Immunoexpression of CD34, CD117, and p53 in Hypocellular Bone Marrow Disorders.

Objectives Hypocellular bone marrow (BM) disorders comprise heterogeneous entities associated with peripheral cytopenias and decreased production of hematopoietic cells in BM. This study was undertaken to analyze immunohistochemical expression of CD34, CD117, and p53 in morphologically diagnosed patients of hypocellular BM (aplastic anemia [AA], hypocellular myelodysplastic syndrome [h-MDS], and hypocellular acute myeloid leukemia [h-AML]). Materials and Methods BM specimens were obtained from patients presenting with pancytopenia/bicytopenia. On 30 patients diagnosed as hypocellular BM, immunohistochemistry (IHC) for CD34, CD117, and p53 was performed. Results BM cellularity was < 30% in all (100%) patients. Blast count was increased in h-MDS and h-AML. Features of dysplasia were noted in six (20%) patients. Out of these, three patients were diagnosed as h-MDS having bilineage/trilineage dysplasia, and the other three patients were of AA (11.5% patients) displaying only dyserythropoiesis. On IHC, percentage of BM CD34+ cells was increased in h-MDS+ h-AML (3.87 ± 0.86) as compared with AA (0.19 ± 0.15) and controls (0.81 ± 0.21), p = 0.01. Percentage of BM p53+ cells was also increased in h-MDS+ h-AML (2.9 ± 2.07) as compared with AA and controls, which did not show any p53+ cells, p = 0.0. No statistically significant difference was observed in the expression of CD117 in h-MDS+ h-AML (4.95 ± 3.40) compared with AA (4.49 ± 1.07), p = 0.99. Conclusion The study demonstrates the usefulness of CD34 and p53 immunoexpression as an important ancillary method in distinguishing various hypocellular BM disorders, especially h-MDS and AA. However, the role of CD117 remains unclear and needs to be evaluated further by larger studies.

Primary leiomyosarcoma of the thyroid gland.

Sarcoma is a rare tumor of the thyroid gland, primary thyroid leiomyosarcoma (LMS) being even rarer. We present a case of LMS of the thyroid in a middle-aged female. Histopathologic examination in conjunction with immunohistochemistry helped to clinch the diagnosis. Knowledge of this entity is important to distinguish it from anaplastic thyroid carcinoma (ATC) and other sarcomas arising in the thyroid and adjacent soft tissue. The prognosis of thyroid LMS is dismal with an extremely poor survival rate.

Evaluation of DNA ploidy and S-phase fraction in fine needle aspirates from breast carcinoma.

BACKGROUND: The use of fine-needle aspiration (FNA) as a primary tool in the diagnosis of breast carcinoma provides opportunity for early proliferative characterization of the tumor. This study was undertaken to assess DNA ploidy and S-phase (SPF) fraction by flow cytometry in fine needle aspirates of patients with breast cancer. METHOD: Fifty patients of breast cancer diagnosed on fine needle aspiration cytology (FNAC) and who subsequently underwent either mastectomy or lumpectomy were included. Material obtained by FNAC was subjected to DNA ploidy and SPF analysis. Immunohistochemical estimation of Ki-67 was done on histopathology sections. The proliferation markers (SPF and Ki-67) were compared with each other and with the histopathologic parameters. RESULTS: On DNA flow cytometry, 27 (54%) cases were aneuploid and 23 (46%) cases were diploid. The median SPF was 12.43% and 4.03% in aneuploid and diploid tumors respectively. Median Ki-67 among aneuploid tumors was 28.6% compared to 8.7% among diploid tumors. Aneuploid tumors were significantly associated with higher values of SPF and Ki-67, with Kappa 0.437 and agreement of 72%. Diploid tumors showed lower values of SPF and Ki-67, with Kappa 0.455 and agreement of 72.7%. Correlation among SPF and Ki-67 was highly significant with Kappa value 0.446, P value of .002 and agreement of 72.3%. CONCLUSION: DNA ploidy and proliferative activity by flow cytometric SPF estimation on fine needle aspirates from breast cancer can provide valuable prognostic and predictive information at the time of diagnosis in patients with breast cancer. This might help in selection of appropriate treatment modality.

Evaluation of Angiogenesis in Multiple Myeloma by VEGF Immunoexpression and Microvessel Density.

