Francis W. Chandler

Immunohistochemical evaluation of bcl-2 oncoprotein in oral dysplasia and carcinoma

The proto-oncogene bcl-2 is associated with follicular lymphoma involving translocation t(14;18)(q32;q21) and is also overexpressed in various neoplasms. We report deregulation of bcl-2 expression during progression from oral epithelial dysplasia to squamous cell carcinoma. Immunohistochemical analysis with monoclonal antibodies to bcl-2 oncoprotein in formalin-fixed paraffin-embedded tissue sections revealed that severe epithelial dysplasias had a higher percentage of immunoreactivity than did mild and moderate dysplasias and squamous cell carcinomas. Expression of this oncoprotein was directly proportional to the degree of epithelial dysplasia, and nondysplastic basal cells contiguous to neoplastic lesions also expressed bcl-2. These findings, along with down-regulation of bcl-2 in differentiating carcinomas, suggest a role for this oncoprotein in relatively early stages of oral tumor progression. Differentiating neoplastic cells with marginal or no bcl-2 reactivity showed heterogeneous cell labeling of varying intensity for differentiation-associated cytokeratin (CK13), indicating their inverse topographic relationship.

Chlamydia trachomatis in chronic abacterial prostatitis: Demonstration by colorimetric in situ hybridization

Twenty-three transurethral resected prostate specimens with histologic evidence of chronic abacterial prostatitis were evaluated by colorimetric in situ hybridization for evidence of Chlamydia trachomatis. Intracellular chlamydia bodies were detected in seven of 23 cases (30.4%). In five of seven positive cases, chlamydia inclusion bodies were present within both epithelial cells and intraluminal histiocytes. In two of seven positive cases, extracellular chlamydia bodies were identified in conjunction with intraepithelial inclusions. In only one case were extraductal, extraacinar, intrahistiocytic inclusion bodies observed. Our study confirms previous reports implicating C trachomatis as an etiologic agent in chronic abacterial prostatitis, and underscores the applicability of DNA probes for the detection and identification of C trachomatis in routinely fixed, paraffin-embedded tissues.

Progressive facial hemiatrophy (Parry-Romberg syndrome) and borreliosis

We report a 5 year-old girl with progressive facial hemiatrophy and with borreliosis as suggested cause .

Detection of Borrelia burgdorferi in Human Blood and Urine Using the Polymerase Chain Reaction

We investigated the use of the polymerase chain reaction (PCR) to detect Borrelia burgdorferi strain B-31 in human blood and urine experimentally inoculated with 5 and 1 borreliae/cm3, respectively, and to biotinylate a DNA probe specific for B. burgdorferi in the dot blot and Southern blot assays. When the blood and urine samples were subjected to PCR, a 370-bp amplified product was consistently visible on agarose gel electrophoresis after 30 and 45 cycles, respectively. The total human genomic DNA extracted from a 1-cm3 sample of inoculated blood was approximately 6.25 micrograms, and the total amount of B. burgdorferi DNA was estimated to be 0.01 pg/6.25 micrograms of the human DNA. For PCR, 2.5 micrograms of human DNA which contained the equivalent of 0.004 pg of borrelia DNA (approximately two borreliae) were used for enzymatic amplification. When 1/20 or 1/10 of the PCR-amplified products were used either for dot blot or Southern blot hybridization, the accessible copies of amplified B. burgdorferi DNA were sufficient for detectable hybridization to occur. PCR amplification of B. burgdorferi DNA in clinical specimens followed by dot blot hybridization may be a valuable adjunct or alternative to current but inadequate laboratory methods for the diagnosis of Lyme disease.

