Pan-azole- and multi-fungicide-resistant Aspergillus fumigatus is widespread in the United States.

Aspergillus fumigatus is an important global fungal pathogen of humans. Azole drugs are among the most effective treatments for A. fumigatus infection. Azoles are also widely used in agriculture as fungicides against fungal pathogens of crops. Azole-resistant A. fumigatus has been increasing in Europe and Asia for two decades where clinical resistance is thought to be driven by agricultural use of azole fungicides. The most prevalent mechanisms of azole resistance in A. fumigatus are tandem repeats (TR) in the cyp51A promoter coupled with mutations in the coding region which result in resistance to multiple azole drugs (pan-azole resistance). Azole-resistant A. fumigatus has been isolated from patients in the United States (U.S.), but little is known about its environmental distribution. To better understand the distribution of azole-resistant A. fumigatus in the U.S., we collected isolates from agricultural sites in eight states and tested 202 isolates for sensitivity to azoles. We found azole-resistant A. fumigatus in agricultural environments in seven states showing that it is widespread in the U.S. We sequenced environmental isolates representing the range of U.S. sample sites and compared them with publicly available environmental worldwide isolates in phylogenetic, principal component, and ADMIXTURE analyses. We found worldwide isolates fell into three clades, and TR-based pan-azole resistance was largely in a single clade that was strongly associated with resistance to multiple agricultural fungicides. We also found high levels of gene flow indicating recombination between clades highlighting the potential for azole-resistance to continue spreading in the U.S.IMPORTANCEAspergillus fumigatus is a fungal pathogen of humans that causes over 250,000 invasive infections each year. It is found in soils, plant debris, and compost. Azoles are the first line of defense antifungal drugs against A. fumigatus. Azoles are also used as agricultural fungicides to combat other fungi that attack plants. Azole-resistant A. fumigatus has been a problem in Europe and Asia for 20 years and has recently been reported in patients in the United States (U.S.). Until this study, we did not know much about azole-resistant A. fumigatus in agricultural settings in the U.S. In this study, we isolated azole-resistant A. fumigatus from multiple states and compared it to isolates from around the world. We show that A. fumigatus which is resistant to azoles and to other strictly agricultural fungicides is widespread in the U.S.

Analysis of low-speed rotational atherectomy for the reduction of platelet aggregation.

Platelet activation is an important determinant of acute outcomes of percutaneous intervention. The objective of this study was to assess the effect of rotational atherectomy on platelet activation in an in vitro model. Freshly collected heparinized porcine blood was exposed to a 2.0-mm Rotablator burr rotating at one of three speeds: 180,000, 140,000, or 0 rpm. The specimens were analyzed immediately for concentration and size of platelet aggregates and plasma-free hemoglobin. There were significantly more platelet aggregates of >20-microm diameter at higher speeds (7,434+/-2,193 at 180,000, vs. 2,269+/-627 at 140,000, vs. 633+/-258 aggregates/ml at 0 rpm; P < 0.001). Plasma-free hemoglobin, a simple measure of cell damage, decreased with decreasing rotational speed (429+/-168 mg/dl at 180,000, vs. 88+/-44 mg/dl at 140,000, vs. 9+/-9 mg/dl at 0 rpm; P < 0.0001). In vitro, platelet activation decreases with decreasing burr speed, suggesting that the use of the Rotablator system at its minimum approved speed (140,000 rpm) could prove clinically beneficial.

Analysis of heat generation during rotational atherectomy using different operational techniques.

Rotational atherectomy can generate heat from the friction of the burr as it ablates atherosclerotic plaque. The objective of this study is to correlate Rotablator technique and heat generation using two experimental models. First, 2.0 mm burrs were advanced through a lesion model derived from bovine bone implanted with thermal probes. Intermittent ablation with minimal decelerations resulted in a smaller temperature increase than continuous ablation with maximal decelerations (2.6 +/- 1.3 vs. 13.9 +/- 1.0 degrees C, respectively, P < 0.01). The second model used porcine femoral arteries cradled in constricting polyethylene grafts with thermal probes in contact with the adventitia. As the burr advanced through the segment, RPM decreases of 5-7 k resulted in a temperature rise of 4.1 +/- 1.2 degrees C, whereas decelerations of 10-20 k resulted in a 11.3 +/- 6.2 degrees C temperature increase. We conclude that excessive drops in speed and aggressive advancement of the burr are related to significant increases in temperature and potential thermal injury.

Histological presence of viable prostatic glands on routine biopsy following cryosurgical ablation of the prostate.

