howard reisner - Academia.edu (original) (raw)

Papers by howard reisner

European Journal of Oral Sciences, 1999

Due to their peripheral location in the dental pulp and their cellular extension into dentin, odo... more Due to their peripheral location in the dental pulp and their cellular extension into dentin, odontoblasts are the ®rst pulpal cells to encounter dental pathogens. The association of odontoblasts with immunoglobulins and dendritic cells during microbial invasion of dentin implies that these cells may possess a role in the innate and adaptive pulpal immune responses, however this has not been examined. A pivotal step in the innate immune response is the detection of foreign antigen and the recruitment of immune effector cells to the area. IL-8 is a potent chemotactic cytokine that plays an important role in the in¯ammatory response. The purpose of this study was to determine if odontoblasts are capable of expressing the pro-in¯ammatory chemokine IL-8. Human odontoblasts from intact, noncarious third molars were maintained in culture and exposed to Escherichia coli lipopolysaccharide (LPS) (serotype 055:B5) on day 4 for 8±10 h in a humidi®ed 5% CO 2 incubator. Control and experimental samples were assayed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot for the production of IL-8 mRNA and protein. Analysis of the PCR products revealed that cells of the odontoblast layer maintained in this culture model constitutively expressed low levels of IL-8, which were increased in response to E. coli LPS exposure. Western blotting con®rmed that the mRNA was translated into protein. These results imply that odontoblasts are capable of producing of pro-in¯ammatory mediators, thereby actively participating in the recruitment of neutrophils in response to bacterial by-products.

Blood, 1988

Two men with factor IX (FIX)antigen-positive (CRM+) hemophilia B were selected for study because ... more Two men with factor IX (FIX)antigen-positive (CRM+) hemophilia B were selected for study because of their abnormal expression of an immunologically defined epitope previously localized to the EGF-like domains of the molecule. Exons IV and V (coding for the first and second EGF-like domains) of FIX were amplified 10(7) times from the…

Proceedings of the National Academy of Sciences, 1989

We have used the polymerase chain reaction to amplify the entire coding region of canine factor I... more We have used the polymerase chain reaction to amplify the entire coding region of canine factor IX from a hemophilia B animal. When the sequence was compared to that which codes for normal canine factor IX, a single missense mutation was identified. This mutation (G----A at nucleotide 1477) results in the substitution of glutamic acid for glycine-379 in the catalytic domain of the molecule. The mutation creates a new restriction site that allowed confirmation of the abnormal sequence in both hemophilic and carrier animals. Amino acid 379 in canine factor IX corresponds to position 381 in human factor IX, a location at which no human mutations have been described. Moreover, it occurs at one of the few amino acids that have been rigorously conserved among the trypsin-like serine proteases throughout evolution. The mutation responsible for canine hemophilia B results in a complete lack of circulating factor IX in the affected animals. As it is unusual for a missense mutation to result ...

British Journal of Haematology, 1978

A visual assay of factor VIII-related Willebrand factor (VIIIR:WF) is described which utilizes fo... more A visual assay of factor VIII-related Willebrand factor (VIIIR:WF) is described which utilizes formaldehyde-fixed platelets, end points being read in microflocculation tiles. Four dilutions of a sample can be assessed simultaneously, and the correlation with aggregometric assays is high (r = 0.91). Measurement error is 8.0% for a single assay in triplicate and less than 5% if an assay is repeated three times. The method has been used for 2 years by the coagulation genetics group at Chapel Hill for diagnosing subjects with von Willebrand's disease and assigning genotypes to members of families transmitting this disorder. Its utility in classifying known carriers of haemophilia A has also been examined, both in conjunction with assays of VIII:C and in a three-way test with assays of VIII:C and VIIIR:Ag. As predicted by the Lyon hypothesis, the rate of false negative diagnosis was higher than false positive diagnosis, but the overall rate of misclassification on single plasma samples was 7/51 = 13.7%. The error rate was the same whether discrimination was based upon assays of VIII:C vs. VIIIR:Ag, VIII:C vs. VIIIR:WF, or VIII:C vs. VIIIR:Ag vs VIIIR:WF, the same individuals being misclassified by each method. The observed rate of misclassification was well within the rates reported by others and very similar to our previous experience. We have concluded that this method of assaying VIIIR:WF is highly useful for diagnosing vWd, detecting inhibitors to VIIIR:WF, and examining large numbers of column fractions. It is a useful supplement, although it cannot yet substitute for, assays of VIIIR:Ag in detecting carriers of haemophilia A.

