Round Cell Research Articles

Features of fat body development in the body of honey bees

The fat body is considered an organ that contains a conglomerate of proteins and fats and acts similar to the liver of mammals. It is loose tissue that is involved in some metabolic functions. One of these functions is to store and release energy in response to energy needs. The article presents data on the development of trophocytes of the fat body of the honey bee and factors that affect the process of lipolysis and lipogenesis. These processes are related to the technological methods of keeping bee colonies. The purpose of the work was to study patterns and factors that affect the growth and development of the structural elements of the fat body. For this, honeycombs with sealed brood were taken from the nest for incubation. After leaving the cells, the selected bees were placed in the nest of the bee colony. Marked bees were selected every 3 days. Histological preparations of various parts of the fat body were studied up to 21 days inclusive. According to research, trophocytes differ from enocytes in their larger size. These rounded cells have a distinct membrane and granular cytoplasm. In the cytoplasm, mostly oval-shaped vacuoles were found, which are filled with energetic and plastic substances, while others are empty. The age-related dynamics of the length of fat body trophocytes was studied. It was found that at the exit from the cell, they are the smallest and amount to an average of 50.12 ± 3.18 μm. With an increase in the activity of enzymes of the intestinal tract, an increase in the morphometric indicators of fat cells is observed. The maximum indicators of the length of trophocytes of the pericardial zone of the fat body were found in 15 daily bees. Measurement of the length of fat cells during this period shows that compared to diurnal bees, trophocytes are larger by 34.6 % (Р < 0.001). After 14 days, a gradual regression of the morphometric indicators of trophocytes is noted. The article analyzes the role of the fat body in the life of the honey bee.

Machine Learning–Supported Diagnosis of Small Blue Round Cell Sarcomas Using Targeted RNA Sequencing

Small blue round cell sarcomas (SBRCSs) are a heterogeneous group of tumors with overlapping morphologic features but markedly varying prognosis. They are characterized by distinct chromosomal alterations, particularly rearrangements leading to gene fusions, whose detection currently represents the most reliable diagnostic marker. Ewing sarcomas (ESs) are the most common SBRCSs, defined by gene fusions involving EWSR1 and transcription factors of the ETS gene family, while the most frequent non-EWSR1-rearranged SBRCSs harbor a CIC rearrangement. Unfortunately, the identification of CIC::DUX4 translocation events, the most common CIC rearrangement, is challenging with current methods. Here, a machine-learning approach to support SBRCS diagnosis relying on gene expression profiles measured via targeted sequencing is presented. The analyses on a curated cohort of 69 soft tissue tumors (STT) showed markedly distinct expression patterns for SBRCS subgroups. A random forest (RF) classifier trained on ES and CIC-rearranged cases predicted probabilities of being CIC-rearranged > 0.9 for CIC-rearranged-like sarcomas and < 0.6 for other SBRCS. Testing on a retrospective cohort of 1335 routine diagnostic cases identified 15 candidate CIC-rearranged tumors with a probability > 0.75, all of which supported by expert histopathological reassessment. Furthermore, the multi-gene RF classifier appeared advantageous over using high ETV4 expression alone, previously proposed as surrogate to identify CIC rearrangement. Taken together, the expression-based classifier can offer valuable support for SBRCS pathological diagnosis.

Next-generation sequencing for pediatric CNS tumors: does it add value in a middle-income country setup?

