Antibodies against Crandell Rees feline kidney (CRFK) cell line antigens, alpha-enolase, and annexin A2 in vaccinated and CRFK hyperinoculated cats.

BACKGROUND: Cats inoculated with feline herpesvirus 1, calicivirus, and panleukopenia (FVRCP) vaccines grown on the Crandell Rees feline kidney (CRFK) cell line have been shown to develop anti-CRFK antibodies. The identities of common CRFK antigens are unknown. HYPOTHESIS: Cats inoculated with CRFK lysates and FVRCP vaccines will develop autoantibodies measurable by Western blot immunoassay. Antigens associated with these antibodies can be isolated for further study. ANIMALS: One CRFK hyperinoculated rabbit, 44 age-matched unvaccinated kittens purchased from a commercial vendor. METHODS: Commonly recognized CRFK antigens were identified by comparison of Western blot immunoassays using sera from a hyperinoculated rabbit and kittens inoculated with CRFK lysate or 1 of 4 commercially available FVRCP vaccines. Antigens were purified from CRFK lysates and sequenced. Antigen recognition was confirmed by Western blot immunoassay and indirect ELISA for 2 proteins using sera from CRFK and FVRCP inoculated kittens. RESULTS: CRFK antigens 47, 40, and 38 kD in size were identified. Protein isolation and sequencing identified 3 CRFK proteins as alpha-enolase, annexin A2, and macrophage capping protein (MCP). Sera from FVRCP and CRFK inoculated cats were confirmed to recognize annexin A2 and alpha-enolase by Western blot immunoassay and indirect ELISA. CONCLUSIONS AND CLINICAL RELEVANCE: This study validated the use of Western blot immunoassay for detection of antibodies against CRFK proteins and identified 3 CRFK antigens. In humans, alpha-enolase antibodies are nephritogenic; alpha-enolase and annexin A2 antibodies have been associated with autoimmune diseases. Further research will be necessary to determine the clinical relevance of these findings.

The hetero-oligomeric antigen receptor complex and its coupling to cytoplasmic effectors.

T-cell and B-cell antigen receptors are representative of a family of multisubunit receptors that utilize Src-family kinases as proximal cytoplasmic effectors in signal transduction. Recent studies have shown that distinct receptor subunits mediate ligand and effector interactions and demonstrate that physical interaction with effectors, and their activation, is a function of a 26 amino acid motif found in multiple receptor subunits. Further, receptor ligation induces tyrosine phosphorylation of this motif, and this initiates SH2-mediated association and activation of Src-family kinases and, apparently, ZAP70 kinases. Finally, this association triggers SH3-mediated binding of Lyn and Fyn to PI3-K, resulting in PI3-K activation. An integrated model of signal transduction is presented.

Thermodynamic analysis of the stabilisation of pig heart mitochondrial malate dehydrogenase and maize leaf phosphoenolpyruvate carboxylase by different salts, amino acids and polyols.

As part of our investigations into the inactivation of pig heart mitochondrial malate dehydrogenase (phm-MDH) and maize leaf phosphoenolpyruvate carboxylase (ml-PEPC) in the presence of various cosolvents, the denaturation kinetics as a function of temperature have been determined based on Arrhenius plots derived from transition state theory analysis over the temperature range from 3.5 degrees C to 65 degrees C. The experimental data for phm-MDH were obtained in the presence of 1 M concentrations of various salts of monovalent and polyvalent anions, 1 M amino acids or 1 M sucrose and 6.1 M glycerol. Similarly, Arrhenius plot data were obtained for ml-PEPC in the presence of 2.5 M NaOAc and 0.8 M sodium glutamate. Distinct regimes of inactivation corresponding to high and low values of inactivation enthalpy were identified for the phm-MDH in the presence of all cosolvents and for the ml-PEPC in the presence of 2.5 M NaOAc, but not in the presence of 0.8 M sodium glutamate. A significant temperature-dependent effect dominated the inactivation of phm-MDH and ml-PEPC at elevated temperatures (e.g., > or = 45 degrees C), whilst the inactivation of these enzymes over a lower temperature range (< or = 25 degrees C) was dominated by temperature-independent phenomenon. The corresponding thermodynamic activation parameters (deltaG++, deltaH++ and deltaS++) associated with the transition state complexes involved in the inactivation of phm-MDH and ml-PEPC in the presence of the various cosolvents have been determined. The results indicate that the transition states associated with the inactivation of these two enzymes at elevated temperatures are characterised by large, positive enthalpic and entropic changes. In contrast, the inactivation process observed for phm-MDH at low temperatures in the presence of various cosolvents was marked by a large, negative entropic contribution and a small, positive enthalpic contribution. The results obtained in this study indicate that more than one mechanism of inactivation can occur with these two multimeric enzymes depending on the selected temperature range and the type of cosolvent. The relationship of these results to stabilisation models for phm-MDH and ml-PEPC in the presence of various cosolvents, as well as the application of Arrhenius plot data to extrapolate the long term solution stability of these enzymes at lower temperatures from the pseudo-first order rate constants of inactivation experimentally derived over a range of temperatures, are discussed.

