pkl1(+)and klp2(+): Two kinesins of the Kar3 subfamily in fission yeast perform different functions in both mitosis and meiosis.

We have identified Klp2p, a new kinesin-like protein (KLP) of the KAR3 subfamily in fission yeast. The motor domain of this protein is 61% identical and 71% similar to Pkl1p, another fission yeast KAR3 protein, yet the two enzymes are different in behavior and function. Pkl1p is nuclear throughout the cell cycle, whereas Klp2p is cytoplasmic during interphase. During mitosis Klp2p enters the nucleus where it forms about six chromatin-associated dots. In metaphase-arrested cells these migrate back and forth across the nucleus. During early anaphase they segregate with the chromosomes into two sets of about three, fade, and are replaced by other dots that form on the spindle interzone. Neither klp2(+) nor pkl1(+) is essential, and the double deletion is also wild type for both vegetative and sexual reproduction. Each deletion rescues different alleles of cut7(ts), a KLP that contributes to spindle formation and elongation. When either or both deletions are combined with a dynein deletion, vegetative growth is normal, but sexual reproduction fails: klp2 Delta,dhc1-d1 in karyogamy, pkl1 Delta,dhc1-d1 in multiple phases of meiosis, and the triple deletion in both. Deletion of Klp2p elongates a metaphase-arrested spindle, but pkl1 Delta shortens it. The anaphase spindle of klp2 Delta becomes longer than the cell, leading it to curl around the cell's ends. Apparently, Klp2p promotes spindle disassembly and contributes to the behavior of mitotic chromosomes.

Absence of nasal mucosal atrophy with fluticasone aqueous nasal spray.

OBJECTIVE: To evaluate whether 1 year of continuous treatment with intranasal fluticasone propionate would lead to atrophy in the nasal mucosa compared with an active control, oral terfenadine. DESIGN: Prospective, randomized, multicenter, open-label, parallel-group study. SETTING: Two tertiary care academic institutions. PATIENTS: Seventy-five subjects older than 18 years with perennial allergic rhinitis. INTERVENTIONS: Patients received either fluticasone propionate aqueous nasal spray, 200 microg once daily, or terfenadine, 60 mg twice daily, for 1 year. Nasal biopsy specimens were obtained before and after 1 year of treatment and were evaluated for evidence of atrophy. MAIN OUTCOME MEASURES: Epithelial and collagen layer thickness of the nasal mucosa as assessed by light microscopy and the presence and degree of edema, and regularity of collagen fibrils as assessed by electron microscopy. Analyses were performed without knowledge of subject identity or treatment assignment. RESULTS: Neither fluticasone nor terfenadine treatment led to atrophy in the nasal mucosa by clinical or histologic observation. No significant changes from baseline were observed for any assessment of atrophy. In contrast to what would have been expected if atrophy were to occur, mean epithelial layer thickness in the fluticasone group significantly increased compared with terfenadine treatment (P = .03). CONCLUSIONS: Treatment with intranasal fluticasone for 1 year increases the thickness of the nasal epithelium as compared with a year's treatment with terfenadine and does not lead to atrophy in the nasal mucosa. The increased thickness in the fluticasone treatment may represent repair from epithelial damage caused by chronic allergic inflammation.

Necrotizing granulomatous cervical lymphadenitis associated with B lymphocyte abnormalities and a positive PPD reaction.

Sarcoidosis is a diagnosis often made by exclusion. We present a case of what we believe is sarcoidosis presenting as cervical lymphadenopathy. Complicating issues in this patient included B lymphocyte abnormalities, and a positive PPD reaction. The patient continues to be steroid dependent. This case illustrates the difficulties in diagnosis and management of sarcoidosis.

Comparison of four immunoserologic assays for detection of antibodies to Borrelia burgdorferi in patients with culture-positive erythema migrans.

