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Background & Objectives: Recurrent Aphthous Stomatitis (RAS) is one of the most common chronic ulcerative lesions of the oral mucosa and its development may be associated with oxidative stress. The aim of this study was to evaluate salivary Total Antioxidant Capacity (TAC) in patients with minor RAS. Materials & Methods: In this case-control study, 27 patients with minor RAS and 28 age- and sex-matched controls without RAS were enrolled. TAC was measured in unstimulated saliva for patients (during active lesion phase and after healing) and controls by immunologic assay. Data were analyzed by SPSS 18 using paired and unpaired t-tests (P Results: Salivary TAC levels of patients presented a significant increase from active lesion phase (0.26±0.16) to healing time (0.43±0.41); (P=0.034). There was no significant difference in the level of salivary TAC between patients during active lesion phase and controls (0.24±0.13); (P=0.641). Conclusion: Increasing level of salivary TAC may be involved in remission of RAS lesions, suggesting its evaluation in future studies.

Low folate concentrations are inversely associated with birth defects, including neural tube defects, congenital heart disease and oral clefts. Conversely, high folate concentrations may be associated with adverse outcomes, including increased risk of colorectal cancer among those with pre-existing neoplasms. The purpose of our study was to investigate the folate status of a nationally representative sample of Canadians, including a subset of women of childbearing age.We examined red blood cell folate concentrations among members of the general population aged 6-79 years (n = 5248) and separately among women of childbearing age (15-45 yr, n = 1162), as recorded by the Canadian Health Measures Survey and measured by immunologic assay. We assessed the data for significant differences by age, sex and socioeconomic status.Less than 1% of Canadians showed folate deficiency (red blood cell folate305 nmol/L) and 40% showed high folate concentrations (1360 nmol/L). Among women of childbearing age, 22% showed concentrations below those considered optimal for maximal neural tube defect-risk reduction (906 nmol/L). Significant differences by age and socio-economic status, but not sex, were evident in median red blood cell folate concentrations, although concentrations in all groups exceeded recommended levels. No differences by age or income were found among women of child-bearing age.Folate deficiency is virtually nonexistent in the Canadian population, although high folate concentrations are evident. Additional research is needed to better understand the determinants of red blood cell folate among women of childbearing age who have concentrations below levels that are maximally protective against neural tube defects. Ongoing monitoring of the folate status of Canadians and the relationship between red blood cell folate and health outcomes is warranted.

29th Annual Jackson Hole, Wyoming, Update in Clinical Immunology, Microbiology and Infectious DiseasesSalt Lake city, UT, USA, 9–13 July 2012The 29th Annual Jackson Hole, Wyoming, Update in Clinical Immunology, Microbiology and Infectious Diseases was held from 9 to 13 July 2012. This postgraduate, continuing medical education course of the University of Utah’s Department of Pathology (UT, USA) is designed for laboratorians, clinical pathologists, pathologists, clinicians, clinical immunology and infectious disease specialists and medical technologists, as well as residents and fellows training in immunology, microbiology or infectious diseases.

Companion diagnostics, tests that purport to classify patients into “responders” and “non-responders” for a specified targeted therapy, demand methods that quantify the actual amount of the corresponding target molecule in tumors from these patients. Various methods are employed depending upon the nature of the target. Many of the candidate therapeutic agents target the abnormal expression of a protein, the detection of which lends itself to an immunohistochemistry (IHC) approach. This review focuses on IHC with formalin-fixed paraffin-embedded (FFPE) tissues for purely pragmatic reasons; first, the morphologic information pertaining to the tumor is of value and should not be discarded as in extraction type assays; second, FFPE tissues are mostly what we have to hand at the time that the diagnostic question is posed. During the four decades of employment of IHC involving the production of a variety of special stains used in the diagnosis or classification of tumors, we have acquired some bad habits, essentially when judging the IHC result via the perception of a “good” stain that “pleases the eye” of the user pathologist, and nothing more. This review takes, as its basic premise, the notion that IHC can be upgraded from its use as a qualitative special staining method to an accurate and reliable quantitative “tissue-based immunologic assay”. If accomplished, this enhanced IHC assay would serve accurately to quantify proteins in tissue sections, analogous to the use of the ELISA (enzyme-linked immunosorbent assay) method in the clinical laboratory. The necessary steps for converting IHC to a tissue-based ELISA-like immunoassay of immediate practical use are reviewed with constructive suggestions for steps that can be (must be) taken to achieve the practical reality of quantitative in situ proteomics.

We sought to determine if serum total iron-binding capacity (TIBC) is equivalent to serum transferrin (TRF) so that a low-cost colorimetric chemical assay for unsaturated iron-binding capacity (UIBC) could be substituted for a high-cost immunologic assay for TRF. Our study design included independent and blinded measurements of UIBC, serum iron, and TRF concentrations in human serum samples. Data from five independent correlation studies carried out at three different Quest Diagnostics laboratories were combined into one data set containing 570 paired results for TIBC and TRF. r2 was 0.941 when three outliers were eliminated from the 570-sample data set. Scatter about the regression line was fully accounted for by the CVs for the TIBC and TRF assays. When each test is measured precisely and without bias, the ratio of TIBC (μmol/L) to TRF (g/L) in SI units is close to the theoretically expected value of 25.0.

