Immunoperoxidase Staining Research Articles

SUN-394 RENAL FUNCTION CORRELATION WITH HISTOLOGIC ACTIVITY AND CHRONICITY INDICES IN ADULT PATIENTS WITH GLOMERULONEPHRITIDES AT OBAFEMI AWOLOWO UNIVERSITY TEACHING HOSPITALS COMPLEX ILE-IFE

Glomerulonephritis (GN) is one of the major causes of chronic kidney disease (CKD) worldwide. It remains the leading cause of End Stage Kidney Disease (ESRD) in many developing countries including Nigeria with varying clinical course and histologic pattern. The clinical outcome and the degree of renal impairment observed in patients with glomerulonephritides is dependent on the type and extent of the histological lesion identified in the glomerulus and tubulo-interstitium. Renal biopsy remains an important diagnostic tool in patients with glomerulonephritis with varying indications. This study is unique because the histologic features observed in the glomerulus and tubulo-interstitium were graded, scored and correlated with clinical as well as biochemical features. This will add to the body of knowledge on glomerulonephritides in this environment. The objectives of the study was to determine the grading and severity of histologic features observed in the glomerulus and tubulo-interstitium and to determine the relationship (if any) between clinical, biochemical and histologic features in adults with GN at OAUTHC, Ile-Ife. The study was a cross sectional hospital based study of seventy (70) consecutive adult patients with features of glomerulonephritis who presented at the nephrology and other clinics of OAUTHC Ile-Ife. Renal function was assessed and renal biopsy performed after obtaining written informed consent. The renal tissues obtained were subjected to light microscopy and immunoperoxidase staining with IgA, IgM, IgG and C3 antibodies and the degree of involvement of glomeruli, tubules, interstitium and vessels were graded from 1 to 5 for the glomeruli and 1 to 3 for the tubulo-interstitium respectively according to severity. Total activity and total chronicity indices were collated and their association/correlation with clinical parameters assessed. A total of seventy patients participated and completed the study. Three (3) patients had inadequate renal tissue for histologic diagnosis hence data analysis was based on the remaining sixty seven (67) patients. Statistically significant correlations were found between the interstitial scores for activity (interstitial oedema, interstitial infiltrate) with serum creatinine and GFR: interstitial oedema with serum creatinine (r=0.35, p=0.003) and GFR (r=-0.38, p=0.004); interstitial infiltrate with serum creatinine (r=0.52, p<0.0002) and GFR (r=-0.70, p=0.002) and total activity index with serum creatinine (r=0.60, p=0.0001) and GFR (r= -0.48, p = 0.004). Statistically significant correlations was also seen between total chronicity index and test of renal function: total chronicity index with serum creatinine (r=0.62, p=0.001) and GFR (r= -0.58, p<0.00001). Both activity and chronicity indices significantly influence renal function. The higher they are the lower the GFR and vice versa, this suggests aggressive management protocol for patients with higher indices. The histologic scoring system would assist in assessing the severity of the lesion which could influence renal disease progression and management plans.

SUN-395 CLINICO-PATHOLOGIC PATTERN OF ADULT GLOMERULONEPHRITIDES IN NIGERIA: A SINGLE CENTRE STUDY

Glomerulonephritis (GN) is one of the major causes of chronic kidney disease (CKD) worldwide. It remains the leading cause of End Stage Kidney Disease (ESRD) in many developing countries. It is presumed to be responsible for more than half of patients with ESRD in Africa. The clinical course, pattern and presentation of glomerular disease varies world-wide and may be due to differences in genetics of underlying population or exposure to putative antigens or agents that induce or trigger these responses. This study evaluated the clinicopathologic patterns of glomerular diseases in a homogenous black population in the hope that it will provide some data on glomerulonephritides in this population. The objective of the study was to describe the pattern of biochemical abnormalities and histopathologic variants in adults with glomerulonephritis at OAUTHC, Ile Ife. The study was a cross sectional hospital based study of seventy (70) consecutive adult patients with features of glomerulonephritis who presented for check up at the nephrology and other MOPD clinics, GOPD as well as the hospital staff clinic of OAUTHC Ile-Ife. A structured 4-part proforma was drawn up and administered to obtain information on socio-demographic, clinical, anthropometric parameters and laboratory investigation results. Relevant laboratory tests and investigations were performed. Renal function was assessed and renal biopsy performed after obtaining written informed consent. The renal tissues obtained were subjected to light microscopy and immunoperoxidase staining with IgA, IgM, IgG and C3 antibodies and the results interpreted. A total of seventy patients participated and completed the study. Three (3) patients had inadequate renal tissue for histologic appraisal hence data analysis was based on the remaining sixty seven (67) patients. The age range of the study population was between 18 and 65 years with a mean age of 28.43 (±10.33) years. Forty- five males (67.2%) and twenty-two females (32.8%) were in the study population with an overall male to female ratio of 2:1. The most common clinical manifestation was nephrotic syndrome which was present in 41 (61.2%) of the study population. Nephrotic syndrome was followed by nephritic-nephrotic syndrome accounting for 20.9% of the clinical syndrome with nephritic syndrome, AUA and macroscopic haematuria accounting for 13.4%, 3.0% and 1.5% respectively. The most common histological diagnosis was FSGS seen in 30 patients (44.8%). Focal segmental glomerulosclerosis (FSGS) was the commonest histological type found in these patients with glomerulonephritis, while the commonest clinical presentation was nephrotic syndrome.