Advances in molecular diagnostics led to improved targeted interventions in the treatment of pediatric CNS tumors. However, the capacity to test for these is limited in LMICs, and thus their value needs exploration. We reviewed our experience with NGS testing (TruSight RNA Pan-Cancer-seq panel) for pediatric CNS tumors at KHCC/Jordan (March/2022-April/2023). Paraffin blocks' scrolls were shipped to the SickKids laboratory based on the multidisciplinary clinic (MDC) recommendations. We reviewed the patients' characteristics, the tumor types, and the NGS results' impact on treatment. Of 237 patients discussed during the MDC meetings, 32 patients (14%) were included. They were 16 boys and 16 girls; the median age at time of testing was 9.5 years (range, 0.9-21.9 years). There were 21 samples sent at diagnosis and 11 upon tumor progression. The main diagnoses were low-grade-glioma (15), high-grade-glioma (10), and other histologies (7). Reasons to request NGS included searching for a targetable alteration (20) and to better characterize the tumor behavior (12). The median turnaround time from samples' shipment to receiving the results was 23.5 days (range, 15-49 days) with a median laboratory processing time of 16 days (range, 8-39 days) at a cost of US$1,000/sample. There were 19 (59%) tumors that had targetable alterations (FGFR/MAPK pathway inhibitors (14), checkpoint inhibitors (2), NTRK inhibitors (2), and one with PI3K inhibitor or IDH1 inhibitor). Two rare BRAF mutations were identified (BRAFp.G469A, BRAFp.K601E). One tumor diagnosed initially as undifferentiated round cell sarcoma harbored NAB2::STAT6 fusion and was reclassified as an aggressive metastatic solitary fibrous tumor. Another tumor initially diagnosed as grade 2 astroblastoma grade 2 was reclassified as low-grade-glioma in the absence of MN1 alteration. NGS failed to help characterize a tumor that was diagnosed histologically as small round blue cell tumor. Nine patients received targeted therapy; dabrafenib/trametinib (6), pembrolizumab (2), and entrectinib (1), mostly upon tumor progression (7). In this highly selective cohort, a high percentage of targetable mutations was identified facilitating targeted therapies. Outsourcing of NGS testing was feasible; however, criteria for case selection are needed. In addition, local capacity-building in conducting the test, interpretation of the results, and access to "new drugs" continue to be a challenge in LMICs.

Primary diffuse large B-cell lymphoma of the prostate: Histopathological diagnosis of a rare entity in the prostate.

Lymphoma of the prostate is rare whether it is primary extranodal lymphoma or secondary involvement of the prostate by primary lymphoma elsewhere. Of all the lymphomas of the prostate, primary lymphomas of the prostrate are very rare. Although less frequent, it should be a differential diagnosis when evaluating prostate tumors. Here, we report a case of a 61-year-old man who presented with hematuria with clot retention. A cystoscopy with clot removal and transurethral resection of the prostate (TURP) was performed. This biopsy was sent for histopathological examination at an external center, where a diagnosis of benign prostatic hyperplasia was given. Fifteen days later, the patient presented with hematuria again. On examination, clots were present in the bladder. There was significant prostatomegaly. A re-resection of the prostate was performed and sent for another review to us. The biopsy was reported as high-grade round cell neoplasm, most likely lymphoma. Immunohistochemistry (IHC) was recommended for confirmation. Tumor cells showed immunoreactivity for CD20, B-cell Lymphoma (BCL)-2, BCL-6, Myelocytomatosis (c-Myc), and multiple myeloma 1 (MUM1). Cluster Differentiation (CD)10 was negative. Kiel-67 was high. A final diagnosis of double-expressor diffuse large B-cell lymphoma (DLBCL) of non-germinal center type was made. We share this case to emphasize the fact that primary lymphoma of the prostate is primarily a histopathological diagnosis as the clinical presentation is not unique. Owing to its rarity, the clinical and histopathological suspicion is low. Hence, keeping the differential in mind while evaluating prostate biopsy is beneficial in a timely diagnosis of the entity as the management of prostatic carcinoma and lymphoma is different.

Extragenital transmissible venereal tumor in dog: case report

Transmissible venereal tumor (TVT) is a round cell neoplasm that affects dogs, mainly with access to the street, because tumor implantation happens during coitus or by intimate social interaction with another sick canid. It mainly affects the genital region, but there are extragenital reports. Thus, this work aims to report a case of TVT in the eyelid and cervical region of a dog, male, non-castrated, adult, SRD, with access to the street, without previous history, positive for cynomosis, which presented ectoparasites, penile bloody secretion with bulb volume increase, nodular tumor ulcerated in cervical region, left eye with third eyelid volume increase and some small nodules in upper eyelid. Through cytology and histopathology, TVT was diagnosed in the samples analyzed. For treatment, five sessions of chemotherapy with Vincristine sulfate were carried out, which the animal responded with an improvement in clinical signs one week after the first session. Therefore, the importance of cytology and histopathology for diagnostic confirmation of TVT and of care in the analysis of ocular and skin tumors is emphasized, because, even if the occurrence of this neoplasia is common only in the genital region, it can present itself in other locations.

Tongue plasmacytoma in a dog treated with adjuvant metronomic melphalan

A 9-year-old female crossbred dog was referred due to a tongue neoformation. Marginal resection was performed to proceed with histopathological examination that indicated a malignant round cell neoplasia, and immunohistochemical staining diagnosed the mass as plasmacytoma. The owner refused another surgical intervention to extend margins and approach another mass that was verified on the ventral surface of the base of the tongue. Thus, the patient underwent three months of metronomic chemotherapy with melphalan. The disease progression was not detected at two years of follow-up.

