Wanted: Dead or Alive Cells with Propidium Iodide Staining in Liver Tissue.

This study demonstrates the effectiveness of propidium iodide as a reliable marker for detecting dead or dying cells in frozen liver tissue sections. By comparing propidium iodide staining with the widely used Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, both methods showed consistent results in disease models such as alcohol-induced fibrosis and Western diet-induced fatty liver. Additionally, propidium iodide was successfully co-stained with other fluorescent markers, like phalloidin (for actin filaments) and antibodies targeting collagen, enabling detailed spatial analysis of dying cells within tissue.

Improved Syntheses and Some Selective Transformations of 2,2,4,4-Tetrachloro-8-oxabicyclo[3.2.1]oct-6-en-3-ones.

The methyl- and alkenyl-substituted furans 1b-h react with pentachloroacetone (2) and sodium 2,2,2-trifluoroethoxide solution to form the title compounds 3b-h in good yield. With the furans 1i-m bearing an oxygen or a sulfur heteroatom in the side chain, moderate yields are obtained. Dechlorination of the [4+3] cycloadducts 3 with the Zn-Cu couple leads to the corresponding oxabicyclic ketones 4.

Relationship between clinical stage, histopathology and antibody titers against the second Epstein-Barr virus nuclear antigen (EBNA-2) in non-Hodgkin's lymphoma patients.

Non-Hodgkin lymphoma (NHL) patients with centroblastic (Cb) or centroblastic-centrocytic (Cb/Cc)-diffuse lymphomas, immunocytoma (IC) and chronic lymphocytic leukemia (CLL) in clinical stages III-IV and with active disease (highly malignant group) were compared to NHL patients with CLL, IC, and centrocytic (Cc) or centroblastic-centrocytic (Cb-Cc)-diffuse/follicular lymphomas, in clinical stages I-II and with inactive disease (low malignant group) based on the presence of antibodies to Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA-1) and 2 (EBNA-2). In the highly malignant group, anti-EBNA-1 geometric mean titers (GMT) were 13.2 (range less than 2-80) and anti-EBNA-2 60.

Epstein-Barr virus (EBV) antigen-specific leukocyte migration inhibition in infectious mononucleosis. II. Kinetics of sensitization against five EBV-encoded nuclear proteins and the latent membrane protein.

The T cell-mediated immune response of infectious mononucleosis (IM) patients to five Epstein-Barr virus (EBV)-determined nuclear antigens, EBNAs, and to the membrane antigen associated with growth-transformed cells (latent membrane protein, LMP) was measured by the leukocyte migration inhibition (LMI) assay. Two different antigen sources were used: extracts from cells that only expressed EBNA-1, EBNA-2, or LMP after transfection with the corresponding EBV-DNA fragment, and synthetic peptides deduced from the corresponding genes. Patients in the acute phase of the disease failed to respond to EBNA-1, -5, -6, and LMP, but became responsive during convalescence.

Severe thrombocytopenia in Epstein-Barr virus-induced mononucleosis.

Severe thrombocytopenia is a rare complication of Epstein-Barr virus-induced infectious mononucleosis. We evaluated the clinical and laboratory data from seven patients seen between 1976 and 1985 whose lowest platelet counts varied from 3 to 25 x 10(9) per liter. Five of the seven patients were initially thought to have either acute leukemia or idiopathic thrombocytopenic purpura; eventually, however, primary Epstein-Barr virus infections were confirmed in all patients.

Heterophil-negative mononucleosis-like illnesses with atypical lymphocytosis in patients undergoing seroconversions to the human immunodeficiency virus.

The authors present data from four patients with acute heterophil-negative mononucleosis-like illnesses who were initially thought to have primary Epstein-Barr virus (EBV) infections but eventually were shown to be seroconverting to the human immunodeficiency virus (HIV). Widespread lymphadenopathy and blood smears indistinguishable from those typically encountered in the acute phase of infectious mononucleosis were present in all cases. There were also varying combinations of fever, sore throat, and malaise, as well as mild abnormalities of hepatic function and elevated cold agglutinins (anti-I).

Cellular and humoral immune responses to synthetic peptides deduced from the amino-acid sequences of Epstein-Barr virus-encoded proteins in EBV-transformed cells.

Ten synthetic peptides containing 18-22 residues deduced from the amino-acid sequences of the EBV-encoded latent-infection-associated membrane protein (LMP) and the 2 principal nuclear antigens, EBNA-1 and EBNA-2, were tested for their ability to induce lymphokine release from sensitized T-cells of EBV-seropositive donors, as measured by the leukocyte migration inhibition assay (LMI). Only one of the 10 free peptides induced EBV-specific LMI. After Sepharose-coupling, 4 additional peptides were regularly active.

