Mycobacterium interjectum Lung Infection.

A 62-year-old male presented with productive cough, weight loss, and night sweats. CXR revealed a right upper lobe cavitary lesion. Evaluation was negative for Mycobacterium tuberculosis, and sputum revealed Mycobacterium avium intracellulare (MAI). Since his clinical course was atypical for MAI, further investigations were pursued which identified Mycobacterium interjectum in lung specimens, a very rarely described etiology of pulmonary disease. Appropriate therapy with rifampin, intravenous amikacin, trimethoprim/sulfamethoxazole (TMP/SMX), and ethambutol resulted in clinical and radiographic improvement. This is the third case described over a period of 20 years of destructive lung disease in an immunocompetent adult due to M. interjectum.

Use of electron microscopy in core biopsy diagnosis of oncocytic renal tumors.

The distinction between oncocytoma and chromophobe renal cell carcinoma, important clinically, may be challenging, especially as the tissue sample size decreases. Ancillary studies can be helpful, although subject to interpretation and sample variability. The aim of this study was to examine the value of electron microscopy in differentiating between oncocytoma and chromophobe renal cell carcinoma on formalin fixed paraffin embedded needle core biopsies. Twenty renal needle core biopsies were evaluated. Despite formalin fixation and paraffin embedding, the classic ultrastructural features of these neoplasms were retained, revealing 80% sensitivity and 100% specificity by initial work-up.

Bordetella bronchiseptica pneumonia in a kidney-pancreas transplant patient after exposure to recently vaccinated dogs.

Bordetella bronchiseptica is an uncommon cause of respiratory infection in humans generally occurring in immunocompromised individuals exposed to infected animals. A 61-year-old female underwent a kidney-pancreas transplant 7 years before presentation. Postoperative immunosuppression was achieved with sirolimus and mycophenolate mofetil. The patient was doing well until she developed a small bowel obstruction secondary to adhesions. She underwent surgical adhesiolysis without complications. Two weeks postoperatively the patient developed pneumonia. She failed to respond to repeated courses of antibiotics, and thus, diagnostic bronchoscopy with bronchoalveolar lavage (BAL) was performed. BAL cultures grew B. bronchiseptica. Further investigation revealed that the patient's dogs had recently received a live-attenuated B. bronchiseptica intranasal vaccination. The patient recovered after 21 days of therapy with doxycycline based upon in vitro susceptibility testing. B. bronchiseptica is an uncommon but recognized human pathogen. It most commonly affects immunocompromised individuals. Here, we report a culture-proven case of B. bronchiseptica pneumonia in an immunocompromised host residing in a household with her dogs who had recently received live-attenuated, intranasal B. bronchiseptica vaccinations. As polymerase chain reaction testing was not performed comparing the isolated strain to the vaccination strain, the association is presumptive. This report expands the spectrum of immunocompromised hosts to include renal-pancreas transplant patients who have developed infection from B. bronchiseptica, while illustrating the risks associated with animal contacts and attenuated vaccines in the immunosuppressed population.

Phase II trial of imatinib mesylate in patients with metastatic melanoma.

Metastatic melanoma cells express a number of protein tyrosine kinases (PTKs) that are considered to be targets for imatinib. We conducted a phase II trial of imatinib in patients with metastatic melanoma expressing at least one of these PTKs. Twenty-one patients whose tumours expressed at least one PTK (c-kit, platelet-derived growth factor receptors, c-abl, or abl-related gene) were treated with 400 mg of imatinib twice daily. One patient with metastatic acral lentiginous melanoma, containing the highest c-kit expression among all patients, had dramatic improvement on positron emission tomographic scan at 6 weeks and had a partial response lasting 12.8 months. The responder had a substantial increase in tumour and endothelial cell apoptosis at 2 weeks of treatment. Imatinib was fairly well tolerated: no patient required treatment discontinuation because of toxicity. Fatigue and oedema were the only grade 3 or 4 toxicities that occurred in more than 10% of the patients. Imatinib at the studied dose had minimal clinical efficacy as a single-agent therapy for metastatic melanoma. However, based on the characteristics of the responding tumour in our study, clinical activity of imatinib, specifically in patients with melanoma with certain c-kit aberrations, should be examined.