SUMMARY The discipline and techniques of immunology have been an important part of recent advances in genitourinary oncology. Although the initial promise of tumor immunology has not been realized in man, discoveries in the laboratory are revitalizing the expectation that this field will yet be of significant assistance in the diagnosis and treat ment of cancer. Already tumor immunology studies along with new immunologic assay techniques have produced clinically important tumor markers, such as alpha-fetoprotein, human chorionic gonadotropin, and prostatic acid phosphatase, and have intensified the search for other markers. Finally, research activity is increasingly focusing on the tumor cell itself, again often using immunologic methods. Although most of this activity is still experimental, analysis of bloodgroup antigens in transitional cell carcinoma has produced sufficient data to suggest that such an approach will soon be a part of the urologist’s clinical armamentarium in his efforts to treat patients with bladder tumors.

SummaryProtein C is a vitamin K dependent protein involved in blood coagulation. A congenital deficiency in protein C antigen - which inherits as an autosomal dominant disorder - has been reported to be associated with a high risk for thrombo-embolic disease at relatively young age. In the present paper we report on the development of a functional assay for plasma protein C. In this assay protein C is adsorbed to Al(OH)3, eluted and activated by thrombin, after which the concentration of the activated protein C is measured with a peptide substrate (S2366). Normal values for protein C activity and protein C antigen were determined in healthy volunteers and patients on stable oral anticoagulant treatment. Protein C activity and antigen levels were compared in 28 patients from 9 different pedigrees with both congenital protein C deficiency and thrombotic disease. Two types of protein C deficiency could be recognized: in type I the deficiency is due to the absence or reduced presence of protein C molecules, while in type II the deficiency is caused by the presence of an abnormal protein C molecule with strongly reduced functional activity.

A fast functional assay for protein C was evaluated and compared with a traditional functional and an enzyme linked immunosorbent assay in parallel for the same plasma samples derived from 43 healthy subjects, 12 patients with severe hepatic dysfunction, and 23 patients under stable oral anticoagulation. By all three test systems significantly lower levels of protein C were obtained in both groups of patients compared with normal subjects (p less than 0.0001). No significant between - assay differences were found in normal subjects and in patients with hepatic dysfunction; by correlation analysis coefficients higher than 0.8 were calculated between the measurements of the three tests. In patients under stable oral anticoagulation, however, the immunologic test yielded higher values than the traditional (p less than 0.05) and, more pronounced, the fast functional assay (p less than 0.0001); no or only borderline significant correlations between the results were found. In these patients protein C levels measured with the traditional functional assay were in the same range as the activity levels of factors II, VII, IX, and X, whereas the fast functional test yielded significantly lower levels. The presented results indicate that very similar protein C levels were obtained with both functional and the immunologic assay except in patients under oral anticoagulation.

This investigation was undertaken to determine the prevalence and degree of malnutrition among patients with chronic obstructive pulmonary disease and to determine if a relationship exists between nutritional assessment parameters and pulmonary function. Thirty-seven patients admitted for the treatment of chronic obstructive pulmonary disease to a respiratory care unit of a Veterans Administration facility were studied. Nutritional status was evaluated using anthropometric measurements, laboratory data and immunologic assay. Deficits in anthropometric measurements were frequent and severe in this group of patients. Furthermore, our data demonstrated that patients with a forced vital capacity

Current assay methods do not define the absolute concentrations of calcitonin in plasma of normal man. In our experience, the concentration was

An immunologic method of assay of HCG in serum has been developed using the hemagglutination inhibition reaction. Serum was treated with acetone and the HCG containing fraction was washed with absolute alcohol and ether. Immunologic assay of HCG in serum of 311 normal pregnant women revealed results comparable with those obtained by the various biological methods of assay both in the shape of the normal curve and in the actual concentration of the hormone measured in international units. In addition, the concentration of HCG in serum as compared with morning urine, both being assayed immunologically, has been found to be very similar. Finally, the rate of disappearance of the hormone from the serum as well as its presence in amniotic fluid in significant amounts has been determined.

A brief description of our experience with the immunoassay of insulin by the technic of Hales and Randle is given. Insulin values determined in twelve normal children and in seven cases of untreated diabetes mellitus in the fasting state and following a glucose load are presented. A newly diagnosed untreated child with diabetes mellitus had a low fasting insulin and gave very little response to test meal of glucose. Implications of these findings are discussed briefly.