Characterization of B1-cells during experimental leukomogenesis.

Bovine leukemia causes a significant polyclonal expansion of CD5+, IgM+ B lymphocytes, known as persistent lymphocytosis (PL), in approximately 30% of infected cattle. However, it is not yet clear what happens to this subpopulation of B cells in the early period of infection of animals. Quantitative characterization of IgM+ and CD5+ B cells during the immune response, which can provide important information on the mechanisms of lymphocyte priming in BLV infection. The experiment used BLV-negative calves of black-motley breed at the age of 8 months (n = 11). Animals (n = 8) were intravenously injected with blood of a BLV-positive cow. Control calves (n = 3) were injected with saline. Studies were performed before and after infection on days 5, 7, 14, 21, 28 and 65 of the immune response. The determination of the number of B-lymphocytes in the blood was carried out by the method of immunoperoxidase staining based on monoclonal antibodies to IgM, CD5. As a result of the studies, it was found that the level of CD5+ B cells increases on the 14th day of the primary immune response, characterized by polyclonal proliferation of CD5+ B cells, which are the primary target for BLV. Our research data confirm that in the lymphocytes of experimentally infected cattle, surface aggregation of IgM and CD5 molecules on B-lymphocytes is absent. It is known that the wave-like nature of IgM synthesis, which was shown in previous studies, depends on a subpopulation of B1 cells. After 7 days of the immune response, IgM+ and CD5+ cells do not correlate, which shows their functional difference. The increase in CD5+ cells is probably not associated with B cells, but with T cells differentiating under the influence of the virus. A subset of B1 cells is the primary target of cattle leukemia virus. The 65th day of the immune response is characterized by the expansion of IgM+ B cells, a decrease in the number of CD5+ cells and a uniform distribution of receptors around the perimeter of the cells.

Susceptibility of Chicken Embryos, Sheep, Cattle, Pigs, and Chickens to Zika Virus Infection.

The susceptibility of sheep, cattle, pigs, chickens and chicken embryos to Zika virus infection was evaluated by experimental inoculation with Zika virus Thailand strain isolated from a Canadian traveler in 2013. The inoculated animals did not develop any clinical signs of disease nor evidence of Zika virus replication in peripheral blood, cerebrospinal fluid and tissues including brain and spinal cord assessed by real-time RT-PCR. Sera were also negative for Zika virus antibodies by Zika virus neutralization assays as well as Zika virus immunoperoxidase staining of Zika infected Vero cells. Chicken embryos were inoculated by different routes including yolk sac (4 day old embryos), chorioallantoic membrane (8 day old embryos), amniotic fluid (8 day old embryos) and intravenous routes (12 day old embryos). Virus replication in chicken embryos was observed in the brain and body tissues following intravenous (IV), yolk sac (YS), chorioallantoic membrane (CAM), and amniotic fluid (AF) inoculation routes. The highest mortality was observed in embryos inoculated via yolk sac. The dead embryos showed diffuse muscular hemorrhages. The yolk sac inoculated chicken embryos showed delayed hatching and displayed neurological signs immediately after hatching. These studies demonstrate that 8 week old sheep, 6 month old cattle, 4 week old pigs, and 4 week old chickens are not susceptible to Zika virus infection when inoculated experimentally and therefore unlikely to pose a risk as Zika virus reservoirs. However, chicken embryos are highly susceptible to Zika virus resulting in clinical disease of chicks after hatching. This study demonstrates that Zika virus has a tropism for embryonic tissue and that chicken embryos can be used as a model to study Zika virus replication and pathogenesis.

A Melanin bleaching method to prevent non-specific immunostaining of chicken feathers

Melanin in pigmented organs like the skin is known to react with 3,3′-diaminobenzidine (DAB) to give a brown colour indistinguishable from the colour that DAB imparts to target antibodies bound to specific antigens. This can lead to false positives in chicken feathers during immunoperoxidase staining. Here, we present a simple, fast and practical method for bleaching chicken feathers which can be applied prior to immunohistochemistry staining without affecting specific antigen-antibody binding. To our knowledge, this is the first report of a melanin-bleaching technique prior to immunoperoxidase staining techniques of chicken feathers for detection of pathogens. Optimisations of the method include:•Removal of melanin from tissue sections using a short incubation with potassium permanganate followed by incubation with oxalic acid prior to immunostaining for improved specificity.•This technique did not affect the antigenicity of infectious laryngotracheitis virus antigen and did not cause damage or detachment of tissues from the slides.