In view of the significant sequelae associated with Lyme borreliosis, there is a need for timely and accurate diagnosis of erythema migrans (EM). Although Borrelia burgdorferi can be cultured from biopsies of EM lesions, immunodiagnostic testing is more widely available. Four immunoserologic methods were studied by using the sera of 51 patients with EM lesions that were culture positive for B. burgdorferi. Nineteen patients had single primary lesions, and thirty-two had multiple secondary lesions. At the time of biopsy, 40 patients, 8 with primary lesions and all patients with secondary lesions, were seropositive by an immunoglobulin M (IgM) indirect fluorescent-antibody (IgM IFA) test (Bion Enterprises). Twenty-three patients were seropositive by a whole-cell fluorescence enzyme immunoassay (EIA) (BioWhittaker, Inc.), twenty-two were positive by immunoblotting (ViroStat, Inc.), and one was positive by a P39 recombinant EIA (P39 EIA) (General Biometrics, Inc.). Sera from various patient control groups were tested: rheumatoid arthritis (n = 19), infectious mononucleosis (n = 20), systemic lupus (n = 22), syphilis (n = 13), streptococcal sequelae (n = 20), and healthy subjects (n = 16). None of these sera reacted with the IgM IFA test or P39 EIA. Fifteen reacted with the fluorescence EIA. We conclude that the IgM IFA test is an effective and reliable assay for the diagnosis of EM, particularly for patients with secondary lesions. Immunoblot, fluorescence EIA, and P39 EIA lack the sensitivity to reliably diagnose EM.

Evaluation of the impact of intraobserver variability on endometrial dating and the diagnosis of luteal phase defects.

OBJECTIVE: To determine the magnitude of intraobserver variation in dating endometrial biopsies and its impact on clinical management. DESIGN: Blinded histopathologic interpretation of endometrial biopsy specimens 1 year apart by five pathologists. SETTING: Large military tertiary care center. PATIENTS: Endometrial biopsy specimens from 51 patients undergoing evaluation for potential luteal phase defects. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Calculation of the magnitude of the individual and overall intraobserver variation in endometrial dating for the five pathologists and estimation of its potential impact on clinical management. RESULTS: The intraobserver variation was 0.69 +/- 0.05 days (means +/- SE). There was no significant difference in the magnitude of the variation for 1-day or 2-day dating ranges. The theoretical probability of altering clinical management by having the same pathologist redate a given specimen ranged from 15% to 28%. CONCLUSION: Histologic dating of endometrial biopsies is subject to a small but highly clinically significant intraobserver variability that may have a major impact on clinical management.

Primary and secondary erythema migrans in central Wisconsin.

BACKGROUND AND DESIGN: We report a series of 28 consecutive patients with erythema migrans (EM) who underwent skin biopsies for culture of Borrelia burgdorferi. Culture results, Lyme serologic findings, and clinical features in patients with only primary EM are compared with those in patients with secondary EM. RESULTS: Culture preparations of skin specimens obtained from six of 12 patients with only primary EM, and from 14 of 16 patients with secondary EM were positive for B burgdorferi. Seven patients with only primary EM were initially seronegative, and only one patient had an annular lesion. A central crusted punctum was present in five of six primary EM lesions that were not culture positive, but in none of nine culture-positive primary EM lesions. Patients with secondary EM were all seropositive and had less cutaneous and more constitutional symptoms than patients with only primary EM. Three patients with secondary EM had abnormal liver enzyme profiles, one patient had complete heart block, and one patient had myocarditis simulating infarction. Less than one third of all patients recalled a tick bite. An isomorphic nonresponse was seen in skin previously involved with secondary EM in one patient who had a drug exanthem from amoxicillin. CONCLUSIONS: Borrelia burgdorferi can be reliably cultured from skin biopsy specimens of secondary EM. Culture preparation aids definitive diagnosis of Lyme disease in patients with only primary EM who often lack constitutional symptoms, have nondiagnostic lesions, or are seronegative.

Isolation of Borrelia burgdorferi from skin biopsy specimens of patients with erythema migrans.