Background Multiple myeloma (MM) is a plasma cell disorder characterized by monoclonal proliferation of plasma cells in bone marrow. Plasmablastic MM is a morphologic subset of MM, containing ≥2% plasmablasts of all plasma cells. Methods The study included 30 consecutively diagnosed patients of MM (6 plasmablastic, 24 nonplasmablastic) over a span of 2 years. Angiogenesis in MM was assessed by analysis of vascular endothelial growth factor (VEGF) immunoexpression by plasma cells and microvessel density (MVD) using anti-CD34 antibody. CD34 and VEGF immunohistochemical staining was performed in all the 30 cases. Angiogenesis was studied in relation to plasmablastic morphology and clinical profile to determine if any correlation exists between these. Results The mean VEGF expression of 80.83 ± 7.36 in plasmablastic myeloma cases was significantly higher compared with a mean VEGF of 53.54 ±17.09 in nonplasmablastic cases. Most of the cases (66.6%) of plasmablastic myeloma exhibited strong (3+) VEGF expression. The difference in mean VEGF expression between plasmablastic and nonplasmablastic cases was found to be statistically significant ( p = 0.001). The mean MVD in plasmablastic cases was 44.8 ± 3.69, while in the nonplasmablastic category, the mean MVD was 23.7 ± 5.14, difference being statistically significant ( p < 0.05). Also, a positive correlation was found between VEGF expression and MVD. Conclusion A moderate/strong VEGF intensity and higher MVD were found in cases of plasmablastic MM, suggesting that a more aggressive histological disease may be associated with increased production of VEGF. This finding might be helpful to identify a subset of patients with adverse prognosis and to provide antiangiogenic therapy to improve their survival. However, studies comprising larger number of patients are required to bring out a statistical significance to further substantiate these findings.

Spindle cell lipoma mimicking pleomorphic adenoma: A diagnostic challenge on cytology.

Spindle cell lipoma (SCL) is a benign neoplasm of mature adipose tissue mostly located in the neck, back, and shoulder region. It is composed of a variable admixture of adipocytes, benign spindle cells, and collagen fibres. While the histopathologic features of SCL are quite characteristic and well described, a cytologic diagnosis of SCL might be challenging. We present a case of a 36-year-old female with a swelling in the left preauricular region since last 6 mo. On fine needle aspiration, a diagnosis of lipomatous pleomorphic adenoma (PA) was rendered because of the presence of benign spindle-shaped cells, adipocytes, and abundant myxoid material. However, on histopathologic examination, the diagnosis was SCL which was confirmed by immunohistochemistry. Preauricular region is a rare site of occurrence of SCL. An acquaintance with its varied cytomorphologic features and a knowledge of its unusual sites of occurrence is essential to prevent its misdiagnosis as other myxoid benign and malignant spindle cell lesions.

Rumen-protected B vitamin complex supplementation during the transition period and early lactation alters endometrium mRNA expression on day 14 of gestation in lactating dairy cows.

Greater metabolic demands in high-producing dairy cows are believed to be a cause of sub-fertility in these animals. Previously, supplementation with vitamin B complex molecules has shown benefits in improving milk production, health, and reproductive efficiency of dairy cows. The primary aim of this project was to determine the effects of rumen-protected vitamin B complex supplementation of 100 g of Transition VB (Jefo, St. Hyacinthe, QC, Canada) and 4 g of Lactation VB (VB; Jefo), during the transition and early lactation periods, respectively, compared with a control diet containing no supplementation on d 14 endometrial outcomes of pregnancy. In the vitamin B supplemented cows, we expect to see a change in the mark-up of endometrial genes important for embryo survival before implantation. Multiparous Holstein cows were enrolled into the study 3 wk before parturition and were randomly assigned to either the VB or control treatment. Twice-a-week blood samples, weekly milk samples, and daily feed intake were collected. Cows were enrolled onto a double-ovsynch protocol at 33 ± 3 d postpartum and inseminated by timed artificial insemination. Milk production and components, concentrations of BHB, haptoglobin, and progesterone in serum, and ovarian dynamics were also measured, but no treatment effect was observed. The uterus was flushed on d 14 after artificial insemination (around 72 DIM) for conceptus collection, and endometrial samples were collected at the same time. Overall, 42 cows were flushed and 13 embryos were collected. Analysis of mRNA expression of genes related to embryo development, immune system, adhesion, and regulation of vitamin B molecules showed that OXTR, MUC5B, MUC1, IL1B, SPP, TRD, FZD8, and FOLR1 genes were significantly upregulated in the VB group. Vitamin B supplementation had no effect on the size of the embryo and ovulatory follicle or corpus luteum diameter at embryo collection. In conclusion, the benefits of strategic dietary VB supplementation during the transition and early lactation might be directly linked to endometrial functions required for embryo survival during the peri-implantation period.

Evaluating the Unusual Histological Aspects of Granuloma Annulare: A Study of 30 Cases.