Stability studies on pig heart mitochondrial malate dehydrogenase: the effect of salts and amino acids.

The effect of different salts and amino acids on the thermal stability and quaternary conformation of pig heart mitochondrial malate dehydrogenase (phm-MDH) in solution has been determined. The effectiveness of salts of anions in the stabilisation of phm-MDH followed the order: Citrate > SO(4)2- > or = Tartrate > Phosphate > F-, CH3COO- > Cl- > Br-. Anions above and including Cl- in this series were increasingly effective in stabilising phm-MDH with a rise in salt concentration from 0.05-2 M, whilst Br- was destabilising under similar conditions. The effect of potassium salts of acetate, chloride and bromide at a concentration of 1 M on the quaternary conformation of phm-MDH correlated also with the relative order of anion stabilisation above, with the anions higher in the series increasingly promoting the formation of the dimeric conformation of the enzyme. The cations of the corresponding salts had a relatively neutral (Cs+, K+, Na+, (CH3)4N+, NH4+) to a destabilising ((CH3)4N+, NH4+, Li+) effect on phm-MDH. Potassium ferrocyanide and potassium ferricyanide conferred complex, concentration dependent effects on the stability of phm-MDH, unlike the salts described above. Salts of amino acids were effective in the stabilisation of phm-MDH against temperature induced changes, following the order: NaGlutamatec = NaAspartate > NaGlycinate > lysine. HCl > arginine. HCl. The magnitudes and trends of the effects of these salts and amino acids on the stability and quaternary structure of phm-MDH were observed to correlate well with considerations based on the Hofmeister series of anions and solvophobic concepts as they apply to the influence of co-solvents at intermediate to higher concentrations. Other, more specific effects were also evident in the stabilisation and destabilisation of phm-MDH by low concentrations of the salts, as noted most particularly in the presence of potassium ferrocyanide and potassium ferricyanide.

Hospital care by hospital-based and clinic-based faculty: a prospective, controlled trial.

BACKGROUND: The hospital length of stay decreases and clinical outcomes are maintained when teaching hospitals involve hospital-based attending physicians in comparison with traditional attending physicians. The attending physician's time commitment, including the number of hours per day and months per year, required to achieve this result is unknown. This study compared the clinical outcomes and cost of care for patients treated by hospital-based and clinic-based attending physicians devoting dramatically different amounts of time to supervising residents on the medical wards of a suburban county hospital. METHODS: Patients were alternately admitted to 2 groups of ward teams. Faculty who attended 10 months of the year supervised one group. The comparison group's attending physicians were on service for 2 months or less and maintained clinic responsibilities while on service. The cost of patient care was compared by means of the length of stay, total hospital costs, and costs for ancillary services. Hospital mortality and readmission rates compared clinical outcomes. RESULTS: There were 4456 patients hospitalized on the medical wards of a teaching service. No differences were detected in the length of stay (4.37 +/- 0.1 days for hospital-based and 4.39 +/- 0.1 days for clinic-based attending physicians). Hospital cost was observed to be similar (average cost, $5989 and $5977 per patient, respectively). The clinical outcomes were equivalent, with adjusted mortality rates for hospital-based attending physicians of 3.2% vs 3.9% for clinic-based attending physicians (P =.28). CONCLUSION: An increase of faculty time and involvement for supervision of resident-managed hospital care did not improve clinical outcomes or decrease costs during the 1-year study period.

Pulmonary complications of orthotopic liver transplantation.

Pulmonary complications following orthotopic liver transplantation (OLT) were prospectively evaluated in 18 individuals transplanted at the New England Deaconess Hospital. Of sixteen patients who survived the immediate postoperative period, 12 (75%) sustained a pulmonary complication. Of these complications, 64% were noninfectious--whereas 22% were infectious, and 14% probably infectious. Six of eight documented infections were caused by viruses of the herpes group. In four cases of viral pneumonitis other pulmonary pathogens were isolated (fungi-3, protozoan-1, bacteria-1). Unlike noninfectious complications, pulmonary infections were associated with a fatal outcome in five of six patients who died after OLT. Pulmonary complications are frequent and serious occurrences after OLT, and contribute to both the morbidity and mortality of this procedure. Compared with pulmonary complications seen after transplantation of other organs, OLT was associated with a higher proportion of noninfectious complications but a similar spectrum of pulmonary infections.

