has not been studied yet. ELFEMF on Hep-2 cells were carried out using four different conditions: 25/50 mT at 15/30 min, while Au-NPs were used as direct contact (DC) or with electroporation (EP, 10 pulses at 200V, equal time intervals of 4 sec). MTT assay was used to check the toxicity of DC Au-NPs. Expression of CASP3, P53, BAX and BCL2 genes was quantified using qPCR. Cell cycle was analyzed by flow cytometry. Hematoxylin and eosin (HE) staining was used to observe cell morphology. Calculated IC50 of DC Au-NPs 24.36 μM (4.79 μg/ml) and such concentration was used for further DC and EP AuNPs experiments. The up-regulation of pro-apoptotic genes (CASP3, P53, BAX) and decreased expression of BCL2, respectively, was observed for all analyzed conditions with the highest differences for EP AuNPs and ELFEMF 50 mT/30 min in comparison to control cells. The highest content of cells arrested in G2/M phase was observed in ELFEMF-treated cells for 30 min both at 25 or 50 mT, while the cells treated with EP AuNPs or ELFEMF 50 mT/15 min showed highest ratios of apoptotic cells. HE staining of electroporated cells and cells exposed to ELFEMF's low and higher frequencies for different times showed nuclear pleomorphic cells. Numerous apoptotic bodies were observed in the irregular cell membrane of neoplastic and necrotic cells with mixed euchromatin and heterochromatin. Our observations indicate that treatment of Hep-2 laryngeal cancer cells with ELFEMF for 30 min at 25-50 mT and EP Au-NPs can cause cell damage inducing apoptosis and cell cycle arrest.

Size Matters: Ultra-small and Filterable Microorganisms in the Environment

Ultra-small microorganisms are ubiquitous in Earth’s environments. Ultramicrobacteria, which are defined as having a cell volume of <0.1 μm3, are often numerically dominant in aqueous environments. Cultivated representatives among these bacteria, such as members of the marine SAR11 clade (e.g., “Candidatus Pelagibacter ubique”) and freshwater Actinobacteria and Betaproteobacteria, possess highly streamlined, small genomes and unique ecophysiological traits. Many ultramicrobacteria may pass through a 0.2-μm-pore-sized filter, which is commonly used for filter sterilization in various fields and processes. Cultivation efforts focusing on filterable small microorganisms revealed that filtered fractions contained not only ultramicrocells (i.e., miniaturized cells because of external factors) and ultramicrobacteria, but also slender filamentous bacteria sometimes with pleomorphic cells, including a special reference to members of Oligoflexia, the eighth class of the phylum Proteobacteria. Furthermore, the advent of culture-independent “omics” approaches to filterable microorganisms yielded the existence of candidate phyla radiation (CPR) bacteria (also referred to as “Ca. Patescibacteria”) and ultra-small members of DPANN (an acronym of the names of the first phyla included in this superphyla) archaea. Notably, certain groups in CPR and DPANN are predicted to have minimal or few biosynthetic capacities, as reflected by their extremely small genome sizes, or possess no known function. Therefore, filtered fractions contain a greater variety and complexity of microorganisms than previously expected. This review summarizes the broad diversity of overlooked filterable agents remaining in “sterile” (<0.2-μm filtered) environmental samples.

Clinicopathologic Study of Ewingandrsquo;s Sarcoma in 82 Sudanese Patients Diagnosed at Histo Centre/ Khartoum (2013-2018)