BACKGROUND: Granuloma annulare (GA) is an uncommon dermatologic disorder that presents as annular, skin-colored to erythematous plaques. Histopathologically, it is characterized by palisaded histiocytic granulomas. A definitive diagnosis of GA is based on clinicopathologic correlation. OBJECTIVE: The aim of this study was to study the histomorphologic spectrum of GA. MATERIALS AND METHODS: A total of 30 cases reported as GA over 6 years (2012-2017) were retrieved. The detailed clinical profile and histomorphologic findings on the skin biospies were reviewed. RESULTS: Majority of the cases (40%) presented in the 6th decade of life with a mean age of 48.3 ± 16.5 years and with a female predominance (77%). The lesions were localized in 22 cases (73%). Asymptomatic to erythematous, annular plaques was the most frequent presentation (60%). GA was not suspected clinically in two cases. Histopathologically, interstitial pattern of infiltrate was most common (44%), whereas granuloma formation and palisaded histiocytes were seen in 4 (13%) and 3 cases (10%), respectively. A mixed pattern was observed in 10 (33%) cases. Collagen degeneration was universal finding (100%) and presence of dermal mucin was noted in 24 cases (80%), both of which were important clues to the diagnosis of GA. Additional features such as presence of plasma cells, eosinophils, and vasculitis were noted in 10 (33%), 6 (20%), and 6 (20%) cases, respectively. CONCLUSION: The diagnosis of GA may be challenging owing to its diverse morphology. Acquaintance with the varied histomorphology of GA is of utmost importance to render a correct diagnosis and understand the pathogenesis.

Effect of estrous expression on timing and failure of ovulation of Holstein dairy cows using automated activity monitors.

The aim of this study was to determine if estrous expression, as measured by an automated activity monitor (AAM), affects timing and failure of ovulation of lactating Holstein dairy cows. Cows were equipped with 2 AAM, 1 neck-mounted (AAMC) and 1 leg-mounted (AAML), by 10 d postpartum and enrolled into the trial when their activity crossed the alert threshold on the AAMC. A total of 850 episodes of estrus from 293 different cows were used for this study. When cows were enrolled, their ovaries were scanned by transrectal ultrasonography and gait and body condition scored. Ovaries of cows detected in estrus were scanned twice daily for a maximum of 3 d to determine the disappearance of the preovulatory follicle (ovulation) and the interval from estrus to ovulation was calculated. Physical activity data recorded from the AAM were used to determine estrus behavior using 2 traits: (1) peak activity and (2) duration. Peak activity was only available for the AAML. Peak activity was defined as the maximum activity during an estrus episode. Duration of estrus was defined as the time the activity of the cow exceeded threshold values set by the AAM software. The AAMC correctly identified 87.8% of the estrus alerts, with 12.2% false positives. The average (±standard deviation) intervals from activity alert to ovulation were 25.8 ± 10.2 and 24.7 ± 9.3 h for the AAMC and AAML, respectively. Changes in estrous expression were associated with differences in the interval from alert to ovulation. Cows with short intervals to ovulation were found to have less intense estrous expression than cows with medium and long length intervals to ovulation using the AAMC, whereas using the AAML, cows with short intervals to ovulation exhibited less intense estrous expression than cows with medium but the same as those with long intervals to ovulation. Furthermore, irrespective of the AAM, estrus events with less estrous expression had increased odds of having a short interval to ovulation (below the median of 20 h) when compared with those having greater estrous expression (2.6 and 1.9 increased odds for the AAMC and AAML, respectively). Ovulation failure was affected by estrous expression because estrus events with greater peak activity or longer duration had reduced ovulation failure compared with those with less estrous expression (AAMC peak activity: 1.9 ± 1.4 vs. 9.5 ± 1.7%; AAML peak activity: 2.3 ± 1.4 vs. 6.2 ± 1.5%; AAMC duration: 2.1 ± 1.4 vs. 8.9 ± 1.7%). In addition, cows with more estrous expression had greater pregnancy per artificial insemination than those with less estrous expression with both the AAMC (42.3 ± 0.4 vs. 31.7 ± 0.4%) and the AAML (43.1 ± 0.4 vs. 36.3 ± 0.4%). Pregnancy per artificial insemination results were consistent even when removing cows that failed to ovulate. In conclusion, expression of estrus was highly associated with ovulation timing, ovulation failure, and fertility when using 2 different AAM. Cows with greater estrous expression have longer intervals from activity alert to ovulation, experience less ovulation failure, and have greater pregnancy per artificial insemination.

Effect of metritis on endometrium tissue transcriptome during puerperium in Holstein lactating cows.