Selected aspects of acute and chronic infectious mononucleosis and mononucleosis-like illnesses for the practicing allergist.

The diagnosis of EBV-IM or a heterophil-negative mononucleosis-like syndrome is best approached by combining morphologic and serologic data. The minimal hematologic criteria should always be searched for before accepting a case as IM or an IM-like illness. If minimal morphologic data are not rigidly adhered to, the number of heterophil-negative cases included under the umbrella of IM or an IM-like illness will swell and include a variety of other illnesses where early diagnosis may be important for treatment purposes.

Stable transfection of a human lymphoma line by sub-genomic fragments of Epstein-Barr virus DNA to measure humoral and cellular immunity to the corresponding proteins.

An Epstein-Barr virus (EBV)-negative human lymphoid B-cell line, DG75, was stably transfected with recombinant selection vectors that carry a subfragment of the BamHI WYH region (nucleotides 44664 to 50628), the BamHI K fragment, or a subfragment of the EcoRI D region (nucleotides 166614 to 170149) of B95-8 EBV DNA. These fragments contain the coding exons for the EBV-determined nuclear antigens EBNA2 and EBNA1, and the membrane antigen LMP, respectively. Antigen expression of the cells was detected by immunofluorescence.

Nuclear DNA-binding proteins determined by the Epstein-Barr virus-related simian lymphotropic herpesviruses H. gorilla, H. pan, H. pongo and H. papio.

Nuclear DNA-binding proteins were extracted from lymphoblastoid cell lines transformed with Epstein-Barr virus (EBV) or with the related lymphotropic herpesviruses of gorilla (Herpesvirus gorilla), chimpanzee (H. pan), baboon (H. papio) or orang-utan (H.

Marked hyperbilirubinemia in infectious mononucleosis. Analysis of laboratory data in seven patients.

While mild to moderate hepatic dysfunction is commonly encountered in infectious mononucleosis induced by Epstein-Barr virus (EBV), clinical jaundice with high bilirubin levels (greater than or equal to 6.0 mg/dL [greater than or equal to 103 mumol/L] is only occasionally encountered. In this study, seven patients with primary EBV infections had peak bilirubin levels of 10.

Characterization of EBV-carrying B-cell populations in healthy seropositive individuals with regard to density, release of transforming virus and spontaneous outgrowth.

Peripheral or tonsil lymphocyte populations of EBV-seropositive donors give rise to EBV-carrying LCLs upon in vitro explantation. Such lines can arise either by a 2-step mechanism, namely release of virus from some of the explanted cells followed by infection of previously uninfected B cells, or by direct outgrowth of virus-harboring B cells (Rickinson et al., 1974; Dalens et al.

Analysis of Epstein-Barr virus-specific and non-specific immune functions in a patient during the development of a non-Hodgkin's lymphoma.

A 57-year-old woman who presented with a reactive non-malignant lymphadenopathy was observed subsequently during the development of a nodular centroblastic non-Hodgkin's B-cell lymphoma. The Epstein-Barr virus (EBV)-specific antibody profile and EBV-specific and non-specific cell-mediated immune functions were determined at first presentation, and at various times during progression, in order to determine whether EBV was causally involved in the lymphoma and to assess in general the patient's cell-mediated immune function. At presentation, an immunodeficient status was suggested by an EBV-specific antibody profile indicative of an activated persistent infection; high antibody titers to viral capsid antigen (VCA) and early antigens (EA), but a low level of antibodies to EBV nuclear antigen (EBNA) confirmed by lack of leukocyte migration inhibition in response to EBNA (LMI-EBNA).

Antibody responses to Epstein-Barr virus-determined nuclear antigen (EBNA)-1 and EBNA-2 in acute and chronic Epstein-Barr virus infection.

Five distinct Epstein-Barr virus (EBV)-determined nuclear antigens (EBNA-1 to EBNA-5) were recently identified. Antibody responses to these antigens could conceivably differ, and thus prove of serodiagnostic value, in EBV-associated disease processes. As a first step, murine or human cell lines transfected with appropriate EBV DNA fragments and stably expressing either EBNA-1 or EBNA-2 were used to determine the frequency and time of emergence of antibodies to these two antigens in the course of acute and chronic infectious mononucleosis (IM) and to assess their titers in so-called chronic active EBV infections.

Epstein-Barr virus (EBV) antigen-specific leukocyte migration inhibition (LMI) in infectious mononucleosis (IM). I. Kinetics and response to a membrane protein on EBV-transformed cells.