Persistence of Campylobacter species, strain types, antibiotic resistance and mechanisms of tetracycline resistance in poultry flocks treated with chlortetracycline.

OBJECTIVES: The aim of this study was to investigate the persistence of Campylobacter species, strain types, antibiotic resistance and mechanisms of tetracycline resistance in poultry flocks treated with chlortetracycline. METHODS: Three commercially reared broiler flocks, naturally colonized with Campylobacter, were treated with chlortetracycline under experimental conditions. The numbers of Campylobacter isolated, and the species, flaA short variable region allele, and antimicrobial resistance of isolates were determined. RESULTS: For two of three flocks, tetracycline-resistant strains predominated prior to chlortetracycline exposure. Presence of the antibiotic had no discernible effect on the numbers or types of Campylobacter and the tetracycline-resistant strains persisted in numbers similar to those observed before treatment. With all flocks, some faecal samples were obtained that contained no Campylobacter, irrespective of exposure to chlortetracycline; this was more common as the birds grew older. For the third flock, tetracycline-resistant Campylobacter were in the minority of samples before and during exposure to chlortetracycline, but at sampling times after this, no resistant strains were found in the treated (or untreated) birds, irrespective of exposure to the antibiotic. All tetracycline-resistant isolates (MICs 16 to >128 mg/L) contained tet(O) and, for some isolates, this was transferable to Campylobacter jejuni 81116. The efflux pump inhibitor PAbetaN reduced the MICs of tetracycline for these isolates by 4-fold, suggesting that an intact efflux pump, presumably CmeABC, is required for high-level tetracycline resistance. CONCLUSIONS: Our data indicate that chlortetracycline treatment does not eradicate tetracycline-resistant Campylobacter spp. from poultry. However, if a low number of resistant isolates are present, then the antibiotic pressure appears insufficient to select such strains as the dominant population.

Regulation of iNOS by the p44/42 mitogen-activated protein kinase pathway in human melanoma.

Activating mutations of the genes for NRAS and BRAF, components of the p44/42 mitogen-activated protein kinase (MAPK) pathway, are common findings in melanoma. Recent evidence in several nonmelanoma cell systems supports the regulation of the inducible nitric oxide synthase (iNOS) gene by this pathway. On the basis of our data showing that melanoma iNOS expression predicts shortened patient survival, we formulated the hypothesis that activating mutations of NRAS or BRAF, which lead to constitutive activation of the p44/42 MAPK pathway, drive iNOS expression in human melanoma. In the present study, we have shown that inhibition of melanoma iNOS activity by S-methylisothiourea leads to decreased cell proliferation, confirming the importance of iNOS activity for melanoma cell growth. Regulation of melanoma iNOS expression by the p44/42 MAPK pathway was demonstrated by inhibition of the pathway by U0126, and by BRAF RNA interference. To explore this regulatory pathway in human tissue, 20 melanoma tumors were examined for NRAS and BRAF mutations, immunohistochemical evidence of ERK phosphorylation, and iNOS expression. A significant association was found among these three features. We conclude that in human melanoma, activating mutations of NRAS and BRAF drive constitutive iNOS expression and, implicitly, nitric oxide production, contributing to the poor survival of these patients.

Development of a standardized susceptibility test for campylobacter with quality-control ranges for ciprofloxacin, doxycycline, erythromycin, gentamicin, and meropenem.

