Strong Annexin A10 Expression Supports a Pancreatic Primary and Combined Annexin A10, Claudin 18, and SOX2 Expression Supports an Esophagogastric Origin in Carcinomas of Unknown Primary.

Primary tumor site determination for gastrointestinal (GI) tract and pancreaticobiliary (PB) tree carcinomas that present as metastasis of unknown primary can be problematic. Annexin A10 (ANXA10), claudin 18 (CLDN18), and trefoil factor 1 (TFF1) have been identified through expression profiling as markers of gastric lineage commitment; sex-determining region Y (SRY)-box transcription factor 2 (SOX2) expression has been reported in several tumor types, including gastric adenocarcinomas. We evaluated the diagnostic utility of immunohistochemistry for ANXA10, CLDN18, SOX2, and TFF1 for determining the site of origin for GI/PB adenocarcinomas. Immunohistochemistry for all 4 markers was performed on tissue microarrays including 559 GI/PB tumors and 421 other tumors. H-scores were calculated as the product of the intensity (0 to 3) and extent (percentage, 0% to 100%) of staining. Positive staining was defined as >5% staining. ANXA10 expression was most frequent in pancreatic adenocarcinomas when compared with all other GI/PB tumors (96.4% vs. 43.5%, P <0.001). Strong staining for ANXA10 (H-score ≥200) distinguished pancreatic ductal adenocarcinoma from intrahepatic cholangiocarcinoma and adenocarcinomas of the gallbladder and colorectum (69.6% vs. 0%, P <0.001). Triple positivity for ANXA10, CLDN18, and SOX2 was more frequent in esophagogastric tumors than in other GI/PB tumors (22.6% vs. 4.1%; P <0.001). TFF1 expression was observed in nearly all tumor types. Staining for ANXA10, CLDN18, and SOX2 as part of a panel may aid in distinguishing esophagogastric adenocarcinomas from lower GI/PB tumors. ANXA10 staining may be particularly useful in distinguishing pancreatic adenocarcinomas from intrahepatic cholangiocarcinoma and adenocarcinomas of the gallbladder and colorectum.

False positive testing for sickle hemoglobin in a blood donor with mild erythrocytosis and hemoglobin Geldrop St. Anna.

Solubility testing for sickle hemoglobin is commonly performed to identify blood suitable for patients with sickle cell disease. A 32-year-old Caucasian male blood donor's unit screened positive for sickle hemoglobin via solublity testing (Streck). As the donor was considered low risk for being positive for hemoglobin S (HbS), he self-referred to hematology for further evaluation. Testing with hemoglobin electrophoresis revealed the patient to be negative for HbS; however, 42 % fetal hemoglobin (HbF) was noted. Since this was higher than typically seen in hereditary persistence of HbF, deoxyribonucleic acid (DNA) sequencing of hemoglobin (Hb) was ordered through a referral laboratory. Hb gene sequencing revealed the patient to be heterozygous for Hb Geldrop St. Anna, a rare Hb variant. This variant has previously been shown to migrate in the HbF region with alkaline electrophoresis. The workup demonstrated that the oxygen dissociation curve was left-shifted consistent with slightly increased oxygen affinity of this variant. The patient's hematocrits (Hct) from his past donations were 53 % and 54 % about two years apart and his Hct at his hematology evaluation was 53 %. This report describes the first case of Hb Geldrop St. Anna in the United States and was associated with a false positive HbS screen. This Hb variant is considered to be benign and has an increased oxygen affinity that is associated with mild erythrocytosis. The donor was allowed to continue donating blood products.

Nicotinamide phosphoribosyltransferase expression and clinical outcome of resected stage I/II pancreatic ductal adenocarcinoma.

BACKGROUND: Nicotinamide phosphoribosyltransferase (NAMPT) plays a key role in the biosynthesis of nicotinamide adenine dinucleotide (NAD+), which is a vital cofactor in redox reactions and a substrate for NAD+ consuming enzymes including CD38, PARPs and sirtuins. NAMPT over-expression has been shown in various cancers and its inhibition decreases cancer cell growth, making it an attractive therapeutic target. Here we examine the NAMPT expression in a large cohort of resected stage I/II pancreatic ductal adenocarcinomas (PDAs) and correlate its expression with clinical outcomes and pathologic features. METHODS: A retrospective review of patients with PDAs was conducted at a single institution. Tissue microarrays (TMAs) containing primary PDAs and their metastatic lymph nodes (mLNs) were constructed and stained for NAMPT expression. Each TMA core was evaluated for staining intensity of cancer cells (0 = no staining, 1+ = weak, 2+ = moderate, 3+ = strong) and a mean score was calculated for each case with at least two evaluable cores. NAMPT expression was correlated with clinicopathological variables using chi-squared or Fisher's exact test, and t-tests for categorical and continuous variables, respectively. Survival probabilities were estimated and plotted using the Kaplan-Meier method. Cox proportional hazards regression was used to assess the effects of NAMPT staining values on recurrence-free survival (RFS) and overall survival (OS). This study was conducted under an approved IRB protocol. RESULTS: 173 primary PDAs had at least 2 TMA cores with identifiable cancer cells. The mean IHC score was 0.55 (range: 0 to 2.33). The mean IHC score of mLNs was 0.39 (range: 0-2), which was not significantly different from their primary tumors (mean IHC score = 0.47, P = 0.38). Sixty-four percent (111/173) of PDAs were positive for NAMPT staining. Stage II tumors were more likely to be positive (68% of 151 vs 41% of 22; P = 0.01). Non-obese non-diabetic patients were more likely to have NAMPT+ tumors (43.7% vs. 27.9%, P = 0.04). While RFS and OS were not statistically different between NAMPT+ vs. NAMPT- PDAs, patients with NAMPT- tumors tended to have a longer median OS (26.0 vs. 20.4 months, P = 0.34). CONCLUSION: NAMPT expression was detected in 64% of stage I/II PDAs and up to 72% in non-obese non-diabetic patients. Frequency of NAMPT expression correlated with pathological stage, consistent with published literature regarding its role in cancer progression. While RFS and OS were not statistically significantly different, patients with NAMPT+ PDAs tended to have a shorter survival. Thus, NAMPT inhibition may prove beneficial in clinical trials.