127 Chemokine receptor 2 (CCR2) is expressed in growing oocytes, and its deficiency affects follicular activation and long-term female fertility in mice

Chemokine receptor 2 (CCR2) has important functions in several biological processes, including activation of PI3K/Akt/mTOR signalling, a key pathway in follicular activation. However, there is no report about the role of CCR2 in ovarian follicular physiology. The objectives of this study were (1) to immunolocalize CCR2 in mice ovaries and (2) to evaluate the effects of CCR2 deficiency on follicular growth during adult life and aging. A total of 74 C57Bl/6 (wild type, WT) and 68 B6.129S4-Ccr2tm1Ifc/J (CCR2−/− (knockout)) female mice were used. For objective 1, ovaries were collected from WT mice at 1.5 months old (m.o.), fixed in 4% PFA, embedded in paraffin, and used for immunoperoxidase staining with an anti-CCR2 antibody [EPR19698] (ab222496; 1:200) and using anti-rabbit IgG (Ab6721, 1:100) as a secondary antibody. Also, MII oocytes from oviducts of superovulated WT mice were processed with the same primary antibody for immunofluorescence. For objective 2, body and ovarian weight were evaluated. Follicle populations were assessed in WT and CCR2−/− mice at 1.5, 2.5, 6, 10, and 12m.o., by serial sectioning; the total follicle population was counted in every third section in the whole ovary. Additionally, ovarian total RNA isolation was performed from WT and CCR2−/−. Real-time PCR was used to evaluate differential gene expression according to standard protocols, using primers for Bax, Casp3, Bcl2, Fshr, and B-actin (endogenous control). The data were analysed using the GraphPad Prism Software, using t-test, and a P-value of 0.05 was considered as significant. Localization of CCR2 was observed exclusively on the membrane and cytoplasm of growing oocytes in primary, secondary, antral, and atretic follicles, as well as on ovulated MII oocytes membrane and cytoplasm. Body and ovarian weight were similar between WT and CCR2−/− mice. At 1.5m.o., CCR2−/− mice had more primordial follicles and fewer primary and secondary follicles compared with WT (P&amp;lt;0.05), whereas there was no difference in the antral follicle populations. Follicular activation (primordial to primary transition) and atresia rates were decreased in CCR2−/− (P&amp;lt;0.05) at 1.5m.o. A downregulation of pro-apoptotic genes (Bax and Casp3) was observed on CCR2−/− (P&amp;lt;0.05), while anti-apoptotic Bcl2 was upregulated (P&amp;lt;0.05) compared with WT in 1.5-m.o. animals. A larger ovarian follicular reserve at 1.5, 2.5, and 6m.o., but not at 10 or 12m.o., was observed in CCR2−/− mice. At 6 to 12m.o., CCR2−/− ovulated more (P&amp;lt;0.05) MII oocytes than WT. Altogether, these data may suggest that CCR2 plays an important role in the regulation of ovarian follicular reserve mobilization, potentially affecting reproductive lifespan. Financial support was provided by Fapemig and CNPq.