Multicentric lymphoma in a donkey with intestinal and bone marrow involvement

BackgroundLymphoma is a common neoplasm in horses but is reported much less commonly in donkeys. In this case report, we describe the macroscopic, microscopic and immunohistochemical features of a multicentric lymphoma with intestinal and bone marrow involvement.Case presentationA geriatric female donkey with history of chronic lameness was found dead. Post-mortem examination revealed advanced emaciation, periodontal disease, left front foot laminitis and multiple, soft, white to yellow tan intestinal transmural masses, up to 12 cm in diameter. Cytology suggested a round cell intestinal neoplasm. The femur of the left hint limb was double the size of the normal contralateral, with suspected neoplastic infiltration and replacement of bone marrow and bone. Histologically we diagnosed a lymphoma in the intestine and left femur. Immunohistochemically, the neoplastic cells showed CD3 immunolabelling, supporting a diagnosis of a multicentric T-cell lymphoma.ConclusionsTo the authors’ knowledge, this is the first time multicentric lymphoma is diagnosed in donkeys. Further studies of the genetic background, clinical, laboratory, histopathologic, and immunohistochemical, as well as the pathogenesis of lymphoma, is needed to better understand the uniquely low frequency of this neoplasm in donkeys.

Terato-Neuroendocrine Tumour of Testis: Review and Update

Neuro-endocrine tumours commonly emanate from intestinal and respiratory epithelium. Neuro-endocrine tumours could also on rare occasions afflict various organs of the human body. Neuro-endocrine tumour afflicting the testis is an uncommon tumour which does account for far less than one percent (&lt;1%) of all tumours of testis. To the knowledge of the author, less than 100 cases of primary neuro-endocrine tumour of the testis had been reported so far in the global literature. The reported cases of neuro-endocrine tumour of testis have ranged between 10 years and 83 years of age and the reported incidence rate had been noted to be higher within the fifth and sixth decade of life. Majority of patients who are afflicted by neuro-endocrine tumour of testis do tend to manifest with unilateral painless testicular mass. Sixteen percent (16%) of patients who are afflicted by neuro-endocrine tumour of testis do manifest with symptoms of neuroendocrine tumour syndrome. Eleven percent (11%) of primary neuroendocrine tumours of testis tend to be diagnosed initially with a contemporaneously-associated metastasis. Neuro-endocrine tumours of the testis occur as a primary testicular neuroendocrine tumour, or they may be metastatic neuroendocrine tumour to the testis with the primary tumour originating from elsewhere in the body especially from the gastrointestinal or cardiorespiratory tract system. Primary neuro-endocrine tumour of testis, could be further sub-divided into: (a) primary pure neuroendocrine tumour of testis; and (b) neuro-endocrine tumour of testis contemporaneously associated with teratoma or dermoid/epidermoid cysts. Once a neuro-endocrine tumour of testis is diagnosed, it is pivotal that clinicians should exclude a metastatic neuroendocrine tumour to the testis with the primary tumour originating from elsewhere within the human body. It has been iterated that the presence of teratomas elements within the neuro-endocrine tumour of testis does sufficiently rule out the primary site outside the testis. In order to accurately diagnose the neuro-endocrine tumour of the testis, it has been advised that it is pertinent to submit the tumour mass entirely and to look for any additional lineage of differentiation. Twenty five percent of primary neuro-endocrine tumours of testis are associated with teratoma. Histogenesis of pure neuroendocrine tumour of the testis is yet to be clarified. Two postulates had been propounded in relation to the mode origin of neuroendocrine tumour of testis including: (1) Emanation from a teratoma which is the germ cell origin postulate; (2) Emanating from argentaffin cells that are located within crypts of Lieberkühn .A study related to three pure testicular neuroendocrine tumours of testis had indicated that pure neuroendocrine tumour of testis might have different genetic background other than germ cell tumour.[11] Histopathology examination of specimens of neuro-endocrine tumour of testis has tended to demonstrate tumour cells that tend to be typified by the presence of nests of small round cells with uniform nuclei forming small acini and rosettes or sheets. The cells tend to contain eosinophilic granules within the cytoplasm and granular chromatin within the nuclei. The tumour cells do exhibit positive staining for neuroendocrine markers like chromogranin and synaptophysin as illustrated in detail in the article. Radical orchidectomy is the most adopted option of treatment; nevertheless, necessitation for the undertaking of provision of adjuvant treatment depending upon histological grading has not been clarified. Because of the possibility of development of local recurrence and or distant metastases after a long time, it is pivotal for all clinicians to carefully follow-up their patients over a long-period of time with clinical, laboratory testing and radiology-imaging testing follow-ups in order to establish early diagnosis of any local recurrence or distant metastasis to be able provide further treatment of curative intent early. There is also the possibility that neuro-endocrine tumours of testis had so far been under-reported due to mis diagnosis of the tumour upon pathology examination of the testicular tumour or the correct set of monoclonal and polyclonal antibodies had not been utilized in the immunohistochemistry study assessment of the tumours. It would be advised that all clinicians and pathologists globally should have a high index of suspicion for a neuro-endocrine tumour of testis in order to establish prompt diagnosis of the tumour and to provide appropriate assessment, treatment and follow-up of all their patients.