Traditional versus reverse syphilis algorithms: A comparison at a large academic medical center.

OBJECTIVES: An increasing number of institutions are transitioning from the traditional syphilis testing algorithm (initial screening with nontreponemal tests) to the 'reverse' algorithm (initial screening with treponemal tests such as syphilis IgG). The aim of this study was to evaluate the switch in syphilis algorithm at an academic medical center with a population with low syphilis prevalence. DESIGN AND METHODS: We performed a six-year retrospective study at the University of Iowa Hospitals and Clinics, an academic medical center, comparing the traditional algorithm (n=12,612) with the reverse algorithm (n=10,453). False positives were considered to be positive screens with negative confirmatory testing. RESULTS: Using the traditional algorithm, 93 samples (0.7% of total) screened positive with RPR, with 40 of these samples having negative TP-PA testing (43% of positive screens, 0.3% of total). Using the reverse algorithm, 110 screened positive with syphilis IgG (1.1% of total), and 33 of these samples had both negative RPR and TP-PA (30% of positive screens, 0.3% of total). In both algorithms, higher RPR titers and syphilis IgG values were associated with increased probability of positive confirmation. CONCLUSIONS: In this study at an academic medical center, the reverse algorithm had significantly more total positive screens than the traditional algorithm. Both algorithms produced equivalent rates of active infection. The quantitative difference in positives between the two algorithms are the category of patients who are syphilis IgG positive, RPR non-reactive, and TP-PA reactive. Specimens with higher RPR titers and syphilis IgG values are more likely to confirm positive.

Buffer Standards for the Physiological pH of the Zwitterionic Compound, DIPSO from 5 to 55 degrees C.

The values of the second dissociation constant, pK(2), and related thermodynamic quantities of 3-[N,N-bis (2-hydroxyethyl)amino]-2-hydroxypropanesulfonic acid (DIPSO) have already been reported over the temperature range 5 to 55 degrees C including 37 degrees C. This paper reports the pH values of four NaCl-free buffer solutions and four buffer composition containing NaCl salt at I = 0.16 mol.kg(-1). Conventional pa(H) values are reported for all eight buffer solutions. The operational pH values have been calculated for four buffer solutions recommended as pH standards, at 25 and 37 degrees C after correcting the liquid junction potentials with the flowing junction cell.

Buffer Standards for pH Measurement of N-(2-Hydroxyethyl)piperazine-N'-2-ethanesulfonic Acid (HEPES) for I = 0.16 mol.kg from 5 to 55 degrees C.

The values of the second dissociation constant, pK(2) of N-(2-hydroxyethyl) piperazine-N'-2-ethanesulfonic acid (HEPES) have been reported at 12 temperatures over the temperature range 5 to 55 degrees C, including 37 degrees C. This paper reports the results for the pa(H) of eight isotonic saline buffer solutions with an I = 0.16 mol*kg(-1) including compositions: (a) HEPES (0.01 mol*kg(-1)) + NaHEPES (0.01 mol*kg(-1)) + NaCl (0.15 mol*kg(-1)); (b) HEPES (0.02 mol*kg(-1)) + NaHEPES (0.02 mol*kg(-1)) + NaCl (0.14 mol*kg(-1)); (c) HEPES (0.03 mol*kg(-1)) + NaHEPES (0.03 mol*kg(-1)) + NaCl (0.13 mol*kg(-1)); (d) HEPES (0.04 mol*kg(-1)) + NaHEPES (0.04 mol*kg(-1)) + NaCl (0.12 mol*kg(-1)); (e) HEPES (0.05 mol*kg(-1)) + NaHEPES (0.05 mol*kg(-1)) + NaCl (0.11 mol*kg(-1)); (f) HEPES (0.06 mol*kg(-1)) + NaHEPES (0.06 mol*kg(-1)) + NaCl (0.10 mol*kg(-1)); (g) HEPES (0.07 mol*kg(-1)) + NaHEPES (0.07 mol*kg(-1)) + NaCl (0.09 mol*kg(-1)); and (h) HEPES (0.08 mol*kg(-1)) + NaHEPES (0.08 mol*kg(-1)) + NaCl (0.08 mol*kg(-1)). Conventional pa(H) values, for all eight buffer solutions from 5 to 55 degrees C have been calculated. The operational pH values with liquid junction corrections, at 25 and 37 degrees C have been determined based on the NBS/NIST standard between the physiological phosphate standard and four buffer solutions. These are recommended as pH standards for physiological fluids in the range of pH 7.3 to 7.5 at I = 0.16 mol*kg(-1).