Procedures for the cultural isolation and identification of Borrelia burgdorferi from skin biopsy specimens are described. B. burgdorferi was isolated from 24 of 34 skin biopsy specimens from patients with erythema migrans. Eight of the culture-positive patients had single, primary lesions and 16 had multiple, secondary lesions. The 17 male and 7 female patients were 2 to 70 years old. Biopsy samples were obtained from erythematous or normal-appearing skin within 1 cm of the peripheral aspect of the lesion. Twenty-three of the isolates were detected within 8 days of incubation in Barbour-Stoenner-Kelly medium with no antimicrobial agents. The identities of the isolates were determined by reactivity with monoclonal antibodies H9724 and H5332. Cultivation of B. burgdorferi from skin lesions suggestive of erythema migrans is a practical and clinically relevant procedure. Clinical isolates and corresponding patient sera and urine will contribute to efforts to improve existing immunoserologic testing methods and develop new assays to diagnose Lyme borreliosis.

Intranuclear inclusions correlate with the ultrastructural detection of herpes-type virions in oral hairy leukoplakia.

Hairy leukoplakia (HL) is an Epstein-Barr virus-associated lesion of the oral mucosa that occurs in persons infected with human immunodeficiency virus. Ultrastructural studies have demonstrated herpes-type virions in 63-100% of cases of HL. In this investigation, incisional biopsy specimens from 52 consecutive patients with oral lesions clinically suspicious for HL were analyzed by light and electron microscopy to determine which light microscopic features correlated best with the presence of herpes-type virions ultrastructurally. The predominant histopathologic features consisted of irregular hyperparakeratosis, acanthosis, koilocytoid cells, and an absence of significant inflammation. Candidal organisms and bacterial colonies were variable findings. Intranuclear inclusions, identical to Cowdry type A inclusions seen in other herpesvirus infections, were observed in 46 cases and demonstrated the best correlation with the presence of virions ultrastructurally (p less than 0.0001). Koilocytoid cells also had a statistically significant correlation with the presence of virions (p less than 0.001). We conclude that Cowdry type A inclusions are present in many cases of HL and that their presence, when combined with the other clinical and histologic findings in these lesions, is the most specific light microscopic feature of HL.

Ultrastructural detection of herpes-type virions by negative staining in oral hairy leukoplakia.

Hairy leukoplakia (HL) is a newly recognized virus-associated lesion of oral mucosa that occurs in persons infected with human immunodeficiency virus. Studies have demonstrated Epstein-Barr virus within epithelial cells of HL. The authors examined 12 cases of HL by transmission electron microscopy to compare the use of a negative staining technique versus routine plastic embedment for the detection of viruses. Herpes-type virions were identified by both methods in 11 cases. One case had negative results for viruses by both methods. Negative staining is a simple and rapid technique that compares favorably with plastic embedment in the detection of herpes-type virions in HL.

Abnormal lung and liver uptake of gallium-67 and technetium-99m MDP in hypercalcemia of lymphoma with metastatic pulmonary calcification.

Abnormal pulmonary uptake of Ga-67 citrate and Tc-99m MDP and reversible liver uptake of Tc-99m MDP was seen in a patient with hypercalcemia of lymphoma and biopsy-proven metastatic pulmonary calcification. Abnormal lung uptake of Tc-99m MDP may confirm the diagnosis of pulmonary calcification, lessening the need for invasive procedures to evaluate pathologic lung uptake of Ga-67 citrate.

Massive intravascular hemolysis without serum haptoglobin depletion.

Hyperplasia of marrow histiocytes with extensive hemophagocytosis was found in a patient with cytomegalovirus infection. He experienced massive intravascular hemolysis, but, unexpectedly, no depression of the serum haptoglobin level was found by either single radial immunodiffusion or rate nephelometry. The unexpectedly high haptoglobin value may have been the result of "blockade" of the monocyte-macrophage system, with resultant failure to clear haptoglobin-hemoglobin complex rapidly from the circulation. Use of the techniques described for the indirect estimation of unbound serum haptoglobin alone should avoid confusing results in similar clinical circumstances.