AN ATYPICAL PRESENTATION OF CYTOMEGALOVIRUS PNEUMONIA

SESSION TITLE: Monday Medical Student/Resident Case Report Posters SESSION TYPE: Med Student/Res Case Rep Postr PRESENTED ON: 10/21/2019 02:30 PM - 03:15 PM INTRODUCTION: Cytomegalovirus (CMV) is a viral pathogen that is seen in 40-100% of adults by the fourth decade (1). Infection in an immunocompetent individual is usually subclinical, with severe life-threatening infection seen in immunocomprised individuals. Steroid use has a weak association for infection reactivation in an immunocompetent host. We present the case of a chronic kidney disease (CKD) patient on steroids who was diagnosed and treated for CMV interstitial pneumonia. CASE PRESENTATION: 78-year-old female with a past medical history of interstitial lung disease, idiopathic thrombocytopenia purpura on steroids, CKD stage III who presented from a nursing facility where she was recuperating from a recent hospitalization due to pneumonia. During her prior admission, she was treated with steroids, vancomycin and cefepime for a presumed bacterial pneumonia. Patient presented with shortness of breath. Vital signs were notable for a temperature of 98.4 F, heart rate 102 bmp, RR 24/min, and SaO2 90% on 7L NC. Physical exam revealed bibasilar rales. Labs showed a white blood cell count of 10.1 Thou/uL, creatinine of 1.5 mg/dL, lactic acid 2.8 mmol/L, procalcitonin of 0.12 ng/mL, lactate dehydrogenase (LDH) of 544 U/L. Computed tomography (CT) of the chest showed increasing ground glass changes compared to previous admission imaging. Patient was started on steriods, vancomycin and cefepime. Given worsening radiographical findings, history of ILD, and elevated LDH in the setting of chronic steroids, there was a suspicion for pneumocystis pneumonia. She underwent a video-assisted thoracoscopic surgery lung biopsy, bronchoalveolar lavage and wedge resection of the right lung. Pathology showed microscopic honey combing and extensive subpleural fibrosis, consistent with interstitial pneumonia. CMV cytopathic effect was confirmed with immunoperoxidase staining. PCR testing and tissue culture also returned positive for CMV; viral load was 81,581 IU/mL. Patient was started on a six-week course of valganciclovir with a plan to taper steroids as an outpatient. Patient was unfortunately lost to follow up thereafter. DISCUSSION: Severe life threatening CMV infection is seen in immunocomprised risk groups particularly transplant recipients, patients affected with AIDS, malignancy and neonates. Steroid use has a weak association with CMV reactivation; CKD also has been postulated as a risk factor for reactivation due to malfunctioning white cells. CONCLUSIONS: Although CMV pneumonia targets a subtype of immunosuppressed patients, it should be considered a differential in patient’s on chronic steroid treatment and CKD. This case emphasizes the importance of having a high degree of suspicion of CMV pneumonia in a patient on chronic high dose steroids, in clinical setting, such as this one. Without the correct diagnosis, obtained by PCR of a BAL sample, treatment opportunities will be missed, leading to poor outcomes. Reference #1: Grilli, Elisabetta, et al. “Cytomegalovirus Pneumonia in Immunocompetent Host: Case Report and Literature Review.” Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology, U.S. National Library of Medicine, Dec. 2012. Reference #2: Al-Omari, A., Aljamaan, F., Alhazzani, W., Salih, S., & Arabi, Y. (2016). Cytomegalovirus infection in immunocompetent critically ill adults: literature review. Annals of intensive care, 6(1), 110 Reference #3: Carpani, G., Foresti, S., Dell'Oro, R., Grassi, G., & Bombelli, M. (2019). Severe systemic cytomegalovirus infection in an immunocompetent patient outside the intensive care unit: a case report. BMC infectious diseases, 19(1), 34. https://doi.org/10.1186/s12879-018-3621-8 DISCLOSURES: No relevant relationships by Antonie Auguste, source=Web Response No relevant relationships by Amreet Aujla, source=Web Response No relevant relationships by Sarah Banks, source=Web Response No relevant relationships by Manmeet Grewal, source=Web Response No relevant relationships by Ann Palmer, source=Web Response

2596 Diagnosis of Terminal Ileum Carcinoid Tumor Despite Prior Screening Colonoscopies—Making the Case for Routine Terminal Ileoscopy During Colon Cancer Surveillance: A Case Report

INTRODUCTION: The term carcinoid is generally applied to well-differentiated neuroendocrine tumors originating in the tubular digestive tract, lungs or rare primary sites. Small bowel neuroendocrine tumors may be asymptomatic at presentation and found incidentally. Among symptomatic patients, 40% experience abdominal pain as an initial symptom. The pain is usually vague, nonspecific and misconstrued to represent irritable bowel syndrome for years before a diagnosis is made. A diagnosis may be delayed, or even missed, due to terminal ileum visualization not being a routine part of screening colonoscopy, a result of the perceived difficulty of the maneuver. We present the case of a female patient who was diagnosed with carcinoid tumor only after she became symptomatic despite having routine colonoscopy screening prior. CASE DESCRIPTION/METHODS: A 74-year-old female without significant past medical history initially presented to clinic with complaints of non-specific abdominal pain and intermittent non-bloody diarrhea. Vital signs and physical exam were unremarkable. The patient subsequently underwent a colonoscopy, during which she was found to have inflamed ileal mucosa. Biopsy was taken and microscopic findings revealed a tumor composed of small uniform cells demonstrating a festoon growth pattern with ovoid nuclei. Immunoperoxidase staining revealed the tumor cells to be positive for pankeratin, chromogranin and synaptophysin, confirming a diagnosis of carcinoid tumor. Patient has since been referred for surgical evaluation. DISCUSSION: Due to the high amount of patients being screened, there is an urgency now more than ever to minimize colonoscope advancement and mucosal inspection time. As depicted in our case, patients may go through more than one surveillance colonoscopy with a missed diagnosis, particularly in the terminal ileum. This is especially a concern in asymptomatic patients with carcinoid tumors. Though evidence has shown improvement in colonic diagnoses, completion of colonoscopy by passage through the ileocecal valve is not yet routinely performed in screening colonoscopies. Difficulty in intubation of the valve as well as an anticipated increase in procedure time have become barriers in establishing this as a standard. With practice, ileoscopy may be highly achievable in routine colonoscopies. This will allow for a decrease in missed diagnoses, early detection and thus more treatable cases overall.