Chromoplexy Is a Frequent Early Clonal Event in EWSR1-Rearranged Round Cell Sarcomas That Can Be Detected Using Clinically Validated Targeted Sequencing Panels.

Chromoplexy is detectable using targeted NGS in a substantial portion of EWSR1-rearranged round cell sarcomas as an early and persistent clonal event, expanding the genomic complexity of fusion-associated sarcomas.

Small cell osteosarcoma versus fusion-driven round cell sarcomas of bone: retrospective clinical, radiological, pathological, and (epi)genetic comparison with clinical implications.

Small cell osteosarcoma (SCOS), a variant of conventional high-grade osteosarcoma (COS), may mimic fusion-driven round cell sarcomas (FDRCS) by overlapping clinico-radiological and histomorphological/immunohistochemical characteristics, hampering accurate diagnosis and consequently proper therapy. We retrospectively analyzed decalcified formalin-fixed paraffin-embedded (FFPE) samples of 18 bone tumors primarily diagnosed as SCOS by methylation profiling, fusion gene analysis, and immunohistochemistry.In eight cases, the diagnosis of SCOS was maintained, and in 10 cases it was changed into FDRCS, including three Ewing sarcomas (EWSR1::FLI1 in two cases and no identified fusion gene in the third case), two sarcomas with BCOR alterations (KMT2D::BCOR, CCNB3::BCOR, respectively), three mesenchymal chondrosarcomas (HEY1::NCOA2 in two cases and one case with insufficient RNA quality), and two sclerosing epithelioid fibrosarcomas (FUS::CREBL3 and EWSR1 rearrangement, respectively).Histologically, SCOS usually possessed more pleomorphic cells in contrast to the FDRCS showing mainly monomorphic cellular features. However, osteoid was seen in the latter tumors as well, often associated with slight pleomorphism. Also, the immunohistochemical profile (CD99, SATB2, and BCOR) overlapped.Clinically and radiologically, similarities between SCOS and FDRCS were observed, with by imaging only minimal presence or lack of (mineralized) osteoid in most of the SCOSs.In conclusion, discrimination of SCOS, epigenetically related to COS, versus FDRCS of bone can be challenging but is important due to different biology and therefore therapeutic strategies. Methylation profiling is a reliable and robust diagnostic test especially on decalcified FFPE material. Subsequent fusion gene analysis and/or use of specific immunohistochemical surrogate markers can be used to substantiate the diagnosis.

Isofagomine Inhibits Multiple TcdB Variants and Protects Mice from Clostridioides difficile-Induced Mortality.

Clostridioides difficile causes life-threatening diarrhea and is one of the leading causes of nosocomial infections. During infection, C. difficile releases two gut-damaging toxins, TcdA and TcdB, which are the primary determinants of disease pathogenesis and are important therapeutic targets. Once in the cytosol of mammalian cells, TcdA and TcdB use UDP-glucose to glucosylate host Rho GTPases, which leads to cytoskeletal changes that result in a loss of intestinal integrity. Isofagomine inhibits TcdA and TcdB as a mimic of the glucocation transition state of the glucosyltransferase reaction. However, sequence variants of TcdA and TcdB across the clades of infective C. difficile continue to be identified, and therefore, evaluation of isofagomine inhibition against multiple toxin variants is required. Here, we show that isofagomine inhibits the glucosyltransferase domain of multiple TcdB variants and protects TcdB-induced cell rounding of the most common full-length toxin variants. Furthermore, we demonstrate that isofagomine protects against C. difficile-induced mortality in two murine models of C. difficile infection. Isofagomine treatment of mouse C. difficile infection also permitted the recovery of the gastrointestinal microbiota, an important barrier to preventing recurring C. difficile infection. The broad specificity of isofagomine supports its potential as a prophylactic to protect against C. difficile-induced morbidity and mortality.