The objective of this prospective cohort study was to evaluate the effect of parity and uterine health status postpartum on the gene expression profile of the endometrium early post-partum. Twenty-four Holstein cows were randomly selected (16 multiparous (MP) and 8 primiparous (PP)) and endometrium biopsies were collected on days 1, 3, and 6 after calving and clinically monitored for metritis. Rectal temperature was measured twice and fever was defined as a temperature ≥39.5 °C. A case of metritis was diagnosed with the presence of red-brown watery, foul-smelling uterine discharge or a purulent discharge with more than 50% pus and fever between days 1 and 6 postpartum. Cows were then retrospectively selected (cows diagnosed with metritis were paired with healthy ones) to analyze the expression of 66 genes measured on the NanoString nCounter Analysis System. The genes selected were related with adhesion, immune system, steroid and prostaglandin biosynthesis regulation, insulin metabolism and transcription factors, and nutrient transporters. The results indicated a different pattern on genes related to immune function by parity. PTX3, involved in antigen presentation, was increased in healthy MP compared with healthy PP whereas inflammatory cytokine TNFα and complement-related protein SERPING1 was upregulated in MP compared with PP (P < 0.05). As expected, presence of a metritis condition affected the expression of genes related to immune function. There was an increased expression of the antiviral factor MX2 and MYH10 gene, which is involved in macrophages recruitment, in metritic compared with healthy cows (P < 0.05). Differences in uterine involution from cows diagnosed with metritis were reflected by the downregulation of IGF1 (P < 0.10), involved in endometrium remodeling, and a possible compensatory upregulation of its receptor IGFR1 (P < 0.05). A greater expression of prostaglandins and oxytocin receptors (PGR and OXTR), involved in the involution process, were observed in metritic PP compared with healthy PP (P < 0.05). Overall, it seems that metritis significantly modulate processes closely tied with the physical involution of the uterus early post-partum (IGF1, IGFR1, PGR, OXTR), whereas both metritis and multiparous cows tended to upregulate genes related to immune response (PTX3, TNFα, SERPING1, MX2, MYH10).

Papillary Pattern in Acinic Cell Carcinoma of Parotid Gland: A Potential Diagnostic Pitfall on FNAC.

Fine needle aspiration cytology (FNAC) is often the first investigation in the work up of salivary gland lesions. However, its diagnostic accuracy is limited by the high rates of false positives and false negatives. Usually, acinic cell carcinoma is prone to be underdiagnosed because of the cytological similarity of the tumor cells to normal acinar cells, however rarely, a predominant papillary architecture on cytology may cause confusion with adenocarcinomas. We present a case of a 45-year-old male with a painful swelling of the right parotid region. FNA smears revealed a predominant papillary architecture and focal acinar pattern. A provisional diagnosis of acinic cell carcinoma was given, which was confirmed on histopathology. Familiarity with the cytomorphologic features of acinic cell carcinoma is cardinal for an accurate diagnosis and appropriate management.

Periampullary Carcinoma: Unusual Sites of Metastasis.

BACKGROUND: Periampullary neoplasms include carcinomas of the duodenum, ampulla of Vater, distal common bile duct, and pancreas. The aggressive course of these neoplasms is attributable to the delay in diagnosis, as patients have no symptoms until advanced stages of the disease. More than half of patients have distant metastasis at the time of diagnosis. The most frequent sites of metastasis are the liver, lymph nodes, peritoneum, lung, bone, kidney, and, rarely the skin. CASE REPORT: We report the case of a 45-year-old female patient with adenocarcinoma of the ampulla of Vater metastasizing to the right parietal skull with overlying cutaneous involvement 1 year after a Whipple procedure. CONCLUSION: Cutaneous and skull metastasis of periampullary neoplasms is unusual, but early recognition of such metastatic disease is important because it indicates a poor prognosis for the patient.

Primary Leiomyosarcoma of the Mesentery: A Case Report.

Mesenteric masses are infrequent lesions ranging from benign cysts to aggressive malignancies and often present a diagnostic and therapeutic challenge. The mesentery is a frequent recipient of metastases from the gastrointestinal tract, pancreas, and biliary cancers. Primary mesenteric tumors are relatively rare, mostly mesenchymal in origin and benign in nature. Examples include gastrointestinal stromal tumors and smooth muscle tumors. We describe a 50-year-old woman, who presented with a lump in the left hypochondrium along with altered bowel habits of 2 years' duration. Imaging revealed a cystic lesion, raising the differentials of a benign lesion. Exploratory laparotomy revealed a large cystic mass in the mesentery closely abutting the jejunal loops. This was followed by mass resection along with a segment of the jejunum. Histopathological examination revealed features of a leiomyosarcoma. Postoperatively, the patient developed a colocutaneous fistula, which was re-excised. At 6 months' follow-up, the patient is doing well. Our case demonstrates the diagnostic challenge posed by the atypical clinical and imaging features of this tumor at an unusual site.

