Roland Schwarting

Thymidine Phosphorylase Expression Is Associated With Response to Capecitabine Plus Irinotecan in Patients With Metastatic Colorectal Cancer

PURPOSEnTo evaluate the clinical activity and toxicity of capecitabine plus irinotecan as first-line therapy for patients with metastatic colorectal cancer (mCRC), and to describe the association of expression of thymidine phosphorylase (TP), thymidylate synthase (TS), and dihydropyrimidine dehydrogenase (DPD) with antitumor activity.nnnPATIENTS AND METHODSnPatients with previously untreated mCRC received irinotecan days 1 and 8 intravenously, and capecitabine days 2 to 15 orally in 21-day cycles. Doses were irinotecan 125 mg/m2 and capecitabine 1,000 mg/m2 bid (n = 15; cohort 1), or irinotecan 100 mg/m2 and capecitabine 900 mg/m2 bid (n = 52; cohort 2). Tissues from primary and metastatic sites were assessed for TP, TS, and DPD gene and protein expression.nnnRESULTSnAn unacceptable level of GI toxicity in the first 15 patients led to a protocol modification in starting doses. The response rate was 45% (30 of 67 patients). Overall survival was associated with TP expression assessed by immunohistochemistry in both primary tumors (P = .045) and metastases (P = .001). Objective tumor response was associated with TP expression in primary tumors (odds ratio, 4.77; 95% CI, 1.25 to 18.18), with a similar trend in metastases (odds ratio, 8.67; 95% CI, 0.95 to 79.1). TP gene expression in primary tumors was also associated with response.nnnCONCLUSIONnThese data indicate that capecitabine plus irinotecan is an active regimen against mCRC. The biomarker analysis (including metastatic tissue) was feasible in a multicenter setting, and provides preliminary evidence that TP expression may be a predictive marker for response.

Mucosa specific lymphocytes in the human conjunctiva, corneoscleral limbus and lacrimal gland

Conjunctiva associated lymphoid tissue shows several similarities to mucosa associated lymphoid tissue of the gut and respiratory tract. These similarities have been described in relation to lymphocyte subpopulations and epithelial cell morphology. However, unlike the lymphoid tissue of the gut and respiratory tract, mucosa specific lymphocytes have not been described in the ocular mucosa. In this study we demonstrated the presence of mucosa specific lymphocytes bearing the Human Mucosal Lymphocyte-1 antigen (beta 7 integrin), in the human conjunctiva, limbus and lacrimal gland. The distribution of this subset of lymphocytes corresponded to the distribution of CD8+ T-cells and was found maximally in the epithelium of the epibulbar conjunctiva and in the lacrimal gland. The Human mucosal lymphocyte antigen may function to determine mucosal homing of this particular subset of CD8+ T-cells, which in turn, may have special function in immunological defense and tolerance mechanisms occurring at mucosal surfaces.

An evaluation of the performance of sysmex XE-2100 in enumerating nucleated red cells in peripheral blood

CONTEXTnAutomated methods of enumerating nucleated red blood cells (NRBCs) in blood are gaining acceptance in many laboratories.nnnOBJECTIVEnTo evaluate the performance of Sysmex XE-2100 in enumerating NRBCs in peripheral blood.nnnDESIGNnAutomated relative number of NRBCs per 100 white blood cells (NRBC%) results for a total of 460 specimens run on the XE-2100 were compared with manual NRBC% results obtained by performing a 100-cell differential on a Wright-stained smear prepared from each specimen. To assess within-day reproducibility, 64 specimens were rerun on the XE-2100, and a second 100-cell differential was performed on the original blood smear. Excel software was used for data analysis.nnnRESULTSnRegression analysis of automated NRBC% versus manual NRBC% yielded a correlation coefficient of 0.9712. No NRBCs were seen in the blood smear on 35 (15.1%) of 232 specimens with automated NRBC% of 0.1 to 1.9. The XE-2100 generated an NRBC% of 0.0 on 5 (6.8%) of 74 specimens, revealing 1 NRBC per 100 or more white blood cells by blood smear examination. The mean percent difference between duplicate automated results was 16.7 compared with 78.1 for the duplicate manual results. There were 9 instances in which the XE-2100 either did not detect the presence of more than 8 NRBCs per 100 white blood cells or generated an automated NRBC% of 18.1 or 18.8 when the smear revealed none. All of these were, however, flagged for smear review.nnnCONCLUSIONSnOverall correlation between the automated and manual results was excellent. The automated method revealed better precision than the manual method. The number of specimens with false automated results was very small, and all were flagged for verification by a smear review.

Effects of Storage of Blood at Room Temperature on Hematologic Parameters Measured on Sysmex XE-2100

Blood specimens are frequently delivered to the clinical laboratory after a significant postcollection interval. On weekends, this interval may exceed 72 hours. When such a specimen arrives, the laboratory must decide whether to accept or reject the specimen. If accepted, the laboratory must then decide whether to perform all of the tests ordered or only those deemed appropriate given the age of the specimen, and what comments should be appended to the reported results regarding the reliability or limitations of the analysis. Storage of K2-EDTA-anticogulated blood at room temperature for up to 4 days caused changes in some but not all hematologic parameters measured on Sysmex XE-2100 analyzer. Specifically, the WBC, RBC, HGB, and PLT were found to be stable for up to 3 days. Retic % and #, but not the IRF, were stable for the duration of the study. Diff results may be unreliable or voted out by the analyzer on specimens that are 2 day old or older. The NRBC counts changed little on the average but revealed consistently wide 95% confidence intervals throughout.