Acute Lymphoblastic Leukemia (ALL) as a secondary malignancy in adolescent patient after completion of Ewing's sarcoma chemotherapy: a case report

OBJECTIVE: To report an adolescent patient who was diagnosed with Acute lymphoblastic leukemia (ALL) after completing Ewing sarcoma (ES) therapy. CASE: A 14 years old girl complained of left crusis tumor since 4 months before hospitalized. Tumor biopsy showed a round cell tumor, as well as immunohistochemistry which showed a positive CD99, consistent with ES. The patient had received 14 cycles chemotherapy for 7 months with regimens vincristine 1.5 mg/m2, cyclophosfamide 1200 mg/m2, dactinomycin 0.5 mg/m2, and doxorubicin 30 mg/m2. Between chemotherapy, patient also was given granulocyte colony-stimulating factor several times. Pasien also undergo 31 times radiation therapy with total 31x1.8 Gy (55.8 Gy). Two months after last chemotherapy for Ewing sarcoma, she was admitted again with prolonged fever, and pallor.Laboratorium showed leukocytosis (88.86x103uL), monocytosis (41%), severe neutropenia (0.39x103uL), severe normochromic normocytic anemia (4,3 g/dL), severe thrombocytopenia (4x103uL), reticulocytopenia (0,46%), with lymphoblast in peripheral blood smear. Bone marrow aspiration result showed lymphoblast infiltration 60% with heterogen morphology, no cluster cell non hematopoetic in bone marrow, consisten with acute lymphoblastic leukemia (ALL-L2) suspected secondary malignancy from Ewing’s Sarcoma. Patient underwent chemotherapy for high risk ALL at induction phase with combination of vincristin, daunorubicin, L-asparginase, metotrexate high dose, prednison, and 6-mercaptopurine. After completing induction phase, the patient’s condition get worsening and later died. CONCLUSION: ES survivors are at significant risk for secondary malignancies. Leukimia is one of which the most common secondary malignancies. Young children are particularly susceptible to the mutagenic effects of both chemotherapy and radiotherapy, as proven by several studies.

Clinicopathological Features of Three Rare EWSR1::NFATC2 Sarcomas of Bone and Soft Tissues.

Certain undifferentiated round cell sarcomas displaying EWSR1::NFATC2 fusion have recently been reported, mostly in the bones. This report presents clinicopathological features of 3 additional EWSR1::NFATC2 fusion sarcomas of bone and soft tissues. We present 2 soft tissue and 1 bone tumors: A 62-year-old man with pain and a slowly growing, 8-cm-sized soft tissue mass in the anterolateral compartment of his right calf, along with multiple pulmonary metastatic lesions; a 63-year-old man with a 5-cm sized axillary mass of 4 months duration and a cystic renal mass; and a 53-year-old man with a complaint of leg pain was found to have a 2-cm diameter, intramedullary, lytic mass in the diaphysis of his left femur. Microscopic examination of the tumors in all patients revealed round to epithelioid cells arranged in cords and trabeculae in a myxohyaline stroma. Immunohistochemically, the tumor cells were positive for MIC2/CD99 (3/3), EMA (3/3), NKX3.1 (3/3), NKX2.2 (2/2), CD10 (2/2), and aggrecan (1/1), while negative for S100P and GFAP. Various keratins were also negative except focal AE1/AE3 positivity in the third tumor. By fluorescence in-situ hybridization, 2 tumors (#1 and #3) revealed EWSR1 gene rearrangement and amplification. Furthermore, 2 tumors (#1 and #2) displayed EWSR1ex8::NFATC2ex3 fusion with next-generation sequencing (NGS). The first patient was offered chemotherapy. However, he died of pulmonary metastasis. This report highlights the value of combining histopathological features and immunostains such as NXK3.1, NKX2.2, CD10, and aggrecan, along with EWSR1 testing for triaging these tumors for rare gene fusions by NGS that has prognostic implications.