PURPOSE: Cryosurgical ablation of the prostate has recently received much attention as a therapeutic alternative for the treatment of localized prostatic adenocarcinoma. Biopsies after treatment reveal a variety of dysplastic changes as well as unaltered prostatic glandular epithelial elements. Prostate specific antigen (PSA) remains undetectable in the majority of men. However, in some PSA increases without demonstrable local recurrence. MATERIALS AND METHODS: A total of 383 patients underwent 447 procedures between June 1990 and January 1994. Of 358 biopsies performed at our institution, 317 (2,075 cores) were available for review. Each core was examined for unaltered prostatic glandular epithelial elements and then scored for the percentage of epithelial glandular involvement according to a scale of: 0-no, 0.5-less than 10%, 1-10 to 25%, 2-25 to 50%, 3-50 to 75% and 4-76 to 100% unaltered prostatic glandular epithelial elements. RESULTS: Of 317 biopsies 158 (49.8%) contained no unaltered prostatic glandular epithelial elements, while 185 (58.3%) and 206 (65%) had 1 core containing 10% and 10 to 25%, respectively, of such elements. Of 262 cases (82.6%) with a mean of 10% unaltered prostatic glandular epithelial elements per core 22 (8.4%) were positive for residual carcinoma. Among 55 cases with more normal epithelium per core 24 (43.6%) were positive for residual carcinoma. Patients with a positive biopsy had a median PSA of 2.02 ng./ml. (average gland/core score 0.54). Median PSA for men with negative biopsies was 0.2 ng./ml. (gland/core score 0.124). CONCLUSIONS: Cryosurgical ablation of the prostate has the ability to ablate prostatic tissue completely, thus rendering it free of glandular elements as determined by biopsy. Increasing PSA can indicate residual glandular elements. Increases in unaltered prostatic glandular epithelial elements with time are not paralleled by increased rates of local disease recurrence. undetectable serum PSA has a low risk of residual unaltered prostatic glandular epithelial elements and localized carcinoma. Results as measured by unaltered prostatic glandular epithelial elements and PSA improve with the surgical experience.

Simultaneous radical nephrectomy and repair of abdominal aortic aneurysm.

OBJECTIVES: The combination of abdominal aortic aneurysm repair with other intra-abdominal surgery is controversial. Most studies have shown that a variety of procedures can be performed at the same time as an aneurysm repair with little change in mortality or complication rates. We conducted a retrospective study to determine if aneurysm repair could be safely and effectively combined with radical nephrectomy. METHODS: We studied 10 patients who underwent combined abdominal aortic aneurysm repair and radical nephrectomy during a 4-year period. Results from this group were compared to a separate control group of 10 patients who underwent radical nephrectomy alone and another of 12 patients underwent abdominal aortic aneurysm repair alone, during the same time period. RESULTS: The overall mortality was 10% and significant complications occurred in an additional 10% of patients. Minor, self-limiting complications occurred in 30% of patients. There were no aortic graft infections that occurred in the entire series of patients at 18 months of mean follow-up. There were no remarkable differences in the entire series of patients and the combined values in a separate group of control patients who had undergone either procedure alone. CONCLUSIONS: Simultaneous radical nephrectomy for presumed renal cell carcinoma can be safely combined with repair of abdominal aortic aneurysm in selected patients.

Percutaneous, transperineal cryosurgery of the prostate as salvage therapy for post radiation recurrence of adenocarcinoma.

BACKGROUND: Surgical techniques utilizing the application of very low temperatures to malignant tissues have been used increasingly in recent years in the minimally invasive treatment of prostate cancer. An area of potential application appears to be in the management of radiation resistant prostate cancer. METHODS: This study represents a retrospective chart review of 33 patients undergoing cryosurgical ablation of the prostate (CSAP) according to a protocol designed by an institutional review board at a single institution for the treatment of radiation resistant prostate cancer. Radiation resistance was defined as a positive prostate needle biopsy and rising prostate specific antigen (PSA). Treatment effect was assessed by serial post treatment PSA determinations and extensive systematic post treatment prostate needle biopsies. RESULTS: Of the 33 patients analyzed, 24 converted to an all negative biopsy status after one CSAP treatment. Repeat treatment converted two additional patients to a biopsy negative status. CSAP appeared to lower PSA dramatically in most patients. One year after treatment, of 10 patients not on androgen deprivation therapy, 3 maintained a PSA of < 0.4 ng/mL and those patients with androgen deprivation therapy maintained a PSA of < 4.0 ng/mL. The most frequent complications included sloughing syndrome (15.4%) and incontinence (10.3%). CONCLUSIONS: CSAP appears to eliminate biopsy detectable prostate tumor effectively in the majority of cases in the salvage setting following radiation therapy. A significant discrepancy exists, however, between the biopsy "cure" rate and the biochemical "cure" rate, even in the short term. There is a suggestion that early detection of radiation failure may improve results of this salvage therapy.

Cryosurgical ablation of the prostate: two-year prostate-specific antigen and biopsy results.