Ewing’s sarcoma is the second most common childhood bone neoplasm, which is considered rare in African population. This is the first study in Sudan. The aim of the study is to demonstrate the clinical and histo-morphological findings of these tumours and to compare the results with similar studies. A retrospective cross-sectional study was done, conducted over 6 years’ period on 82 cases diagnosed as ES/PNET at Histo-Centre located in Khartoum state. Twenty-six of them were found to have CD99 immune-stain and ten cases have PAS special stain, data were collected, H and E, PAS and CD99 stained slides were examined by light microscope. Data analysis showed that the peak incidence of ES/PNET occurs between 5 to <18 years (58.5%) with a Mean age: 16.9 years. It shows obvious male predominance accounting for 64.6% of cases. The majority of cases are intra osseous tumour, with only 11% of cases are extra osseous, the lower limb is the most common site accounting for 54.9% of the cases, followed by the flat bones of the pelvis 13.4% with the vertebra and the skull being the least common sites accounting for 1.2% each. In clinical presentation 74.4% presented with localized swelling, 34% as localized pain, Fever and pathological fractures were the presentation in 2.4% and 3.7% respectively and none of the study population were presented as metastatic disease. Microscopic examinations of the H and E stained slides showed that 57.3% of the cases are ES devoid of neural differentiation while the rest are PNET showing some degree neural differentiation devoid of neural differentiation, among PNET cases, Homer Wright rosette were the most common rosettes in about 70% of cases. Focal areas of atypical features were found in 15.9% of cases with the pleomorphic large cells and spindle cells being the most commonly encountered features. Necrosis is present in 52.4% of cases. PAS was positive in 9 out of the 10 cases. While all the 26 cases with CD99 stained slides were positive, only 61.5% showed diffuse strong membranous positivity. In Conclusion although the demographic features and clinical presentations of ES/PNET in the study population were found to be similar to other global and regional studies finding 82 cases/6 years in single centre makes its rarity among African nations doubtful.

Pan-p63 but not ΔNp63 (p40) expression in undifferentiated carcinoma of the pancreas.

Undifferentiated carcinoma of the pancreas (UC) is a carcinoma without a definitive direction of differentiation. Tumour protein p63 is a regulator of squamous phenotype, which may also be engaged in tumour development. N-terminal isoforms of p63 are TAp63 and ΔNp63. Pan-p63 antibodies are able to detect both isoforms, whereas p40 antibodies recognise the ΔNp63 isoform only. The aim of the study was to describe pan-p63/p40 immunohistochemical expression patterns in pancreatic neoplasms: UC, ductal adenocarcinomas, neuroendocrine tumours, neuroendocrine carcinomas, serous cystic neoplasms, and solid pseudopapillary neoplasms. DAK-p63 and BC28 antibodies were used for pan-p63 and p40 detection, respectively. Moderate-to-strong pan-p63 was found in anaplastic (pleomorphic giant cell) UC (n = 4), sarcomatoid UC (n = 2), UC with osteoclast-like giant cells (n = 3), and ductal carcinomas with partial squamous differentiation. Weak and focal pan-p63 expression was found in monomorphic UC (n = 3) and in the majority of neuroendocrine carcinomas (6/7 cases). Pan-p63 expression was infrequent in ductal carcinomas without squamous differentiation and in neuroendocrine tumours. Serous cystic and solid pseudopapillary neoplasms were pan-p63-negative. Ductal carcinomas with partial squamous differentiation were the only tumours with evident p40 expression. Pan-p63(+)/p40(-) immunohistochemical status may be supportive for UC diagnosis. The pan-p63 expression was not equivalent to squamous differentiation in pancreatic neoplasia.

Primary amelanotic melanoma of the cervix: A case report with review of literature

Primary malignant melanoma arising from the uterine cervix is extremely rare. It may be misdiagnosed, especially when amelanotic, in which case immunohistochemistry is useful in reaching the diagnosis. We present a case of 67-year-old postmenopausal patient presenting with bleeding per vaginum. Per speculum examination revealed an ulceroproliferative growth involving the cervix. Microscopic examination showed sheets of pleomorphic cells with prominent eosinophilic nucleoli and intranuclear inclusion. On histopathological examination, possibilities of poorly differentiated carcinoma and amelanotic melanoma were kept. On immunohistochemical analysis, tumor cells were strongly positive for HMB-45, S-100, and vimentin. Keeping in view the histopathological and immunohistochemical findings, diagnosis of amelanotic melanoma of cervix was given. Pertinent literature is also reviewed.