Cell-mediated immune response of mononucleosis (IM) patients to Epstein-Barr virus (EBV)-determined antigens was measured by the leukocyte migration inhibition (LMI) assay. Patients in the acute phase of the disease failed to respond to partially purified nuclear antigen, EBNA, or to cell extracts that contained EBNA as the predominant EBV antigen. They showed a strong specific response to cell extracts enriched in early antigen (EA) and virus capsid antigen (VCA).

Absence of antibodies to the human immunodeficiency virus in sera from Africa prior to 1975.

Three different assays for detection of antibodies to the human immunodeficiency virus (HIV) were conducted on 677 sera obtained from 1964 to 1975 from male and female children and adults in Uganda and other countries in Africa. Several sera were collected from individuals with Kaposi sarcoma. No evidence of antibodies to the virus was noted up to 1975.

Attachment of antinuclear antibodies to nasopharyngeal carcinoma or other cells during preparation of biopsy imprints.

The presence of Epstein-Barr virus (EBV) genomes in nasopharyngeal and other carcinomas or Burkitt's and other B-cell lymphomas can be established by the demonstration of viral nucleic acid sequences in DNA extracts from biopsy specimens, the detection of EBV-associated nuclear antigen (EBNA) in biopsy imprints, and the inhibition of leukocyte migration by tumor extracts. Of these techniques, the detection of EBNA-positive tumor cells can be performed most readily in the laboratory. This report shows that a patient's antibodies to nuclear antigens can gain access to cell nuclei during the preparation of imprints.

Four virally determined nuclear antigens are expressed in Epstein-Barr virus-transformed cells.

The expression of Epstein-Barr virus (EBV)-determined antigens associated with growth-transformation of B cells was studied by immunoblotting with human sera from healthy donors. Four antigens were detected in EBV-carrying cell lines and in B lymphocytes early after infection with the transforming B95-8 substrain of virus. They were not found in uninfected cells, nor could they be demonstrated with sera lacking antibodies to EBV antigens.

Clinical and laboratory evaluation of cytomegalovirus-induced mononucleosis in previously healthy individuals. Report of 82 cases.

The present report describes the clinical and laboratory profile of 82 previously healthy individuals who developed cytomegalovirus (CMV)-induced mononucleosis. Many of these patients posed initial diagnostic problems and were hospitalized with diagnoses such as fever of undetermined origin, active viral hepatitis, acute leukemia, probable systemic lupus erythematosus, autoimmune hemolytic anemia, and severe pancytopenia. These patients underwent a variety of diagnostic biopsies, including liver biopsies (6) and bone marrow aspirations (9).

The Epstein-Barr virus determined nuclear antigen is composed of at least three different antigens.

The EBV-determined nuclear antigen, EBNA, is the only known viral product to be regularly detected in all EBV-transformed cells. The anticomplement immunofluorescence (ACIF) staining detects an EBV-specific nuclear reaction that has recently been shown to be due to at least 2 different proteins, EBNA-1 and EBNA-2, encoded by different parts of the viral genome. We now report the existence of a third antigen of the EBNA complex, designated as EBNA-3.

IgA antibodies to Epstein-Barr virus in infectious mononucleosis.

The IgA anti-EBV (Epstein-Barr virus) response during the course of IM (infectious mononucleosis) was investigated. The IgA anti-VCA (viral capsid antigen) response was found not to be restricted to the early acute phase of the EBV infection as is the IgM anti-VCA response. Some patients with normal total serum IgA levels did not respond with measurable EBV specific IgA.

Differences in EBV-specific antibody patterns at onset of infectious mononucleosis.

The EBV-specific antibody patterns of infectious mononucleosis (IM) patients were analyzed in relation to the onset of symptoms and clinical parameters during the acute phase of the disease. The antibody patterns varied considerably on admission. Three groups of patients were identified: one had not yet attained peak antibody titers, the second was at the peak and the third had passed the peak pattern.

Epstein-Barr virus infection and serological profile in Greenland Eskimo children.

The Epstein-Barr virus (EBV)-specific antibody profile of 101 Greenland Eskimo children was determined. The proportion of children with serological evidence of recent or past primary EBV infections rose from 22% at 6 months of age to 79% at 24 months of age. All but 2 of 49 children more than 4 years of age proved seropositive.

High prevalence of antibodies to acquired immune deficiency syndrome (AIDS)-associated retrovirus (ARV) in AIDS and related conditions but not in other disease states.

A rapid, sensitive indirect immunofluorescence assay has been developed for detection of antibodies to the acquired immune deficiency syndrome (AIDS)-associated retrovirus (ARV). The human T-cell line HUT-78 was chronically infected with ARV-2 and used to detect antibodies to virus-specific cytoplasmic antigens. Because the helper T-cell marker Leu-3 is substantially reduced in this cell line after ARV infection, it appears to be an important receptor for virus infection.