Renal cell carcinoma in the presence ofadult polycystic kidney disease

Abstract A case of autosomal dominant polycystic kidney disease associated withwidely metastatic renal cell carcinoma is reported. The patient had presented with pneumothorax, weight loss, leukocytosis, lytic bone lesions, and hypercalcemia. Despite intensive diagnostic search for a neoplasm, no firm evidence of malignancy was found. However, at the autopsy, widely metastatic, papillary renal cell carcinoma was found originating in the left kidney. Many metastases showed central necrosis mimicking small cysts.

Analysis of Clonality Using X-Linked Polymorphisms in a Patient With Multiple Myeloma and Myelofibrosis

We describe a patient who presented with a neutrophilic leukocytosis, normal karyotype, and IgAλ multiple myeloma. One year after diagnosis she developed diffuse myelofibrosis as well as multiple lytic lesions of bone. Given the myeloproliferative features of her case, the clonality of her peripheral leukocytes was determined prior to treatment. Analysis of X‐chromosome inactivation at the X‐linked human androgen‐receptor gene locus (HUMARA) proved that granulopoiesis was polyclonal. Subsequent treatment of the myeloma reversed with myelofibrosis and normalized her WBC count. This is the first case of multiple myeloma with myelofibrosis in which a concomitant clonal myeloproliferative disease was ruled out at a genetic level. The myeloproliferative features in this case are presumed to be induced by cytokines produced by the plasma cell clone. Am. J. Hematol. 59:79–82, 1998.

An Assessment of the Coulter Gen•S Automated Flagging System

instrumentation [generalist | hematology | cytology] An Assessment of the Coulter Gen•S Automated Flagging System Gene L. Gulati, PhD,1,2 William Kocher, MD,1,2 Roland Schwarting, MD,1,2 Lawrence J. Hyland, MD,1,2 Ali Issa, BSMT,1 Robert Arwood, BSMT,1 and Manmohan Dhanjal, BSMT(ASCP)SH1 From the 1Hematology Laboratory, Thomas Jefferson University Hospital, and the 2Department of Pathology, Jefferson Medical College, Philadelphia, PA

Identification of an S-antigen-like molecule in human choroid plexus and cerebrospinal fluid

Sensitisation to retinal S-antigen has been implicated in the pathogenesis of several clinical forms of posterior uveitis. S-antigen-like molecules have recently been demonstrated in the brain and choroid plexus of experimental animals. We used a panel of four monoclonal antibodies (MAbs), MAbF4-C1, MAbC10-C10, MAbA2-G5 and MAbA9-C6, which define specific epitopes in the amino, mid and carboxyl terminal portions of S-antigen in order to identify an S-antigen-like molecule in human choroid plexus and cerebrospinal fluid (CSF). Three MAbs, MAbF4-C1, MAbC10-C10 and MAbA9-C6, localised an S-antigen-like molecule to the cytoplasm of the epithelial cells of the human choroid plexus. Polymerase chain reaction of cDNA from choroid plexus verified the presence of S-antigen homologues in the choroid plexus. The presence of an S-antigen-like molecule in the CSF was demonstrated by western blots in seven CSF samples from patients with a variety of neuropathological disorders. It is proposed that immunological cross-reactivity and biochemical similarity between retinal S-antigen and an S-antigen-like molecule in human choroid plexus and CSF could form a basis for neurological manifestations observed in certain clinical forms of uveitides.

Recognition of normal, neoplastic, and fetal airway epithelial cell membranes by two monoclonal antibodies

The reactivity of two rat monoclonal antibodies was studied. These antibodies, A2R and A2C, bind a 32 kDa alveolar type II cell membrane receptor for surfactant protein A. A2R and A2C also bind apical cell membranes of ciliated and nonciliated cells of the conducting airways. Because this reactivity suggested possible utility in targeting those cells for therapeutic gene transfer, the binding activity of these two antibodies was examined in human tissues. In conducting airways, A2R and A2C bound apical epithelial cell membranes throughout the embryologic period studied: from 15 weeks of gestation, through maturity. Reactivity was more restricted to ciliated cells of the airways as maturation progressed. In the peripheral lung, A2C and A2R only bound most cells in the early developing lung, but mainly type II cells in mature lungs. Other normal tissues recognized by these antibodies included crypt lining cells of the adult and fetal stomach, large bile duct epithelium, and pancreatic acinar cells. All of these cells derive from embryonic foregut endoderm. Other normal tissues, both of endodermal and nonendodermal origin, were negative. Pulmonary carcinomas were studied. A2C and A2R recognized all non-small cell carcinomas of the lung tested. In contrast, none of the small cell carcinomas or carcinoid tumors of the lung were recognized by these antibodies. The function of p32 in these diverse cell types is not clear, but whatever its role in these tissues, antibodies versus p32 may potentially be used to target gene or drug therapy to the normal or malignant cells they recognize.

Extramedullary myeloid cell tumor of the urinary bladder in a patient with myelodysplastic syndrome.

We report a case of extramedullary myeloid cell tumor of the urinary bladder in an elderly male with a three year history of myelodysplastic syndrome (refractory anemia with excess blasts), noninvasive papillary transitional cell carcinoma of the urinary bladder, and in situ transitional cell carcinoma of the left ureter. Light microscopy demonstrated a poorly differentiated neoplasm composed of medium to large cells with eosinophilic cytoplasm. The tumor cells showed immunohistochemical expression of myeloperoxidase, lysozyme, CD15, CD68 and CD43. Bone marrow examination following cystectomy demonstrated refractory anemia with excess blasts (6-10%) and a normal karyotype. Cytogenetics, approximately 1 year after cystectomy, demonstrated a deletion of the short arm of chromosome number 12. Four years after presentation, the patient succumbed to pulmonary aspergillosis.