A standardized agar dilution susceptibility testing method was developed for Campylobacter that consisted of testing on Mueller-Hinton medium supplemented with 5% defibrinated sheep blood in an atmosphere of 10% CO2, 5% O2, and 85% N2. Campylobacter jejuni ATCC 33560 was identified as a quality-control (QC) strain. Minimal inhibitory concentration (MIC) QC ranges were determined for two incubation time/temperature combinations: 36 degrees C for 48 hr and 42 degrees C for 24 hr. Quality-control ranges were determined for ciprofloxacin, doxycycline, erythromycin, gentamicin, and meropenem. For all antimicrobial agents tested at both temperatures, 95-100% of the QC MIC results fell within recommended QC ranges. Twenty-one Campylobacter clinical isolates, encompassing five species of Campylobacter (C. jejuni, C. coli, C. jejuni, subsp. doylei, C. fetus, and C. lari) were tested in conjunction with the C. jejuni QC strain. While C. jejuni and C. coli could be reliably tested under both test conditions, growth of C. jejuni subsp. doylei, C. fetus, and C. lari isolates was inconsistent when incubated at 42 degrees C. Therefore, it is recommended that these species only be tested at 36 degrees C.

Activity of faropenem and imipenem for ciprofloxacin-resistant bacteria.

AIM: To determine whether an association exists between ciprofloxacin and faropenem resistance in bacteria including multiply drug-resistant isolates. METHODS: The MICs were determined for 150 fluoroquinolone-resistant bacteria, plus 20 nalidixic acid-resistant strains of Salmonella enterica serovar Typhimurium. RESULTS: Faropenem was very active against Escherichia coli and Streptococcus pneumoniae, but 5/31 Staphylococcus aureus and 2/26 Bacteroides fragilis required > or =16 mg/L for inhibition. Of 30 multiply drug-resistant isolates with a phenotype suggestive of enhanced efflux, only for one strain (a Bacteroides fragilis) was the faropenem MIC higher than that associated with the other isolates of the same species. CONCLUSIONS: Faropenem was in general as active as imipenem. There was no association between resistance to ciprofloxacin and faropenem or imipenem resistance.

Accumulation of 10 fluoroquinolones by wild-type or efflux mutant Streptococcus pneumoniae.

A method for measuring fluoroquinolone accumulation by Streptococcus pneumoniae was rigorously examined. The accumulation of ciprofloxacin, clinafloxacin, gatifloxacin, grepafloxacin, levofloxacin, moxifloxacin, norfloxacin, ofloxacin, sitafloxacin, and trovafloxacin in the presence and absence of either carbonyl cyanide m-chlorophenyl-hydrazone (CCCP) or reserpine was determined for two wild-type fluoroquinolone-susceptible capsulated S. pneumoniae strains (M3 and M4) and the noncapsulated strain R6. Two efflux mutants, R6N (which overexpresses PmrA) and a mutant of M4, M22 (no expression of PmrA), were also examined. Essentially, the fluoroquinolones fell into two groups. (i) One group consisting of ciprofloxacin, grepafloxacin, and norfloxacin accumulated to 72 to 92 ng/mg (dry weight) of cells in all strains. (ii) The remainder of the agents accumulated to 3 to 30 ng/mg (dry weight) of cells. With a decrease in hydrophobicity, there was a decrease in the concentration accumulated. With an increase in the molecular weight of the free form of each agent, there was also a decrease in the concentration accumulated. The strains differed in their responses to reserpine and CCCP. For the three fluoroquinolone-susceptible strains, only reserpine had a significant effect upon accumulation of moxifloxacin and clinafloxacin by M3 and showed no effect for the other agents and strains. For M3 and M4, CCCP enhanced the concentration of ciprofloxacin and norfloxacin accumulated, whereas for R6, the effect was only statistically significant for ofloxacin. Efflux mutant M22 accumulated less ciprofloxacin, gatifloxacin, and ofloxacin than M4 did. M22 accumulated more norfloxacin than M4 did. Reserpine and CCCP had variable effects as for the other strains. Differences in the accumulation of fluoroquinolones by R6 and R6N were highly dependent upon growth phase, and only for norfloxacin was there a significant difference between two strains.