The effects of tramadol on cancer stem cells and metabolic changes in colon carcinoma cells lines

Opioids are widely used in the treatment of cancer related pain. They mainly exert their effects on opioid receptors. The most common opioid in the treatment of pain is morphine. Previous studies show that they may have effects on cancer cell behavior. These may include apoptosis, angiogenesis, invasion, inflammation and immune reactions. Tramadol, also an opioid is widely used in the treatment of cancer pain and is not well studied in cancer behavior. We aimed to investigate the effects of tramadol on cancer stem cells and metabolic changes in colon carcinoma cells.We used Colo320 (ATCC, CCL-220), Colo741 (ECACC, 93052621) and HCT116 (ATCC, CCL-247) colon cancer cell lines. CD133 was considered colon cancer stem cell marker and used to sort CD133+ and CD133− cells by magnetic cell sorting. MTT (mitochondria-targeted therapeutics) technique was used to detect tramadol's cytotoxic effect on cells in the study groups. Cells were treated with 1 mg/kg, 1.5 mg/kg and 2 mg/kg tramadol for 24 h at 37 °C and 5% CO2.Caspase-3, Ki-67, Bcl-2 and VGEF distributions were performed using indirect immunoperoxidase staining for immunohistochemical analysis. The study showed that tramadol has triggering effect on apoptosis in Colo320 colon cancer stem cells.

Chromogranin A Tubulopathy: Differing Histopathologic Patterns of Acute Tubular Injury in the Setting of Neuroendocrine Neoplasms

IntroductionNeoplasms of neuroendocrine derivation or differentiation may express specific peptides, some of which are capable of producing clinical symptomatology and others used as biomarkers: one such peptide being chromogranin A (CGA). Herein, we describe histopathologic changes present in kidney specimens from patients with such neoplasms, and illustrate 2 patterns of acute tubular injury (ATI) attributable to CGA.MethodsEleven patients with a history of a neoplasm of neuroendocrine derivation or differentiation and having histopathologic sampling of the kidney were retrospectively identified, 3 of whom had ATI with either engorgement of the proximal tubular epithelium by resorbed material or tubular cast formation.ResultsTwo patterns of ATI were observed. One characterized by acutely injured proximal tubular cells engorged with resorption granules that expressed CGA via immunoperoxidase staining. Another pattern was characterized by intraluminal tubular cast material associated with ATI that did not exhibit restriction of immunoglobulin light chains (LCs), but immunoperoxidase staining for CGA revealed that the cast material was composed of the neuroendocrine-associated peptide. The level of serum CGA does not appear to necessarily equate to developing either of these 2 patterns of ATI.ConclusionsPatients with a neoplasm of neuroendocrine derivation or differentiation may develop ATI, and in certain cases may be secondary to CGA renal tubular deposition.

Papillary thyroid carcinoma presenting as a primary renal tumor with multiple pulmonary and bone metastases: a case report

BackgroundPapillary thyroid carcinoma is the most common endocrine malignancy. Distant metastasis from differentiated thyroid carcinoma is infrequent and the metastasis rate of papillary thyroid carcinoma is lower than that of follicular thyroid carcinoma. Distant metastases from differentiated thyroid carcinoma are usually seen in the lungs and bones; however, renal metastasis is very rare.Case presentationHere we describe an 85-year-old Caucasian woman who presented with right flank pain 10 years ago. We describe a case of papillary thyroid carcinoma presenting as a primary renal tumor with extensive pulmonary and bone metastases. Abdominal screening with computed tomography revealed a mass on her right kidney, which was considered a primary renal cell carcinoma and she underwent a right nephrectomy. Unexpectedly, papillary thyroid carcinoma metastasis was diagnosed from demonstrative histopathological findings, such as positive immunoperoxidase staining for thyroglobulin. A total thyroidectomy was performed. Unenhanced thoracic computed tomography and skeletal scintigraphy revealed bilateral multiple nodules in her lungs and bone metastasis on T10 vertebra and right sacroiliac joint. Initially, 30 Gy radiotherapy was implemented to her T9–10 vertebrae and then she was treated with a total of 800 mCi radioactive iodine for ablation. A radioactive iodine whole body scan was performed after each 200 mCi and continuous progression was shown in each scan. After she was lost to follow-up for 3 years, she referred to our clinic again with a draining mass on her neck and we planned radiotherapy to this giant mass.ConclusionOur patient was surprisingly still alive after metastatic disease was diagnosed 10 years ago and she had no major complaint other than a draining mass on her neck. Our primary aim by sharing this case is to underline potential renal metastasis from papillary thyroid carcinoma. In other words, when approaching primary renal tumors, possible distant metastases of other organs need to be kept in mind for differential diagnosis. In addition, it should be noted that if managed appropriately, the long-term survival in patients with papillary thyroid carcinoma with multiple organ metastases could be encouraging.