(302) Intertest Reliability of Seminal Round Cells and Leukocytes Without Treatment in Men Presenting for Infertility

Abstract Introduction Pyospermia may be due to infection or inflammation along the male reproductive tract. Semen round cells and leukocytes can have a major impact on the management of infertility yet we have a poor understanding of the natural history of patients with seminal round cells or pyospermia. Objective We aimed to assess the intertest reliability of seminal leukocytes and round cells within serial semen analyses in patients without treatment. Methods We conducted a retrospective review of men presenting to a high-volume fertility center for evaluation who had multiple semen analyses assessed for leukocytes between 2/1/2022 and 1/1/2023. Men with azoospermia or who received treatment for pyospermia including antibiotics or surgery in the interval between the tests were excluded. We recorded round cells, the presence of leukocytes, and days between tests. The presence of leukocytes was detected using QwikCheck™ Test Strips which report a positive result if the concentration of leukocytes was greater or equal to 1 million/ml. A Bland-Altman assessment of agreement was conducted to assess the agreement of the number of round cells between the first and second semen analysis. The limits of agreement (LoA) represent the 95% confidence intervals between which we expect most of the differences between measurements. The number of round cells was log-transformed prior to analysis as they were positively skewed. A Cohen Kappa test was used to assess the agreement between the presence of leukocytes in the first sample and the second sample. Results One hundred patients were included in the analysis. The mean time between tests was 75 days (SD: 68.3 days) ranging from 1 to 285 days. There was a median of 2.95 million round cells in the initial semen analysis and 2.45 million round cells in the follow-up semen analysis. Nineteen patients had positive leukocytes on the initial sample and fifteen on the second sample. Eight patients had persistently positive leukocytes. The assessment of agreement for the round cells in the initial and follow-up semen analysis shows a lower LoA of 22% and an upper LoA of 582% meaning that most measurements of round cells in the follow-up tests are estimated to be between 22% and 582% of the initial measurement. The wide range of LoAs suggests poor agreement between tests. The kappa for the presence of leukocytes in the first and second sample was 0.36 (CI 0.13-0.60) which would be considered a fair agreement. Conclusions Round cells and semen leukocytes are poorly understood but are used to aid in the diagnosis and management of male infertility. The number of round cells and the presence of pyospermia between samples varies greatly in patients even without treatment. Providers should consider these discrepancies prior to initiating treatment of pyospermia in men presenting for infertility. Further study is needed to gain a better understanding of how round cells and semen leukocytes should influence management of men with infertility. Providers should be cautious when using the presence of round cells or pyospermia to influence treatment as there is poor consistency between tests even without pyospermia treatment. Disclosure No.

(378) Shorter Abstinence is Associated with a Decrease in Semen Round Cells but Not in Semen Leukocytes

Abstract Introduction The connection between abstinence times and semen round cells and pyospermia is not well understood. However, frequent ejaculation is a commonly prescribed treatment for pyospermia. Objective We sought to identify if a correlation exists between abstinence periods and round cells and semen leukocytes. Methods We conducted a retrospective review of men presenting to a high-volume fertility center for evaluation who had semen samples assessed for leukocytes between 2/1/2022 and 1/1/2023. Azoospermic men and men with abstinence times over 14 days were excluded. We recorded round cells, the presence of leukocytes, and days of abstinence prior to the sample. The presence of leukocytes was detected using test strips which report a positive result if the concentration of leukocytes was greater or equal to 1 million/ml. The study received IRB approval. Pearson correlations (assumes linear relationship) were used as well as generalized additive models (GAMs), which allow for a non-linear relationship between abstinence and the outcome. For analysis of semen leukocytes, a positive result was coded as 1 and a negative result was coded as 0. Round cells were log-transformed prior to calculating correlation. The estimated relationships from GAMs were plotted graphically and summarized via average marginal slopes. Results A total of 382 semen analyses were included, with a mean abstinence period of 3.67 days (SD: 2.11). The number of round cells ranged from 0.2 to 44.0 million (mean: 4.21 million, SD: 4.68), with 19.1% (n = 73) of samples testing positive for leukocytes. Days of abstinence and round cells are significantly correlated according to a Pearson correlation (r = 0.14, p = 0.006). According to the GAM, with each day increase in abstinence, a 0.52 million increase in round cells is expected (CI 0.25-0.79 million; p&amp;lt;0.05). There is an increase in round cells from 0 to 5 days of abstinence before plateauing. Beyond 10 days there is a large degree of uncertainty, due to a low number of observations. When patients with negative pyospermia tests were excluded, there is no linear correlation (r = 0.11, p = 0.36), however, the GAM appears similar to the original sample (1.2; 0.2, 2.1; p&amp;lt;0.05). Semen leukocytes and abstinence are not significantly correlated according to the Pearson correlation (r = -0.04, p = 0.38). There is little change in the probability of leukocytes being present with respect to abstinence in the GAM (-0.001; CI 2.3-2.1; p = 0.926). There is a similar increase in the degree of uncertainty over 10 days. Conclusions These results demonstrate a statistically significant, yet very minimal linear correlation between round cells and days of abstinence. There is an increasing number of round cells up to 5 days of abstinence before plateauing. However, there was no significant correlation between abstinence time and the presence of leukocytes in the semen in either model. Shorter abstinence times may have a role in reducing semen round cells, but it is unlikely to reduce pyospermia. Providers should be hesitant when suggesting an increased frequency of ejaculation to treat pyospermia. Disclosure No.