An optimized system for studies of EPO-dependent murine pro-erythroblast development.

OBJECTIVE: Objectives were to develop new means to isolate useful numbers of primary progenitor cells and to quantitatively assay the stepwise maturation of erythroblasts. METHODS: Approaches involved dosing mice with thiamphenicol (TAP) to yield staged cohorts of pro-erythroid cells; optimizing conditions for their EPO-dependent in vitro growth and survival; developing assays for CFU-E maturation; analyzing stage-specific transcript expression; and expressing a heterologous, erythroid-specific tag (EE372) in transgenic mice. RESULTS: Per TAP-treated mouse, 3 x 10(7) highly EPO-responsive erythroid progenitor cells were generated that represented up to 30% of total splenocytes and showed strict dependence on EPO for survival, growth, and immediate response gene expression. In this developing cohort, a tightly programmed sequence of gene expression was observed, and maximal expression of c-kit, EPO receptor, and beta-globin transcripts occurred at 72, 96, and 120 hours post-TAP withdrawal, respectively. Also, the newly discovered erythroid-specific dual-specificity kinase, DYRK3, was revealed to be expressed at a late CFU-E stage. In vitro, these progenitor cells matured stepwise from high FALS Ter119- cells (24-hour culture) to high FALS Ter119+ cells (24-36 hours) to low FALS Ter119+ maturing erythroblasts (40-48 hours) and sharp differences in their morphologies were observed. Finally, a MACS-based procedure for the purification of erythroid progenitor cells from TAP-treated EE372 transgenic mice also was developed. CONCLUSIONS: A comprehensive new system for isolating large numbers of primary murine erythroid progenitor cells and quantitatively monitoring their development is established that should serve well in investigations of endogenous and pharmacological regulators of red blood cell development.

Urban-rural differences in a memory disorders clinical population.

OBJECTIVES: To compare patient characteristics and family perceptions of patient function at one urban and one rural memory disorders clinic. DESIGN: Secondary, cross-sectional data analyses of an extant clinical database. SETTING/PARTICIPANTS: First time visits (n = 956) at two memory disorders clinics. MEASUREMENTS: Patient and family-member demographics and assessment results for the Mini-Mental State Examination (MMSE), instrumental activities of daily living (IADLs), activities of daily living (ADLs), the Memory Change and Personality Change components of the Blessed Dementia Rating Scale, and the Revised Memory and Behavior Problems Checklist. RESULTS: In both clinics, patients and family members were more likely female. The typical urban clinic patient was significantly more likely to be living in a facility and more educated than the typical rural patient. Urban and rural patients did not show significant differences in age- and education-adjusted MMSE scores or raw ADL/IADL ratings, but the urban family members reported more memory problems, twice as many personality changes, more-frequent behavior problems, and more adverse reactions to problems. CONCLUSION: Physicians who practice in both urban and rural areas can anticipate differences between patients, and their families, who seek a diagnosis of memory disorders. Our most important finding is that despite similarities in reported functional abilities, urban families appear to be more sensitive to and more distressed by patients' cognitive and behavioral symptoms than rural families. These differences may reflect different underlying needs, and should be explored in further research.

Effective antigen-specific immunotherapy in the marmoset model of multiple sclerosis.

Mature T cells initially respond to Ag by activation and expansion, but high and repeated doses of Ag cause programmed cell death and can suppress T cell-mediated diseases in rodents. We evaluated repeated systemic Ag administration in a marmoset model of experimental allergic encephalomyelitis that closely resembles the human disease multiple sclerosis. We found that treatment with MP4, a chimeric, recombinant polypeptide containing human myelin basic protein and human proteolipid protein epitopes, prevented clinical symptoms and did not exacerbate disease. CNS lesions were also reduced as assessed in vivo by magnetic resonance imaging. Thus, specific Ag-directed therapy can be effective and nontoxic in primates.