Inflammatory Myofibroblastic Tumor of the Breast Mimicking Malignancy in an Elderly Male.

BACKGROUND: Inflammatory myofibroblastic tumor (IMT) is a rare, distinctive lesion composed of a proliferation of myofibroblastic spindle cells accompanied by an inflammatory infiltrate. It was first described in the lung, but its occurrence at various extrapulmonary sites has also been reported. The literature mentions only a handful of cases of IMT in the breast and only 1 case in the male breast. We report the second case of IMT in the male breast. CASE REPORT: A 60-year-old male presented with a large, lobulated lump in the left breast that had progressively increased in size during the past year. The lump measured 15 × 10 cm. Ultrasonography revealed a solid mass lesion with regular borders in the subcutaneous plane of the left anterior chest wall. Fine-needle aspiration cytology showed a cellular mesenchymal tumor. Macroscopically, the nodule was firm, circumscribed, and yellow. On microscopic examination, the tumor was composed of bland spindle cells arranged in sheets and short fascicles along with a rich inflammatory infiltrate comprising predominantly plasma cells, admixed with lymphocytes, neutrophils, and eosinophils. On immunohistochemistry, the tumor cells were positive for vimentin, focally positive for smooth muscle antigen, and negative for anaplastic lymphoma kinase, CD34, S100, ß-catenin, and cytokeratin. Thus, a final diagnosis of IMT was rendered. CONCLUSION: IMT is a rare entity with intermediate clinical behavior. Knowledge of this entity and its recurrence and metastatic potential is of paramount significance to guide appropriate treatment and follow-up.

Bilateral rhabdoid meningioma mimicking glioma: an unusual occurrence.

Rhabdoid meningioma is an infrequent variant of meningioma, introduced for the first time in the 2000 World Health Organization's classification of tumors of the nervous system. Owing to its aggressive clinical course and high proliferative index, it has been classified as a grade III neoplasm. We describe a fifty-year-old male with headache, weakness of limbs, and altered sensorium. CT showed hetero-dense enhancing mass lesions in both right and left parietal areas raising suspicion of high grade glioma. Histopathologic and immunohistochemical examination revealed a tumor with features of rhabdoid meningioma. A review of literature did not reveal any bilateral occurrence of this tumor.

Pleomorphic Sarcoma of Breast: A Report of Two Cases and Review of Literature.

Sarcomas account for less than 1% of all primary breast malignancies, pleomorphic sarcoma of the breast being even rarer. We present two cases of pleomorphic sarcoma of the breast in a 35-year-old and a 43-year-old female. An extensive review of the available literature with evaluation of the etiology, changing terminologies and histopathologic features of pleomorphic sarcoma of the breast are discussed. The prognostic factors and treatment modalities have also been reviewed.

Papillary carcinoma thyroid with anastomosing channels: An unusual morphology.

Papillary thyroid carcinoma (PTC) is the most common thyroid malignancy, accounting for 70%-80% of all thyroid malignancies. It is biologically indolent and has an excellent prognosis. Variations in histopathologic patterns are known to influence prognosis and often result in a diagnostic dilemma. We report an unusual case of a 35-year-old female with papillary carcinoma of a thyroid isthmus showing anastomosing channels on histopathology, a distinctive pattern that has not been described in PTC. Similar to tumor cells, the lining cells of these channels were also positive for thyroid transcription factor 1, thyroglobulin, and cytokeratin-19 and negative for CD34 and CD31. The diagnosis of PTC should rely on nuclear morphology rather than architecture. Pathologists should be aware of different variants because some of these variants show aggressive behavior and poor outcome. The present report highlights the distinctive pattern of PTC, recognition of which is important to avoid any diagnostic pitfall.

Myeloid sarcoma: a report of four cases at unusual sites.

Background: Myeloid sarcoma (MS) or granulocytic sarcoma is a rare tumor consisting of myeloid blasts with or without maturation occurring at anatomic sites other than the bone marrow. MS can involve any organ system but shows a predilection for skin, bone, and soft tissues of head and neck region. Case report: We report four cases of MS occurring at unusual sites, out of which three were de novo and one was associated with acute myeloid leukemia (AML). Conclusion: Although MS is associated with AML, it can rarely present without any existent hematologic disease. Differential diagnosis of a soft tissue mass should include MS even in the absence of leukemia. Establishment of the correct diagnosis depends on morphologic, histochemical, and immunohistochemical examination.