We used recombinant DNA techniques to map a panel of six monoclonal antibodies (MoAbs) to regions... more We used recombinant DNA techniques to map a panel of six monoclonal antibodies (MoAbs) to regions of the human factor IX molecule. A-2 maps to 1 7 amino acids at the amino terminus of the heavy chain of IXa; 2D5. an inhibitor of clotting. is defined to 36 amino acids of the first EGF-like domain of human factor IX. A-4. A-5. ClOD, and FXCOO8 all F UNCTIONAL FACTOR IX is essential for normal hemostasis. A defective factor IX molecule results in hemophilia B, an X-linked hereditary bleeding disorder. Human factor IX has a molecular weight (mol wt) of-57,000 daltons and contains ‘-.17 % carbohydrate.”2 The first 12 amino terminal glutamic acid residues of the zymogen are modified to ‘y-carboxyglutamic acids (Gla) in a posttranslational vitamin K-dependent reaction.2’5 These Gla residues are required for function and are responsible for most of the calcium-dependent binding of factor IX to

Transplantation, 2014

Background. A qualitative highly predictive urinary test for polyomavirus nephropathy (PVN) is th... more Background. A qualitative highly predictive urinary test for polyomavirus nephropathy (PVN) is the PV-Haufen test. This article evaluates whether a quantitative PV-Haufen analysis, that is, the number of PV-Haufen shed per milliliter urine, predicts PVN disease grades and the severity of intrarenal PV replication. Methods. Polyomavirus-Haufen were counted in 40 urine samples from patients with biopsy-proven definitive PVN. The number of PV-Haufen was correlated with both histologic PVN disease grades 1 to 3 and the number of SV40-T-expressing cells as indicators of intrarenal PV replication in corresponding renal allograft biopsies (manual counts and automated morphometry). Findings from quantitative PV-Haufen analyses were compared to conventional laboratory test results, that is, BK viremia (quantitative polymerase chain reaction [PCR]) and BK viruria (quantitative PCR and decoy cell counts). Results. Polyomavirus-Haufen counts showed excellent correlation (α0.77-0.86) with the severity of intrarenal PV replication and disease grades. In particular, low PV-Haufen numbers strongly correlated with early PVN grade 1 and minimal intrarenal expression of SV40-T antigen (P < 0.001). In comparison, BK viremia and viruria levels by PCR showed only modest correlations with histologic SV40-T expression (α0.40-0.49) and no significant correlation with disease grades or minimal intrarenal PV replication. No correlations were seen with urinary decoy cell counts. In contrast to conventional quantitative PCR assays or decoy cell counts, quantitative urinary PV-Haufen testing accurately reflects the severity of PV replication, tissue injury, and PVN disease grades. Conclusions. Quantitative PV-Haufen testing is a novel noninvasive approach to patient management for the diagnosis and prediction of PVN disease grades and monitoring of disease course during therapy.

XIth International Congress on Thrombosis and Haemostasis

In order to map the regions of human factor IX recognized by monoclonal antibodies we have insert... more In order to map the regions of human factor IX recognized by monoclonal antibodies we have inserted random fragments of the coding region of the cDNA for human factor IX into the lambda phage expression vector, lambda gtll. The resultant recombinant phage were screened with monoclonal antibodies, the immunoreactive phage were isolated, and the DNA of the inserted fragment was sequenced to determine which amino acids were immunoreactive. This data, coupled with data obtained from the use of specific fragments of human factor IX expressed in E. coli from a T7 phage promoter, has allowed us to map the location of several epitopes on the surface of the factor IX molecule. In those cases where the antibodies are specific for human factor IX, additional narrowing of the epitope is possible by comparing the amino acid sequence of human factor IX to the bovine molecule. Six monoclonal antibodies from 3 different laboratories have been mapped. The immunodominant epitopes appear to be the ami...