Summary: Eigtheen couples (12.9%) were selected from a group of 138 infertile couples for a special study about the reliability of the SCMC test, when compared with spermanti-body titres in the cervical mucus and or seminal plasma and corticosteroides as treatment for immunologic infertility. It can be demonstrated that there exists a reliability of the SCMC test as an immunologic assay for sperm antibody activity in the semen and the cervical mucus. Zusammenfassung: Der SCMC-Test: Ein genauer Indikator fur Spermatozoen-Antikorper Aus einem Kollektiv von 138 infertilen Paaren wurden 18 Paare (12.9%) fur eine Studie ausgewahlt, in welcher die Zuverlassigkeit des SCMC-Tests im Ver-gleich zu den Spermatozoenantikorper-Titern im Cervikalschleim und/oder im Seminal-plasma und zu einer Corticosteroidtherapie zur Behandlung der immunologischen Inferti-litat untersucht wurde. Es kann gezeigt werden, das fur den SCMC-Test eine Zuverlassigkeit existent ist, sodas dieser als ein immunologischer Test fur die Spermatozoenantikor-per-Aktivitat im Sperma und im Cervicalschleim gelten kann.

A B S T R A C T A heat-stable humoral substance (coagulopoietin-X) is present in rabbits partially depleted of Factor X, which is capable of raising Factor X levels when injected into recipient rabbits. Rabbits were partially depleted of Factor X by slow infusion of a globulin fraction of goat anti-rabbit Factor X antibody. This resulted in the reduction of Factor X to 40-50% of nonnal at 1 h and 60-70% of normal at 6 h. No effect was noted on levels of Factors II, V, or VII. Plasma from these animals, when injected into 10 recipients, specifically raised Factor X levels when measured by four different assays: one-stage assay with bovine VII- and X-deficient plasma and Russell's viper venom; one-stage assay with human X-deficient plasma and thromboplastin; chromogenic substrate assay with Russell's viper venom; and an immunologic assay (Laurell technique). No rise was noted in two control experiments in which normal plasma was injected into recipient rabbits from 2 rabbits injected with a globulin fraction of normal goat serum, nor in 12 rabbits injected with plasma from normal rabbits, nor in 5 rabbits injected with boiled plasma from normal rabbits. The rise in biologic activity of 120-150% of base line was significantly greater than the rise in immunologic activity of114-117% ofbase line (P < 0.05) on 3 different days, suggesting the production of a molecule with greater specific activity rather than increased protein synthesis.

A kindred with a history of multiple thromboses was studied for coagulant abnormalities. A deficiency of plasma antithrombin III was found in approximately half of the 13 family members by either coagulant or immunologic assay. No clear relationship between plasma antithrombin III deficiency and a history of thrombosis was present. Platelet antithrombin assays were studied in the same family. Ten of the 13 members were deficient. None of the three who was normal had a history of thrombosis. The plasma antithrombin III assays could be influenced by heparin or coumarin treatment. The same medications were without consistent effect on platelet antithrombin. On the basis of these findings, it is proposed that familial hypercoagulability in some cases may be due to a platelet antithrombin deficiency and that the plasma antithrombin III deficiency is a secondary effect.

Quantitative immunologic assay of the plasma of the proposita in a family with hereditary prothrombin deficiency demonstrated the defect to be due to an actual deficiency of the prothrombi...

Assays of urinary human chorionic gonadotropin (HCG) in normal and abnormal pregnancies performed by a hemagglutination inhibition method are presented. Antisera were produced in rabbits. An HCG preparation of pooled urine of pregnant women was used as a laboratory standard. 1st morning urine samples were obtained from a large group of patients mostly in aid of diagnosis of pregnancy. Patients with a variety of pathologic conditions also supplied specimens. There were a total of 1021 assays on 691 patients. It was possible to follow only 492 patients for 588 assays. There were 10 false positive and 2 false negative results for an overall accuracy of 98.0%. The mean level of HCG in normal pregnancies peaked at about 70,000 IU in the first trimester then fell to a constant level of 20,000 IU during the 2nd half of the pregnancy. The 95% confidence limits of the levels in normal pregnancy were from 10,000 to 350,000 IUD in the latter part of the first trimester and 2500-150,000 in the remainder. The levels of HCG in patients with clinical evidence of threatened abortion were determined. There were 86 such pregnancies, 47 of which had spontaneous abortion, while the remaining 39 went to term. The mean levels of the urinary HCG in the patients that aborted were consistently lower than those with normal pregnancies. Only during the period of 57-110 days after the last menstrual period were the values significant (P less than .001). 4 out of 6 patients with hydatidiform moles had titers of HCG greater than the normal range. HCG tests are also useful in following patients treated with chemotherapy for choriocarcinoma. The immunologic assay of HCG is a rapid, uncomplicated method of determining the level of HCG. It is of value in aiding the clinician in diagnosis, prognosis, and management of certain pathologic conditions of pregnancy.

Plasma and urinary prothrombin conncentration and plasma prothrombin activity were measured in a group of 17 patients with the nephrotic syndrome. An immunologic assay using a monospecific antibody against human prothrombin was employed in the measurement of prothrombin concentration in the plasma and urine. Prothrombin-deficient plasma was used as the substrate in the measurement of plasma prothrombin activity. A control group consisting of five normal volunteers was included for comparison. Both the activity and concentration of prothrombin were significantly lower in the nephrotic group as compared with the control group. Significant quantities of immunoreactive prothrombin were detected in the urine of the majority of nephrotic patients. This study has provided unequivocal evidence of urinary excretion and acquired deficiency of prothrombin in the nephrotic syndrome.