The role of cell-matrix adhesion and cell migration in breast tumor growth and progression.

During breast cancer progression, there is typically increased collagen deposition resulting in elevated extracellular matrix rigidity. This results in changes to cell-matrix adhesion and cell migration, impacting processes such as the epithelial-mesenchymal transition (EMT) and metastasis. We aim to investigate the roles of cell-matrix adhesion and cell migration on breast tumor growth and progression by studying the impacts of different types of extracellular matrices and their rigidities. We embedded MCF7 spheroids within three-dimensional (3D) collagen matrices and agarose matrices. MCF7 cells adhere to collagen but not agarose. Contrasting the results between these two matrices allows us to infer the role of cell-matrix adhesion. We found that MCF7 spheroids exhibited the fastest growth rate when embedded in a collagen matrix with a rigidity of 5.1kPa (0.5mg/mL collagen), whereas, for the agarose matrix, the rigidity for the fastest growth rate is 15kPa (1.0% agarose) instead. This discrepancy is attributable to the presence of cell adhesion molecules in the collagen matrix, which initiates collagen matrix remodeling and facilitates cell migration from the tumor through the EMT. As breast tumors do not adhere to agarose matrices, it is suitable to simulate the cell-cell interactions during the early stage of breast tumor growth. We conducted further analysis to characterize the stresses exerted by the expanding spheroid on the agarose matrix. We identified two distinct MCF7 cell populations, namely, those that are non-dividing and those that are dividing, which exerted low and high expansion stresses on the agarose matrix, respectively. We confirmed this using Western blot which showed the upregulation of proliferating cell nuclear antigen, a proliferation marker, in spheroids grown in the 1.0% agarose (≈13kPa). By treating the embedded MCF7 spheroids with an inhibitor or activator of myosin contractility, we showed that the optimum spheroids' growth can be increased or decreased, respectively. This finding suggests that tumor growth in the early stage, where cell-cell interaction is more prominent, is determined by actomyosin tension, which alters cell rounding pressure during cell division. However, when breast tumors begin generating collagen into the surrounding matrix, collagen remodeling triggers EMT to promote cell migration and invasion, ultimately leading to metastasis.

Evaluation of linear dermatoses by reflectance confocal microscopy in children.

Lichen striatus (LS), linear psoriasis (LPs), linear cutaneous lupus erythematosus (LCLE) and linear lichen planus (LLP) often have similar clinical manifestations, which makes clinical diagnosis with the naked eye difficult; therefore, they are easily misdiagnosed. The purpose of this study was to determine whether reflectance confocal microscopy (RCM) is helpful in differentiating between these four linear dermatoses in children. This retrospective study included 14 patients with LS, nine with LPs, eight with LCLE and 12 with LLP. All patients were analysed using RCM, and biopsies were collected from lesions previously imaged by RCM. For LS, the dermal papillary rings were partially absent, but when present, manifested with small, homogeneously round, bright cells and occasionally highly refractive plump cellular structures, aggregated in clusters. LPs exhibited dark cyst-like structures with small, bright, round cells aggregated at the epidermal level; at the dermal-epidermal junction, homogeneously distributed, enlarged, faint dermal papillary rings and numerous enlarged low-refractive canalicular structures were observed in the superficial dermis. LCLE and LLP exhibited similar manifestations, including epidermal disarray, almost total absence of dermal papillary rings, and various sized refractive structures densely distributed in the dermis. The key distinguishing features of LCLE were the different sized structures mainly clustered around hair follicles, while LLP demonstrated dense structures with a scattered distribution. RCM may be used to distinguish between the key features of LS, LPs, LCLE and LLP in children.