VIIth International Congress on Thrombosis and Haemostasis

Somatic cell hybrids have been produced using primary cultures of human vascular endothelial cell... more Somatic cell hybrids have been produced using primary cultures of human vascular endothelial cells and rodent cell strains. Production of factor VIII related antigen (VIIIR :Ag) by the cultured endothelial cells can be clearly demonstrated by a direct fluorescent antibody assay, but has not yet been detected in 10 of 10 hybrid clones. The hybrid cells do express many other human traits. Isozyme analyses for human chromosome markers demonstrate that various human chromosomes have been segregated from each of the hybrids. There are several tenable explanations for the fact that human VIIIR:Ag has not been detected in these hybrid cells. 1) Factor VIII may require a human chromosome(s) not present in any of the hyhrids ana1yzed to date. 2) Factor VIII may be expressed at lower levels in hybrid cells such that it would be detectable only by a more sensitive assay. 3) The rodent genome may block expression of this differentiated function in the hybrid cells. Since in a whole organism, on...

Http Www Libreriasaulamedica Com, 2013

Journal of Endodontics, 1999

Haemostasis

The levels of factors VIII, II, and VII rise during pregnancy in normal women. In addition, incre... more The levels of factors VIII, II, and VII rise during pregnancy in normal women. In addition, increases in factor VIII levels have been observed in pregnant carriers of hemophilia A and in women affected with von Willebrand's disease. The influence of pregnancy on factor IX levels is less clear. Consequently, we determined serial factor IX coagulant activities (IX:C) and factor IX antigen levels (IX:Ag) during the pregnancy of an affected carrier of hemophilia B who had baseline values of 13% both IX:C and IX:Ag levels. Neither level rose during pregnancy and the patient was treated with plasmapheresis and plasma infusions during the delivery and the postpartum period. Excessive bleeding did not occur.

The American Journal of Human Genetics

We have described the study of a small kindred with X-linked hemophilia A. It was ascertained thr... more We have described the study of a small kindred with X-linked hemophilia A. It was ascertained through a clinically affected female, the daughter of a man with moderately severe hemophilia. The pedigree and the proband's phenotype suggest that she may be a heterozygote in whom most of the normal alleles at the VIII-1 locus are not active. She has two sisters, also obligatory carriers. The three sisters exhibit the three phenotypes possible for heterozygous females: clinically affected, clinically normal but phenotypically abnormal as determined by laboratory tests, and clinically and phenotypically normal.

Laboratory investigation; a journal of technical methods and pathology, 1996

Anti-neutrophil cytoplasmic autoantibodies (ANCA) have been hypothesized to participate in the pa... more Anti-neutrophil cytoplasmic autoantibodies (ANCA) have been hypothesized to participate in the pathogenesis of necrotizing vasculitis based on their association with small vessel vasculitides and the in vitro ability of such antibodies to activate cytokine-primed neutrophils. Much remains to be elucidated about the factors responsible for the generation and perpetuation of these autoantibodies and the shaping of the ANCA immune response. This study evaluated the clonal diversity of the ANCA immune response in patients with myeloperoxidase-ANCA associated disease. Isoelectric focusing was used to investigate the clonality of myeloperoxidase-ANCA from 34 patients with pauci-immune necrotizing glomerulonephritis. Sixty-nine percent of the patients had two or less clonotypes to myeloperoxidase, whereas 31% had more than two clonotypes. Clonality was stable over the course of the disease and shared among some unrelated patients. Shared idiotypy was specifically investigated using a murin...

Thrombosis and haemostasis, 1996

Hemophilia A is a clotting disorder that is due to reduced or absent coagulation factor VIII (FVI... more Hemophilia A is a clotting disorder that is due to reduced or absent coagulation factor VIII (FVIII) activity. In approximately 25% of people with severe hemophilia A, standard treatment with intravenous plasma-derived or recombinant FVIII (rFVIII) induces anti-FVIII antibodies that inhibit FVIII activity (inhibitors). We describe the development of a rat model to study the formation of inhibitors. Immunization of rats with human rFVIII in adjuvant induced an anti-human rFVIII antibody response characteristic of an anti-FVIII inhibitor response in hemophilia A patients. The rats exhibited a rapid, polyclonal secondary antibody response to human rFVIII. These antibodies were reactive against epitopes located in the heavy and light chains. All the rFVIII-immunized rats developed antibodies against the FVIII C2 domain, a region of major reactivity in hemophilia A patients with inhibitors. Furthermore, competition ELISAs demonstrated that rat and human anti-FVIII antibodies recognized i...