Prognostic significant of osteoclast-like giant cells in undifferentiated carcinoma of pancreas

Presenter: Ryosuke Misawa MD, PhD | SUNY Downstate Medical Center Brooklyn Background: Undifferentiated carcinoma of the pancreas (UDC) is defined as containing large eosinophilic pleomorphic cells and/or ovoid-to-spindle-shaped cells and has a poor prognosis. On the other hand, undifferentiated carcinoma with osteoclast-like giant cells of the pancreas (UCOG) comprises osteoclast-like giant cells (OGCs) is suggested to have a better prognosis. However, the number of cases are limited and only case reports and their reviews are available in English literature. The aim of this study was to evaluate the prognostic significance of OGCs using the national cancer database. Methods: A retrospective analysis investigated the US National Cancer Data Base and evaluated patients who were diagnosed with UDC or UCOG from 2004 to 2014. Prognosis, demographic, and clinical features were compared between the two groups. Results: The study identified 1681 patients, 255 were in the UCOG group and 1426 were in the UDC group. The two groups were well matched in demographic profile including age ( 67.3 year-old vs 66.3 year-old, p = 0.146 ), sex ( male 56.9 % vs 57.4 %, p = 0.834 ), race distribution, and co-morbidities ( Charlson/Deyo Score ). The location of the UCOG group tumor were more common in the body and tail of the pancreas compared to the UDG group ( 32.0% vs 24.9%, p = 0.045 ), and as a consequence, more distal pancreatectomies were performed in the UCOG group ( 13.7% vs 7.2%, p = 0.011 ). Institution of chemotherapy ( 32.6% vs 29.3%, p = 0.12 ) and radiotherapy ( 20.0% vs 20.4%, p = 0.678 ) were similar in both groups. The size of the tumor at the time of diagnosis was larger in the UDC group ( 18.7 cm vs 19.7, p = 0.018 ). Lymph node metastasis were far more evident in the UDC group ( 61.7% vs 28.2%, p < 0.01 ). Kaplan-Meier analysis demonstrated superior survival for the UCOG group when compared to the UDC group ( mean OS 16.15 months vs 31.95 months, p = < 0.01 ). Conclusion: The clear prognostic significance of OGC is evident in undifferentiated carcinoma of the pancreas. Continual accumulation of UDC and UCOG cases in a database would clarify the effectiveness of surgical and neoadjuvant/adjuvant treatments in the future.

The role of immunohistochemistry for diagnosing undifferentiated malignancy in the head and neck region

Background: Undifferentiated tumours in the head and neck region are not uncommon. They can arise from different sites like in mucosa as well as in salivary glands, soft tissues or lymph nodes. Histopathological examination plays a central role in the diagnosis but difficulties arise with some tumours which are poorlydifferentiated due to their high inter- and intra-observer variability. In those cases, immunohistochemistry has greatly assisted to diagnose the tumours that cannot be accurately identified using routine histopathological procedures. The correct histopathological diagnosis is essential especially in case of malignant tumourwhere subsequent specific therapy is required. The aim of this study was to determine the role of immunohistochemistry for diagnosing undifferentiated malignancy in the head and neck region.&#x0D; Methods: This cross-sectional study was conducted during July 2014 to June 2015. A total of 35 Bangladeshi patient’s specimens of head and neck swelling were collected from two renowned laboratories in Dhaka city.These cases were diagnosed as undifferentiated tumour in histopathological examination.Standard protocol was followed for immunohistochemistry.Then primary immunohistochemical panel which included the markers for Epithelial CK (AE1/AE3),mesenchymal marker (Vimentin) and lymphoid marker (LCA) were used.The cases which were not resolved by primary immunohistochemistry panel, the second panel was applied for further sub classification (Desmin, Chromogranin, CK20, CEA, CD20, CD30, HMB45, NSE). Based on interpretation of immunohistochemical findings final diagnoses were made.Data analysis was performed using the Statistical Package for the Social Sciences for Windows version 22.0 (SPSS, Chicago, Illinois, USA).&#x0D; Results: A total of 35 undifferentiated tumorsof head-neck region were studied.The mean age was 46.3±17.6 years and male to female ratio was 4.8:1. The majority 13 (37.1%) patient had cervical lymphadenopathy, 11 (31.4%) had neck mass and 4(11.4%) had in nasal/sinonasal mass.Regarding histopathologicalcell types, round cell was 21 (60.0%), spindle cell 6 (17.1%), pleomorphic cells 6(17.1%) and epithelioid cells 2(5.7%). In initial histopathological examination, majority 25(71.4%) were undifferentiated malignant neoplasm, 8(22.8%) were metastatic undifferentiated carcinoma, 1(2.9%) was pleomorphic sarcoma and the remaining 1(2.9%) had malignant adnexal tumour. By application of immunohistochemistry, most (33, 94.3%) of the cases were resolved and the two cases remained unresolved. Among the resolved cases majority (15, 45.5%) were lymphoma, 4(12.1%) were metastatic carcinoma, 3(9.1%) were Ewing’s sarcoma and malignant melanoma were found in 3(9.1%) cases.&#x0D; Conclusion: This study supports that the immunohistochemical technique has a fundamental role in the investigation of undifferentiated tumour origin, to determine the correct guidance for treatment and improving the prognosis for head and neck tumour patients.&#x0D; Birdem Med J 2020; 10(1): 48-53