Myasthenia gravis presenting as isolated respiratory failure.

A patient with myasthenia gravis presenting as respiratory failure was unusual in his lack of peripheral neuromuscular involvement, negative results on many commonly used diagnostic tests, and lack of response to firstline therapeutic measures. Review of the pertinent literature revealed no previously described presentation of myasthenia gravis in this manner.

Abnormal airway function in individuals with the acquired immunodeficiency syndrome.

Pulmonary function test results of individuals with the acquired immunodeficiency syndrome (AIDS) were analyzed to determine the prevalence of abnormally low forced expiratory flow rates and bronchial hyperreactivity. Of 99 individuals with AIDS, a total of 44 (44 percent) had either low forced expiratory flow rates or a significant response to inhaled bronchodilator. Thirty-one (31 percent) individuals exhibited significant improvement in airflow rates after bronchodilator inhalation, while 33 (33 percent) had low flow rates. Twenty (20 percent) individuals had both low flow rates and a significant response to bronchodilator. In 83 percent of symptomatic individuals treated with bronchodilators there was clinical improvement. Thus, we conclude that abnormally low forced expiratory flow rate with or without bronchial hyperreactivity is a common and treatable complication of AIDS.

Analysis of induced sputum for the diagnosis of recurrent Pneumocystis carinii pneumonia.

We studied the sensitivity of ISA for diagnosis of second-episode PCP in AIDS patients. We induced sputum in 218 patients who had known or suspected AIDS and who had a presentation suggestive of PCP. All patients with negative sputum smear for PCP underwent BAL. Twenty-five patients were identified who had second-episode PCP at least 30 days after initial diagnosis. Chest roentgenographic infiltrate patterns for these 25 patients were blindly scored as normal, diffuse, upper lobe or focal non-upper lobe. The sensitivity of ISA was 72 percent for the first episode of PCP, 72 percent for the second episode of PCP, 72 percent for patients with second-episode PCP who had initial PCP detected by ISA and 71 percent for patients with second-episode PCP whose first episode of PCP was missed by ISA. Of the ten patients who were treated with AP, only one had a false-negative sputum analysis. A comparison of patients who had second-episode PCP diagnosed by ISA with those who had false-negative sputum analysis showed no difference in time to relapse, chest x-ray film pattern (all diffuse) or use of AP.

Controlled utilization of induced sputum analysis in the diagnosis of Pneumocystis carinii pneumonia.

We investigated the outcome and potential cost savings of a system designed to limit physician access to induced sputum analysis (ISA) for the diagnosis of Pneumocystis carinii pneumonia (PCP). Four respiratory medicine physicians screened all requests for ISA to determine the pretest likelihood of PCP. Twenty-two of 102 requests for ISA over a 1-year period were denied due to a low clinical suspicion for PCP. Seven individuals had a definitive alternative diagnosis confirmed and the remaining 15 were empirically treated for a presumptive diagnosis. All individuals were followed for at least 60 days or until death. None of the 22 individuals developed PCP during the follow-up period. We estimate that this approach saved $27,474, avoided exposure of health care workers to Mycobacterium tuberculosis, and was educational for the referring physicians.

Pneumoparotid due to spirometry.

Pneumoparotid has been described in patients who generate increased intraoral pressures when playing wind instruments, while coughing, and when undergoing dental work. Some patients have intentionally created pneumoparotid to avoid duties at school or in the military, or to gain attention. We describe a patient who developed pneumoparotid during pulmonary function testing. The diagnosis of pneumoparotid depends on a suggestive clinical situation and glandular swelling with or without crepitus. Observation of aerated saliva per Stensen's duct or air in the parotid duct and/or gland by any imaging study is diagnostic if infection with a gas-forming organism can be reasonably excluded. No specific treatment is required, other than the avoidance of predisposing activities.

Pulmonary mucormycosis presenting as an endobronchial lesion.

A 56-year-old diabetic man presented with left upper lobe collapse and postobstructive pneumonitis. Fiberoptic bronchoscopy revealed an endobronchial mass obstructing the left mainstem bronchus. The lesion resembled a bronchial adenoma; however, cytologic and histologic examination revealed invasive mucormycosis. The patient was treated with intravenous amphotericin B followed by endoscopic laser surgery that relieved the obstruction.