Retroperitoneal Castleman’s disease in a young Nepalese girl: A rare cause of childhood abdominal mass

Introduction: Castleman’s disease (CD), or benign angio-follicular lymph node hyperplasia, is an uncommon condition in childhood. When a child presents with a huge retroperitoneal mass and inconclusive findings on clinico-radiological evaluation or tissue sampling, management becomes exceedingly difficult. CD herein becomes an uncommon presentation of an uncommon diagnosis. Case presentation: A six-year-old girl with no past medical problems presented to the office with a slowly progressive, painless mass over the right lumbar region for a year. Abdominal ultrasound showed a well-defined oval mass in the right periumbilical region, further evaluation of which with a computed tomography scan suggested lymphoma. A preoperative core-cut biopsy could not confirm the findings and suggested a neoplastic lesion, probably an inflammatory myofibroblastic tumour or small round cell tumour. She underwent an exploratory laparotomy with in-toto excision of the mass. Intraoperatively, a solid retroperitoneal tumour measuring 8×8×6 cm was found. Histopathology and immunohistochemistry confirmed a unicentric CD of the hyaline-vascular type. At two years of follow-up, she remained asymptomatic and disease-free. Conclusion: While CD in children is rare, retroperitoneal localization of the same can further add to the diagnostic conundrum. However, if carefully considered, an en-bloc surgical resection offers complete treatment.

Extraskeletal Ewing sarcoma arising from stomach and ovary: A case series of two rare sites

Ewing sarcoma is the second most common tumor in children and young adults. Primitive neuroectodermal tumors/Ewing sarcoma family consist of small round cell of neuroectodermal origin. Ewing sarcoma commonly arises from long bones and pelvic region but extraskeletal Ewing sarcoma (EES) is a very rare tumor with highly malignant potential associated with poor outcome. We present a case series of two cases, one is from the ovary in a 24-year-old young, married, multiparous woman who presented with metastatic disease and was treated with palliative chemotherapy. The second is an Ewing sarcoma arising from the stomach in a 33-year-old male who underwent surgery followed by adjuvant chemotherapy. EES is usually an uncommon tumor and many times, the diagnosis becomes delayed and presents with metastatic disease. Immunohistochemical and molecular analysis is a must to diagnose the EES. There are no standard treatment guidelines. Surgery, various chemotherapy regimens, and radiotherapy are given for treatment. Early diagnosis and interventions are required for better outcomes.

Confocal Assessment of Pigmented-Mucosal Lesions: A Monocentric, Retrospective Evaluation of Lip and Genital Area.

Pigmentation of lip and/or genitalia is mainly due to the development of benign melanotic macules, with a less occurrence of melanocytic and other non-melanocytic lesions. Mucosal melanoma has worse prognosis compared with cutaneous counterpart, hence identification of atypical features for an early diagnosis is crucial. The aim of this study was to report further data of confocal features characterizing pigmented mucosal lesions of genital area and of the lips and test the diagnostic role of the reflectance confocal microscopy (RCM)lip score. Clinical, dermoscopic and RCM images of histologically proven pigmented lesions, involving the genital area (vulva or glans penis) and lip, were retrospectively reviewed. RCM images were evaluated for malignant criteria, and statistical analysis was conducted for categorical variables. Seventy pigmented lesions were included in the study and divided into two groups based on the body area location: lip (17) and genital area (53). Architectural disarray (P = 0.002), dendritic (P = 0.031) and roundish cells in epidermis (P < 0.0001), interpapillary dendritic cells (P = 0.039) and junctional atypical cells (P = 0.002) were associated to genital melanoma. Melanoma involving the lip was characterized by roundish cells in epidermis, a criterion found in one labial benign lesion, only (P = 0.005). Main limitations of the study are the inclusion of low melanomas and the presence of epidermal dendritic cells in melanosis and melanoma, as a confusing factor in imaging. Dermatologists should consider confocal microscopy as an adjunctive tool to dermoscopy in the differential diagnosis of pigmented mucosal lesions, especially in presence of clinical and dermoscopic findings suspicious for malignancy.