OBJECTIVES: Percutaneous cryosurgical ablation of the prostate (CSAP) was performed on patients with localized or locally advanced adenocarcinoma of the prostate. To assess local disease control, post-treatment biopsy and serum prostate-specific antigen (PSA) levels were obtained at 3 and 24 months post-treatment. METHODS: From June 1990 through May 1994, CSAP was performed 448 times on 383 patients under Institutional Review Board protocols. A urethral warming catheter was used for all procedures. A total of 239 patients were followed for a minimum of 21 months after treatment. None of this group had received prior local treatment. The group consisted of patients who were newly diagnosed and treated solely with cryotherapy (virgin); the remainder had been on androgen deprivation therapy (ADT) prior to CSAP. RESULTS: Biopsies were obtained from 114 patients at 21 months or more after treatment. In the virgin group, 79% had a negative biopsy after one or more treatments, and 88% of the ADT group are negative after one or more treatments. Overall, 69% had a negative biopsy after one treatment and 82% had a negative biopsy following one or more CSAP treatments. Of a group of 163 patients, PSA data were evaluable at 21 months or more after treatment. In the virgin group, 60% had a PSA 0.4 ng/mL or less, and 77% had a PSA 1.0 ng/mL or less. In the ADT group, 40% had a PSA 0.4 ng/mL or less, and 69% had a PSA value of 1.0 ng/m Lor less. Complications were minimal, the most common one being urethral tissue sloughing, which occurred in 10% of patients. CONCLUSIONS: CSAP appears to be effective in obtaining local control as measured by biopsy and PSA 21 months or more post-treatment. When retrospectively comparing our results with recently published radiotherapy series, CSAP was more effective in obtaining nadir PSA values 1.0 ng/mL or less and negative biopsies at 21 months or more after treatment.

Cryosurgical ablation of the prostate: a nursing perspective.

1. Cryosurgical ablation of the prostate is an attempt to use the proved cytodestructive abilities of cryosurgery in combination with the guidance of transrectal ultrasound to provide local disease control with less morbidity and a quicker recovery period. 2. Cryosurgical ablation of the prostate begins as a same day admission. Patients are out of bed and eating as tolerated that evening. They are generally discharged the following day if able to manage the suprapubic tube. 3. Though the recovery period is generally a quick one, there exist many idiosyncrasies to this recovery which make careful interaction between the physician, nurse, and patient critical.

Urethral warming catheter for use during cryoablation of the prostate.

Cryoablation of the prostate has been shown to be a useful tool in the eradication of prostate cancer. Sloughing of necrotic tissue per urethra was one of the major complications of prostate cryosurgery during its initial application and led to its disuse by urologists. A catheter was then needed until sloughing was complete and the patient again able to void. In a previous report, we have demonstrated that thermal protection of the urethra could be obtained by flowing warm irrigant through a thin-walled urethral conduit. To simplify the application of this heat, a dual lumen urethral catheter has been designed that can be passed transurethrally to circulate warm saline and prevent the destruction of the urethral epithelium. This prevents transmural necrosis, maintaining the epithelial barrier for containment of necrotic prostate tissue after treatment. This urethral warming device is made of materials commonly found in operative stockrooms. To date, the device has functioned well with a urethral sloughing rate of 12% during its initial use.

Cloning, nucleotide sequence, and expression of a p-hydroxybenzoate hydroxylase isozyme gene from Pseudomonas fluorescens.

A gene encoding for a putative isozyme of p-hydroxy-benzoate hydroxylase (PHBH) has been isolated from Pseudomonas fluorescens (ATCC 13525). A comparison of the translated amino acid sequence with that of the known PHBH from P. fluorescens revealed that the new enzyme contains 3 additional amino acids and has 73% absolute homology to the previously known enzyme; conservation of secondary and active-site structures implied that the isozyme and known enzyme share the same general tertiary structure. Subsequent expression of the isozyme in Escherichia coli produced an enzyme with a specific activity about half that of the previously characterized PHBHs from P. fluorescens and Pseudomonas aeruginosa; in addition, somewhat weaker binding affinities for both NADPH and p-hydroxybenzoate were observed. Speculations are made on the reason for the existence of the isozyme, which does not appear to be expressed routinely in P. fluorescens.

Stability of expression of Neisseria gonorrhoeae lipooligosaccharides.

We compared multiple lipooligosaccharide (LOS) extracts from individual strains of Neisseria gonorrhoeae. Each of the extracts was prepared from single mass cultures grown on solid media under similar conditions but separated by time. We found only subtle variations in the number, electrophoretic mobility, and concentration of components of the LOSs from individual strains. We found no variation in the expression of a 3.6-kilodalton LOS component that carries the L8 LOS epitope. A significant variation in the 3-deoxy-D-manno-octulosonic acid content was found among different extracts from the same strain, but this variation appeared to be unrelated to the other LOS characteristics studied.

Preventive intervention during the perinatal and infancy periods: overview and guidelines for evaluation.

This paper reviews recent changes in perspectives of development in the prenatal, neonatal, and infancy periods and describes exemplary preventive interventions and their evaluation. Particular emphasis is placed on expanding criteria for successful early interventions to include measures of socioemotional and physical health as well as the more traditional measures of intellectual development. A theme which emerges is the need for peer and professional support for early parenting.

A model for teaching early child development.

A model of teaching psychological aspects of infant growth and development to medical students in a pediatric clinical rotation is described. Small groups of 8 to 10 medical students observe an infant developmental-assessment-parent-interview conducted by a child development specialist. Emphasis is placed upon demonstrating infant temperament, age-appropriate competencies, and the dynamic interplay between parent and child. Evaluation of this training program suggests its effectiveness as a method of instruction and its usefulness as an approach to the evaluation of the young child.