MON-331 Pulmonary Carcinoidosis Presenting with Persistent Pneumothorax

Introduction: Lung neuroendocrine (NET) or carcinoid tumors are a rare group of pulmonary neoplasms characterized by insidious clinical behavior. In this report, we present a case of pneumothorax refractory to conservative management with concomitant pulmonary carcinoidosis. We also discuss the diagnostic and management approach followed in such cases. Case: 80 year-old male with history of asthma and Type 2 DM presented to the ER with worsening cough and shortness of breath, and was found to have a left sided spontaneous pneumothorax. Chest tube was inserted. However, due to persistent leak, PleurX catheter was placed, and patient was discharged home afterwards. However, he presented with subcutaneous emphysema on the same day and was subsequently readmitted. Persistent pneumothorax was managed conservatively. PleurX catheter was attached to underwater seal with negative pressure and respiratory status was optimized given evidence of asthma exacerbation. Treatment with Methylprednisolone IV, Montelukast, Tiotropium, Mometasone Furoate/Formoterol inhalers was initiated. However, air leak persisted despite conservative management for 13 days. CT surgery elected to proceed with VATS. Multiple wedge resections were performed along with partial pleurectomy, lysis of adhesions and decortication of left upper lobe. Pathology revealed a sub-pleural 5-mm carcinoid tumor with solid nests of cells along with lung tissue with emphysematous changes, pleural scarring and bullous formation. No nuclear pleomorphism or mitotic figures were noted. Immunoperoxidase stains were positive for TTF-1, synaptophysin, chromogranin and CD56 and negative for CK5/6, calretinin and P-40. Serum Chromogranin A (CgA) was elevated 431 ng/l. Post-op course was uncomplicated and patient was discharged to a rehab facility. Discussion: Lung NETs account for 1-2% of all lung malignancies in adults and 30% of all NETs. Incidence ranges from 0.2-2/100,000 population per year. While some patients may be asymptomatic, others present with obstructive symptoms due to mass effect. Incidence of pneumothorax as a complication of lung neoplasms is rare (0.05-1.4% of all pneumothoraces). Clinical presentation includes carcinoid syndrome, Cushing's syndrome and acromegaly. Diagnosis is confirmed either by bronchoscopic biopsy (for central lesions) or by transthoracic needle biopsy (for peripheral lesions). For patients with localized lung NETs, surgical resection is the treatment of choice. Lung NETs have an excellent prognosis following surgical resection, with 5-year survival rates of 87-100%. Immunohistochemical identification utilizes tissue markers like synaptophysin, neuron-specific enolase, and chromogranin. Serum CgA is usually elevated and is used as a marker to assess disease progression, response to therapy, or recurrence after surgical resection.

Matrix Fibrous Proteins and Cells in Central Vein Fibrosis of Cadaveric Livers

The central vein (CV) is defined as a hepatic vein at the center of the classic liver lobule. It has a cross‐sectional diameter not exceeding 125 μm. The endothelium lining of the vein is thin and has openings that drain every sinusoid of a liver lobule. The rim or wall of the CV is less than 2 μm in thickness with sparse matrix fibrous proteins and cells. Fibrosis of the CV (CVF), also is called perivenular fibrosis, is marked by thickening of the rim of the vein. The fibrotic tissue deposited in the CV may act like a “pervenular fibrous dam” that obliterates the sinusoidal openings into the vein, resulting in sinusoidal portal hypertension. CVF occurs early in hepatic fibrogenesis of rats induced by CCl4 or porcine serum administration. CVF often accompanies perisinusoidal fibrosis of the centrilobular area in alcoholic liver disease and nonalcoholic fatty liver disease. In the liver of elderly cadavers (mean age, 82.1±10.4), we have observed a 49.2 % incidence (31/63 livers) of CVF, but the matrix components of the fibrotic veins are not fully characterized. This study assessed fibrous proteins and cells in CVF of cadaveric livers. A total of 72 fibrotic veins out of 315 veins from 63 livers were studied. The diameters of the veins ranged from 25.4 to 123.7 μm (mean, 75.5 ± 18.3 μm). CVF was considered present when the rim thickness exceeded 9 μm (determined on Sirius red stained sections) coincident with fibrous tissue extension into the perivenous parenchyma. Immunoperoxidase staining revealed that the CV endothelium was positive for factor VIII‐related antigen, a vascular endothelial phenotypic marker. It also stained for CD31 and D34. Staining of basement membrane collagen IV was continuous around the CV endothelium, while laminin, another principal basement membrane protein, was not detected in the CV matrix. Thus, the CV appeared to lack a continuous endothelial basement membrane and a diffusion barrier between the CV circulation and perivenous parenchyma. Fibrillar collagens I, III and V and filamentous collagen VI were robust in the rim of the fibrotic vein. The codistribution of these collagens rendered thickening of the CV as well as provided strength to the CV matrix scaffold. Elastic fibers were noted occasionally among collagen bundles. Myofibroblasts, identified by a‐smooth muscle actin, were the dominant fibrogenic cells producing the fibrous matrix. Few mononuclear cells were also present. The deposition of collagenous fibrous tissue in the rim could occlude the CV endothelial openings, reducing the efficiency and increasing the resistance of sinusoidal blood flow into the CV. This may cause sinusoidal hypertension, impair hepatocyte functions and enhance disease susceptibility of the aged people.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Chemopreventive effect of capsaicin in experimentally induced hamster buccal pouch carcinogenesis (Immunohistochemical study Bcl-2)