Sensitization to recombinant Aspergillus fumigatus allergens in allergic fungal sinusitis

BACKGROUND Allergic bronchopulmonary mycosis is primarily caused by Aspergillus fumigatus. Despite similarities, allergic fungal sinusitis (AFS) is thought to be caused by various fungi. OBJECTIVE Identify fungal elements in AFS allergic mucin and determine the prevalence of specific immunoglobulin (Ig)E to recombinant A. fumigatus allergens (rAsp) in AFS patients. METHODS Allergic mucin from 17 definitive and 10 probable AFS patients were histologically examined for fungal elements. Sera from 18 definitive AFS patients, 10 probable AFS patients, 6 chronic sinusitis patients, and 5 A. fumigatus-allergic patients were tested for specific IgE to A. fumigatus and five rAsps. RESULTS Ten of the 17 definitive cases had hyphae morphologically resembling Aspergillus or Fusarium spp. One probable AFS patient had similar findings. Of definitive patients, 94% (17 of 18) showed A. fumigatus-specific IgE (> or = 0.35 kUa/L), and 67% were positive to one or more rAsp. Four of 10 probable patients demonstrated A. fumigatus-specific IgE, and 2 had IgE to one or more rAsp. The definitive group had greater mean A. fumigatus IgE (P < 0.05) versus the probable and chronic sinusitis groups. The definitive groups rate of IgE to the rAsps was statistically greater. All definitive patients with Aspergillus or Fusarium spp. in situ had A. fumigatus-specific IgE, and 7 of 10 had IgE to at least one rAsp. CONCLUSIONS Most definitive AFS patients have A. fumigatus-specific IgE and many have specific IgE to rAsps. Many also demonstrate Aspergillus spp. or Fusarium spp. in situ. Findings suggests that A. fumigatus is an important causative agent in AFS in the southeast United States.

Molecular Detection of JC Virus in Embalmed, Formalin-Fixed, Paraffin-Embedded Brain Tissue

Embalmed tissues are adequate for the detection of JC virus in lesions of progressive multifocal leukoencephalopathy (PML) by immunohistologic and molecular methods. JC virus was readily detected in embalmed brain tissue using immunohistochemistry (IHC), in situ hybridization (ISH), and the polymerase chain reaction (PCR). Two brains were removed from bodies that had been embalmed at least 24 h prior to autopsy. They were subsequently post fixed in 10% buffered formalin for 10-14 days before dissection and molecular studies were performed. Though these techniques are not novel, their use in embalmed tissues is. Routine embalming should not eliminate these diagnostic procedures from consideration.

Epidemiology of AIDS and its Opportunistic Infections

The acquired immunodeficiency syndrome (AIDS), now known to be caused by a novel retrovirus known as the human immunodeficiency virus (HIV) (Barre-Sinoussi et al., 1983; Gallo et al., 1984), is characterized by a profound derangement in cell-mediated immunity leading to multiple opportunistic infections and unusual neoplasms. AIDS was first recognized in the spring of 1981 when Pneumocystis carinii pneumonia (PCP) and Kaposi’s sarcoma (KS) appeared in previously healthy homosexual men in California and New York (CDC, 1981a,b). It soon became apparent that other opportunistic diseases were present in addition to PCP and KS, and that groups other than homosexual men were also at risk including intravenous (IV) drug users, recipients of blood and blood products, and heterosexual contacts of persons at increased risk (CDC, 1981c; CDC, 1982a). By autumn of 1981, the CDC had created a task force to conduct epidemiologic, clinical and laboratory investigations on AIDS. For purposes of national surveillance, a case definition of AIDS was developed in 1982 that required the diagnosis of one or more of certain diseases at least moderately predictive of a defect in cell-mediated immunity in persons without underlying conditions known to cause immunodeficiency, but did not generally require laboratory confirmation of either immunodeficiency or infection with HIV (CDC, 1982b).

Anemia in a patient with panhypogammaglobulinemia.

HISTORY OF PRESENT ILLNESS A 12-year-old African-American boy with panhypogammaglobulinemia, was referred to the Medical College of Georgia for further evaluation of a decreasing hematocrit and abdominal pain. The patient initially presented to a local hospital approximately two days earlier with generalized malaise, vomiting, dehydration, and severe abdominal pain. The abdominal symptoms improved with intravenous fluids and gut rest. He was placed on antibiotics due to radiographically documented sinusitis and pneumonia. His initial hematocrit was 24% and it later fell to 17%. He was then transferred to the Medical College of Georgia for further evaluation.