Exquisite Tumor Targeting by Salmonella A1-R in Combination with Caffeine and Valproic Acid Regresses an Adult Pleomorphic Rhabdomyosarcoma Patient-Derived Orthotopic Xenograft Mouse Model

Adult pleomorphic rhabdomyosarcoma (RMS) is a rare and malignant mesenchymal tumor. Recently, we developed a patient-derived orthotopic xenograft (PDOX) model of adult pleomorphic RMS. In the present study, we evaluated the efficacy of tumor-targeting Salmonella typhimurium (S. typhimurium) A1-R combined with caffeine (CAF) and valproic acid (VPA) on the adult RMS PDOX. An adult pleomorphic RMS cell line was established from the PDOX model. Cell survival after exposure to CAF and VPA was assessed, and the IC50 value was calculated for each drug. The RMS PDOX models were randomized into five groups: untreated control; tumor treated with cyclophosphamide (CPA); tumor treated with CAF + VPA; tumor treated with S. typhimurium A1-R; and tumor treated with S. typhimurium A1-R + CAF + VPA. Tumor size and body weight was measured twice a week. VPA caused a concentration-dependent cytocidal effect. A synergistic effect of combination treatment with CAF was observed against the RMS cell line. For the in vivo study, all treatments significantly inhibited tumor growth compared with the untreated control. S. typhimurium A1-R combined with VPA and CAF was significantly more effective than CPA, VPA combined with CAF, or S. typhimurium A1-R alone and significantly regressed the tumor volume compared with day 0. These results suggest that S. typhimurium A1-R together with VPA and CAF could regresses an adult pleomorphic RMS in a PDOX model and therefore has important future clinical potential.

ORAL AND MAXILLOFACIAL OSTEOSARCOMA: CASE REPORT

A 43-year-old female patient attended the oral maxillofacial and traumatology service presenting volume increase in the right hemiface with painful symptomatology. At the intraoral examination, an exophytic lesion involving the alveolar ridge and palate region was observed, extending to the right nostril. The lesion showed a partially ulcerated surface, with purulent secretion, firm consistency, and normal mucosal color. Computed tomography revealed a mixed appearance of hyperdensity and ill-defined hypodensity in the maxillary region with extension to the zygoma and the right orbit. Because of poor oral hygiene and a history of regression with antibiotic therapy, a clinical diagnosis of actinomycosis was issued, and incisional biopsy of the lesion was subsequently performed. In the microscopic analysis, proliferation of pleomorphic and atypical malignant mesenchymal cells was associated with deposition of osteoid matrix, thus confirming the histopathologic diagnosis of osteosarcoma. The patient was referred to the State Reference Center for appropriate treatment.