Aim:To investigate immunohistochemically the effect of chemopreventive capsaicin in epithelial alteration (Epithelial dysplasia) & carcinogenesis which induced in hamsters. Methods: Hematoxylin and Eosin (H&E) and Streptavidin-biotin immunoperoxidase staining techniques was used to detect the expression of Bcl-2 in three groups (G1, G2 and G3). Evaluation of immunostaining was done using computer image analyzer system & statistical analysis. Results: showed that there was a statistically highly significant difference (P G1, G3) (G3>G1). Conclusion:Capsaicin play an important role in reduction and prevention of transformation of epithelial dysplasia into malignant tumor especially oral squamous cell carcinoma. The liquid form of capsaicin may be considered as a new line of treatment in oral dysplasia or even in oral squamous cell carcinoma and the Bcl-2 expression evidenced this reduction of abnormality in epithelial dysplasia.

Subcutaneous panniculitis-like T-Cell lymphoma: A mixed diagnostic approach to diagnosing a vague clinical picture

Subcutaneous panniculitis-like T-Cell lymphoma (SPTCL) is a rare subtype of cutaneous T-cell lymphoma, and it has been associated with a range of clinical symptoms from mild to severe. Most commonly, this disease is described as following a slowly progressing course, associated with vague constitutional symptoms and good prognosis. This case report describes the clinical presentation and findings of SPTCL in a 31 year old female and describes the challenges of recognizing and properly diagnosing this disease. SPTCL has been described as a mimicker of other, more common and nonmalignant diseases of the skin, such as lupus panniculitis. This report highlights a variety of specific tests including immunohistochemical and immunoperoxidase staining, as well as genotypic analysis of T-cell receptors, that were effective in combination in isolating this diagnosis. Moreover, choice of treatment for these patients can be challenging, as an array of interventions have been described in past cases to treat SPTCL. This report recognizes the efficacy of a treatment course that included a six-cycle course of combined chemotherapy (vincristine, doxurubicine, cyclophosphamide, and prednisone, also known as CHOP) followed by weekly methotrexate and PET scan surveillance for two years. With both initial and maintenance therapy, this patient showed excellence response evidenced by a progressive decrease in metabolic activity of malignant lesions, lack of new lesions, and remaining without symptoms. While this disease is rare, it is important to include SPTCL in the differential when considering patients with a panniculitic picture.

FXR1 expression domain in Wilms tumor

Background/PurposeWilms tumor (WT) is the most common childhood kidney cancer globally. Our prior unbiased proteomic screen of WT disparities revealed increased expression of Fragile X-Related 1 (FXR1) in Kenyan specimens where survival is dismal. FXR1 is an RNA-binding protein that associates with poor outcomes in multiple adult cancers. The aim of this study therefore was to validate and characterize the FXR1 expression domain in WT. MethodsQuantitative FXR1 gene expression was compared between WT, adjacent, adult, and fetal kidney specimens. The cellular and subcellular expression domain of FXR1 was characterized across these tissues using immunoperoxidase staining. RNA-sequencing of FXR1 was performed from WT and other pediatric malignancies to examine its broader target potential. ResultsFXR1 was detected in all clinical WT specimens evaluated (n = 82), and as a result appeared independent of demographic, histology, or adverse event. Specific cytosolic staining was strongest in blastema, intermediate and variable in epithelia, and weakest in stroma. When present, areas of skeletal muscle differentiation stained strongly for FXR1. qPCR revealed increased FXR1 expression in WT compared to adult and adjacent kidney (p < 0.0002) but was similar to fetal kidney (p = 0.648). RNA-sequencing revealed expression of FXR1 in multiple pediatric tumors, greatest in rhabdomyosarcoma and WT. ConclusionsFXR1 was expressed consistently across this broad sampling of WT and most robustly in the primitive blastema. Notably, FXR1 labeled a specific self-renewing progenitor population of the fetal kidney.