Diversity in medical school: perceptions of first-year students at four southeastern U.S. medical schools.

PURPOSE: To assess students' perceptions of the extent of diversity in their classes, the role of diversity in their first-year curriculum, and their predictions of the amount of diversity in their future patient populations. METHOD: In 1998, students at four southeastern U.S. medical schools that had distinct demographics and differing institutional missions completed a questionnaire on diversity at the end of the first year. In the instrument, diversity was defined according to nine population characteristics: age, sex, race, ethnic background, physical disability, religious affiliation, sexual orientation, socioeconomic status, and rural background (growing up in a community of less than 5,000). Responses were compared according to students' institution, sex, and race. RESULTS: Questionnaires were returned by 349 of 474 students (74%). Students at the school with the most diverse first-year class placed the greatest value on the contributions of diversity to the learning environment. Women students placed more value on the inclusion of diversity issues in the curriculum than did men students, and they placed greater value on understanding diversity issues in their future medical practices than did men. Compared with Asian American, Hispanic, and white students, African American students were the least likely to think that the curriculum contained adequate information about diversity. CONCLUSIONS: The results indicate that perceptions of diversity were influenced by the students' own demographic characteristics and those of their medical school. The more diverse the class, the more comfortable the students were with diversity and the more they valued its contribution to their medical education.

Effect of external dead volume on performance while wearing a respirator.

Respirator dead volume accumulates exhaled carbon dioxide and returns it to the respiratory system during subsequent inhalations. Because inhaled carbon dioxide is known to be a powerful respiratory stimulant and psychoactive gas, respirator dead volume would be expected to influence performance times while respirators are worn during work. This experiment was performed at intense levels of treadmill walking (80-85% VO2max) to demonstrate maximum sensitivity to respiratory stress. Six dead volume conditions (representing a range of 280 to 1,160 mL) were imposed on the subjects. Results show linear decreases in performance times and breathing apparatus comfort with increasing dead volumes. For each 350 mL of external dead volume, a 19% decrease of performance time and an 18% decrease in breathing apparatus comfort can be expected.

Purchasing over-the-counter medications: the influence of age and familiarity.

This study examines the influence of age and type of over-the-counter (OTC) medications on decision-making processes leading to the selection of OTC medications. Manipulating type of OTC medication served as a way of examining subjects' familiarity with the decision domain. Thirty-six younger and thirty-six older adults answered questions regarding their OTC medication purchases, and completed a decision task in which they searched four computerized displays of product label information in order to select an antacid, a cold medication, a laxative, and a pain reliever. In general, older adults were slower to review information, more likely to have used OTC medications, and more organized in their searches for information. Specific to pair relievers, older and younger adults demonstrated similar information use and time to decision, and older adults specifically tailored the organization of their information searches when choosing pain relievers. Clearly, older adults selectively use product information on OTC medication packages, given adequate time to process information. Manufacturers' efforts directed at improving information availability should benefit older adults' decision processes.

Addition of eicosapentaenoic acid to gamma-linolenic acid-supplemented diets prevents serum arachidonic acid accumulation in humans.

Previous studies reveal that supplementation of human diets with gamma-linolenic acid (GLA) reduces the generation of lipid mediators of inflammation and attenuates clinical symptoms of chronic inflammatory disorders such as rheumatoid arthritis. However, we have shown that supplementation with this same fatty acid also causes a marked increase in serum arachidonate (AA) levels, a potentially harmful side effect. The objective of this study was to design a supplementation strategy that maintained the capacity of GLA to reduce lipid mediators without causing elevations in serum AA levels. Initial in vitro studies utilizing HEP-G2 liver cells revealed that addition of eicosapentaenoic acid (EPA) blocked Delta-5-desaturase activity, the terminal enzymatic step in AA synthesis. To test the in vivo effects of a GLA and EPA combination in humans, adult volunteers consuming controlled diets supplemented these diets with 3.0 g/d of GLA and EPA. This supplementation strategy significantly increased serum levels of EPA, but did not increase AA levels. EPA and the elongation product of GLA, dihomo-gamma-linolenic acid (DGLA) levels in neutrophil glycerolipids increased significantly during the 3-wk supplementation period. Neutrophils isolated from volunteers fed diets supplemented with GLA and EPA released similar quantities of AA, but synthesized significantly lower quantities of leukotrienes compared with their neutrophils before supplementation. This study revealed that a GLA and EPA supplement combination may be utilized to reduce the synthesis of proinflammatory AA metabolites, and importantly, not induce potentially harmful increases in serum AA levels.