A modified Abrams needle biopsy technique.

STUDY OBJECTIVE: To compare the diagnostic sensitivity of a modified Abrams needle pleural biopsy technique (A1) with the standard Abrams (A2) and Cope needle biopsy methods. The modified Abrams pleural biopsy technique consisted of suctioning each tissue sample into a syringe without removing the needle completely from the chest until the completion of the entire procedure. Both the standard Abrams and Cope needle techniques required needle removal from the chest after each pleural biopsy. DESIGN: Retrospective chart analysis. SETTING: Community teaching hospital affiliated with Stanford University. PATIENTS: Forty-seven patients (30 men and 17 women) with a mean age 44.5 years (range, 19 to 81 years) who were referred to a pulmonary consultation service for pleural biopsy. INTERVENTIONS: Two of us (C. M. K. and F. T. K.) used the modified Abrams technique and two of us (W. A. J. and A. C. C.) used the standard Abrams technique. The Cope needle was used as originally described. MEASUREMENTS: We recorded the type of pleural biopsy needle and technique used in each patient. Biopsy specimen diameter and number of tissue samples obtained, final diagnoses, and complications were recorded. RESULTS: The diagnostic sensitivity for tuberculous pleurisy was 82% for the modified Abrams method, 71% for the standard Abrams method, and 88% for the standard Cope technique (p > or = 0.3). There was no difference in size of tissue sample obtained (A1 vs A2), number of biopsies, or complications among the three methods of pleural biopsy. CONCLUSIONS: The modified method of Abrams needle biopsy demonstrates a diagnostic sensitivity for pleural tuberculosis (82%) that is equivalent to that for the standard Abrams or Cope methods.

The prevalence and response to therapy of Strongyloides stercoralis in patients with asthma from endemic areas.

STUDY OBJECTIVE: To evaluate the prevalence and response to therapy of Strongyloides stercoralis infection in immigrant patients with asthma from areas endemic for Strongyloides. DESIGN AND INTERVENTIONS: In all patients, we performed a complete history and physical examination, complete blood cell counts (CBC), S stercoralis serologic tests, spirometry, and evaluated three stool samples for ova and parasites. Patients treated for S stercoralis infection had follow-up CBC, spirometry, serologic tests, and at least three additional stool examinations to confirm eradication of the parasite. SETTING: Ambulatory and hospitalized patients who were referred to the respiratory medicine clinic of a general hospital for the evaluation and treatment of asthma. PATIENTS: Forty-five asthmatic adults, representing 12 endemic countries, ranging in age from 20 to 76 years, were prospectively evaluated. RESULTS: Six of 45 patients were infected with S stercoralis, which yielded a prevalence of 13 percent. The patients with asthma and S stercoralis infection had higher blood eosinophil counts (p = 0.006) and were younger (p = 0.006) compared with patients with only asthma. There was no difference in the duration of asthma, spirometry, or steroid use between the two groups. Patients with S stercoralis and asthma tended to be more recent immigrants (p = 0.05). Five of the six patients with S stercoralis agreed to be treated with thiabendazole but only four returned for follow-up evaluation. All four patients had eradication of S stercoralis infection confirmed by negative stool examinations and a decline in S stercoralis serology (160 +/- 25 percent vs 13 +/- 13 percent, p = 0.03). All four patients had a decline in total blood eosinophil counts (2,476 +/- 832 cells per cubic millimeter vs 551 +/- 138 cells per cubic millimeter, p = 0.03) without a clinical improvement in asthma. CONCLUSIONS: Our data suggest that patients with asthma from areas endemic for S stercoralis, who have elevated peripheral blood eosinophil counts, should be screened for S stercoralis infection. Successful eradication of S stercoralis, however, may not result in a clinical improvement of asthma.

Bronchoscopic and radiologic features of Kaposi's sarcoma involving the respiratory system.

In a group of 61 patients admitted to New England Deaconess Hospital, Boston, with a diagnosis of acquired immune deficiency syndrome (AIDS), 25 were found to have Kaposi's sarcoma involving the skin and mucous membranes. Of these 25 patients, eight had lesions involving the respiratory system. Radiographically, patients with Kaposi's sarcoma had hilar and mediastinal adenopathy with perihilar parenchymal infiltration which progressed to diffuse bilateral infiltrates over a period of months. This pattern and the tempo of its evolution were distinctly different from the diffuse infiltrates seen in patients with Pneumocystis carinii pneumonia. Bronchoscopy was performed in seven of the eight patients, revealing characteristic lesions of Kaposi's sarcoma in the airways. We propose that parenchymal pulmonary Kaposi's sarcoma can be strongly suspected in a patient with AIDS who has the following features: a characteristic radiologic pattern; endobronchial Kaposi's sarcoma at bronchoscopy; and no evidence of opportunistic infection. In this subset of patients, further diagnostic intervention such as open lung biopsy, a procedure with potential morbidity in these ill individuals, may be unnecessary.