Lymphatic vessels identified in failed corneal transplants with neovascularisation

BackgroundCorneal transplant failure with neovascularisation is a leading indication for full-thickness grafts in patients. Lymphangiogenesis is implicated in the pathology of graft failure, and here we systematically evaluate failed human...

Velogenic newcastle disease virus tissue tropism and pathogenesis of infection in chickens by application of in situ PCR, immunoperoxase staining and HE staining

Limited deep studies are available in the field of early stages of pathogenesis of Newcastle disease virus (NDV) infection and tissue tropism of NDV. In this study, 24 specific pathogen free (SPF) chickens of white leghorn breed were infected with Newcastle disease (ND) by intranasal administration of 10⁵ 50% EID50/0.1 mL of velogenic NDV (vNDV). A second group of 15 chickens were kept as a control group. Chickens were monitored every day to record clinical signs. Infected chickens were euthanized by cervical dislocation at successive times, namely at hours (hrs) 2, 4, 6, 12, days 1, 2, 4, and 6 post-inoculation (pi). Whereas, control group chickens were euthanized on days 0, 1, 2, 4, and 6 pi. Tissues of brain, trachea, lung, caecal tonsil, liver, kidney, spleen, heart, proventriculus, intestine, and thymus were collected, fixed in 10% buffered formalin, embedded in paraffin, and sectioned. HS staining, immunoperoxidase staining (IPS) and in situ PCR were applied. It was concluded that at hr 2 pi, virus seemed to be inclined to trachea and respiratory tract. Meanwhile, it attacked caecal tonsils, intestine and bursa of Fabricus. While primary viraemia was ongoing, virus created footing in kidney and thymus. At hr 4 pi, proventriculus, liver, and spleen were attacked. However, at hr 6 pi, brain and heart were involved. Secondary viraemia probably started as early as hr 12 pi since all collected tissues were positive. Tissue tropism was determined in trachea, caecal tonsil, liver, bursa of Fabricius, intestine, proventriculus, lung, spleen, thymus, kidney, heart, and brain.

Infectious bursal disease virus tissue tropism and pathogenesis of the infection in chickens by application of in situ PCR, immunoperoxase and HE staining

Infectious bursal disease is one of an OIE list of notifiable diseases. Chicken is the only host that manifests clinical signs and its pathogenicity is correlated with the distribution of antigens in organs. This study was conducted to determine disease pathogenesis and virus tissue tropism by in situ PCR, immunoperoxidase staining (IPS), and HE staining. Twenty four chickens were infected with very virulent Infectious Bursal Disease Virus (vvIBDV). Fifteen chickens were kept as a control group. Infected chickens were sacrificed at hrs 2, 4, 6, 12, days 1, 2, 4, and 6 post-inoculation (pi). While, control chickens were euthanized on days 0, 1, 2, 4, and 6 pi. Different tissues were collected, fixed in 10% buffered formalin, and processed. At hr 2 pi, virus was detected in intestinal, junction of the proventriculus and gizzard, cecal tonsil, liver, kidney, and bursa of Fabricius. At hr 4 pi, virus reached spleen, and at hr 6 pi, it entered thymus. At hr 12 pi, virus concentration increased in positive tissues. The latest invaded tissue was muscle on day 1 pi. Secondary viraemia occurred during 12–24 h pi. In situ PCR was the most sensitive technique to highlight obscure points of infection in this study.

A double dose of pseudolymphoma: Cutaneous intravascular CD30+ pseudolymphoma arising in acral pseudolymphomatous angiokeratoma – a case report

Cutaneous intravascular CD30+ pseudolymphoma is a rare entity with only a handful of cases reported thus far in the literature. This case study reports a patient who presented with a non-healing wound of the forearm, which on excision biopsy was found to have the histological features of an acral pseudolymphomatous angiokeratoma, also known as papular angiolymphoid hyperplasia. Distended lymphatics within the lesion were filled with a monotonous population of intermediate sized lymphocytes with distinct nucleoli and considerable associated apoptotic debris. Immunoperoxidase staining showed almost uniform staining for CD4 and around 50% staining for CD30. The occurrence of cutaneous intravascular CD30+ pseudolymphoma has previously been reported in the setting of a regressing keratoacanthoma of the cheek1 and has been associated with chronic inflammation after trauma.2 In this case the finding was interpreted as a florid reactive hyperplasia, with the recommendation for follow up and T-cell clonality studies should recurrence occur. To our knowledge, no recurrence has occurred 20 months post initial presentation. Awareness of this benign cutaneous lymphoproliferation as a reactive process is necessary in distinguishing it from malignant intravascular T-cell lymphomas.