Acquired thermotolerance and expression of the HSP100/ClpB genes of lima bean.

Acquired thermotolerance (AT) is the ability of cells to survive a normally lethal temperature treatment as a consequence of pretreatment at an elevated but sublethal temperature. In yeast and cyanobacteria, the expression of the HSP100/ClpB protein is required for the AT response. To determine whether the HSP100/ClpB protein is associated with this response in lima bean (Phaseolus lunatus), we have cloned an HSP100/ClpB homolog and assessed expression of the two gene copies under heat stress conditions, which induce AT. Transcription of the cytoplasmically localized HSP100/ClpB protein genes is stringently controlled by heat stress in both of the laboratory and field heat stress conditions. From a heat-induced cDNA library, we identified a clone of a putative chloroplast-targeted (cp) HSP100/ClpB protein gene sequence. The cp HSP100/ClpB protein genes are constitutively expressed, but transcript levels increase post-heat stress in laboratory heat stress experiments. In field conditions the genes for the cp HSP100/ClpB are constitutively expressed. Although we were unable to correlate differences in the timing of AT response with the expression or genetic structure of the HSP100/ClpB genes in heat-tolerant or -sensitive varieties of lima bean, we clearly demonstrate the association of expression of HSP100/ClpB proteins with heat response in this species.

Communication using a telephone while wearing a respirator.

Respirators have been found to degrade communication effectiveness when wearers speak face-to-face. However, little is known about communication effectiveness when using the telephone and wearing a respirator. Eleven pairs of subjects were asked to pronounce and identify words chosen from Modified Rhyme Test lists. Each word appeared on a computer screen in one room and the speaker said the word into the telephone. The listener in another room identified the word and typed it into a computer linked with the first. Subjects wore U.S. Army M40 full-facepiece air-purifying respirators with hoods. Three different speech diaphragm arrangements and two hood materials were tested. Results show that accuracy suffered by about 10% when respirators and hoods were worn compared with the control condition. Word identification speed was one-third to one-half of the control (no respirator or hood) condition depending on specific equipment worn.

Artifacts and the visualization of fetal distal extremities using three-dimensional ultrasound.

OBJECTIVES: To demonstrate that acoustic shadowing in 3D US may give rise to artifacts simulating limb defects and provide a solution to eliminate its occurrence. METHODS: Twenty second trimester fetuses (gestational age 15-24 weeks) were scanned with three-dimensional ultrasound (3D US) using a sagittal acquisition plane. Fetal tibia/fibula and radius/ulna pairs were assessed for completeness of imaging. A further 20 fetuses (gestational age 20-26 weeks) were scanned in both axial and sagittal planes and the results compared to verify clear visualization of both bones. RESULTS: Shadowing from adjacent structures produced an apparent limb defect in 55% of the first 20 fetuses imaged only sagittally (18% of limb pairs). Acquiring data from more than one orientation avoided this artifact. CONCLUSIONS: The 3D US is subject to the same artifacts as two-dimensional (2D US) in terms of acoustic shadowing, although their presentation may be different. Awareness of this fact is essential for correct interpretation of 3D US studies. Three-dimensional scanning protocols should be modified to ensure that fetal structures are adequately visualized by acquiring volume data in more than one acquisition orientation.