The optimal number of pleural biopsy specimens for a diagnosis of tuberculous pleurisy.

STUDY OBJECTIVES: To determine the optimal number of pleural biopsy (PLBX) specimens for a diagnosis of tuberculous pleurisy. DESIGN: Retrospective review. SETTING: County hospital. METHODS: We reviewed all percutaneous needle biopsy specimens of the parietal pleura in 30 patients who had tuberculous pleurisy. Data are reported as mean+/-SEM and statistical comparisons are done with the Mann-Whitney test. We accepted p<0.05 as statistically significant. RESULTS: The number of biopsy specimens obtained from each patient ranged from 4 to 10 with 1 sample submitted for mycobacterial culture and the rest submitted for histologic study. Sixty percent of patients had pleural cultures positive for Mycobacterium tuberculosis and 80% had diagnostic histology. Overall pleural biopsy sensitivity (histology and culture) for tuberculous pleurisy was 87%. On average, 40.4%+/-4.7% of all PLBX specimens contained pleura. Diagnostic PLBX procedures compared to false-negative procedures produced more tissue specimens (7.1+/-0.3 vs 4.8+/-0.5, p=0.005) containing more pleural specimens (2.4+/-0.2 vs 0.8+/-0.5, p=0.01). If only PLBX procedures yielding more than six tissue specimens (n=18) or more than two pleural specimens (n=12) were analyzed, then the diagnostic sensitivity of PLBX for pleural tuberculosis was 100%. There seemed to be a direct relationship between the sensitivity of PLBX and the number of specimens submitted. CONCLUSIONS: The sensitivity of percutaneous needle biopsy for diagnosis of tuberculous pleurisy is highest when more than six specimens are obtained which, on average, contain more than two specimens of parietal pleura. There are no conclusive data indicating how many tissue specimens to submit for mycobacterial culture, but one specimen seems sufficient.

Viral infection of the lower respiratory tract.

This article describes the epidemiology, transmission, and pathophysiology of viral infection of the lower respiratory tract. Current approaches to diagnosis, treatment, and prevention also are discussed.

Seroprevalence of bovine immunodeficiency-like virus and bovine leukemia virus in a dairy cattle herd.

To determine the prevalence of single vs. dual infection with bovine immunodeficiency virus (BIV) and bovine leukemia virus (BLV), sera (n = 95) from a dairy cattle herd were analyzed for anti-BIV and anti-BLV antibodies by an enzyme linked immunosorbent assay. Twenty-one percent (20/95) of samples were BIV-seropositive, while 52% (49/95) of the same samples were BLV-seropositive. A significantly greater percentage of BIV-seronegative samples were BLV-seropositive, 57% (43/75), than were BIV-seropositive samples, 30% (6/20). There was no significant correlation between data ranked from least to greatest amount of anti-viral antibody. Five cattle had persistent lymphocytosis (PL); all five were BLV-seropositive and two were BIV-positive. The mean anti-BLV titer was significantly greater in PL cattle, as compared at non-PL cattle, whereas there was no significant difference between the mean anti-BIV titer in PL cattle, as compared with non-PL cattle. These results provide additional information on the seroprevalence of naturally occurring BIV infection, and indicate that BIV can exist independent of other common infectious agents, such as BLV. Further, the results suggest that infection with BIV is not associated with an increased rate of infection with other infectious agents such as BLV.

In vitro expression of bovine leukemia virus in isolated B-lymphocytes of cattle and sheep.

The purpose of this study was to determine the effect of T-lymphocytes and phytohemagglutinin (PHA), a T-cell mitogen, on the expression of bovine leukemia virus (BLV) in cultured B-lymphocytes from BLV-infected cattle and sheep. Bovine B-lymphocytes were isolated by negative selection via complement-mediated lysis of T-lymphocytes. Additionally, bovine and ovine B-lymphocytes were positively selected using fluorescence activated cell sorting. Expression of BLV in cultured bovine and ovine B-lymphocytes occurred in the absence of T-lymphocytes and without PHA stimulation. The results of this study demonstrate that BLV replication in cultured B-lymphocytes is T-cell independent. This finding may have implications for the mechanism of viral latency within infected B-lymphocytes.