Bacteria in a water-damaged building: associations of actinomycetes and non-tuberculous mycobacteria with respiratory health in occupants.

We examined microbial correlates of health outcomes in building occupants with a sarcoidosis cluster and excess asthma. We offered employees a questionnaire and pulmonary function testing and collected floor dust and liquid/sludge from drain tubing traps of heat pumps that were analyzed for various microbial agents. Forty-nine percent of participants reported any symptom reflecting possible granulomatous disease (shortness of breath on exertion, flu-like achiness, or fever and chills) weekly in the last 4 weeks. In multivariate regressions, thermophilic actinomycetes (median = 529 CFU/m2 ) in dust were associated with FEV1 /FVC [coefficient = -2.8 per interquartile range change, P = 0.02], percent predicted FEF25-75% (coefficient = -12.9, P = 0.01), and any granulomatous disease-like symptom [odds ratio (OR) = 3.1, 95% confidence interval (CI) = 1.45-6.73]. Mycobacteria (median = 658 CFU/m2 ) were positively associated with asthma symptoms (OR = 1.5, 95% CI = 0.97-2.43). Composite score (median = 11.5) of total bacteria from heat pumps was negatively associated with asthma (0.8, 0.71-1.00) and positively associated with FEV1 /FVC (coefficient = 0.44, P = 0.095). Endotoxin (median score = 12.0) was negatively associated with two or more granulomatous disease-like symptoms (OR = 0.8, 95% CI = 0.67-0.98) and asthma (0.8, 0.67-0.96). Fungi or (1→3)-ß-D-glucan in dust or heat pump traps was not associated with any health outcomes. Thermophilic actinomycetes and non-tuberculous mycobacteria may have played a role in the occupants' respiratory outcomes in this water-damaged building.

Detection of natural infection of infectious spleen and kidney necrosis virus in farmed tilapia by hydroxynapthol blue-loop-mediated isothermal amplification assay.

AIMS: Infectious spleen and kidney necrosis virus (ISKNV) has recently been recognized as a causative agent of serious systemic disease in tilapia. Our objective was to establish a new colorimetric loop-mediated isothermal amplification (LAMP) assay with pre-addition of hydroxynapthol blue (blue-LAMP) to investigate ISKNV transmission in tilapia. METHODS AND RESULTS: The blue-LAMP, targeting a major capsid protein gene of ISKNV, was conducted at 65°C for 45 min, allowing unaided visual detection of the pathogen based on colour change without cross-amplification of other known fish pathogens tested. Comparison of blue-LAMP and PCR assays revealed a higher detection level for blue-LAMP assay (41·33%) in a population of farmed tilapia infected with ISKNV. The investigation of ISKNV transmission pattern in farmed red tilapia using the blue-LAMP revealed a possible matroclinical form. The presence of ISKNV in the gonad samples was confirmed by in situ LAMP assay. Positive signals only appeared in ovarian follicles, and not in oocytes. Moreover, tissue tropism assay revealed that the brain was the main target organ in both farmed red tilapia (40%) and Nile tilapia (20%). CONCLUSIONS: The developed blue-LAMP assay has the potential to be used as a viable tool for screening covert and natural infections of ISKNV in tilapia. The evidence of vertical transmission of ISKNV infection in tilapia indicates the seriousness of this disease and will require a close attention and collaboration between tilapia hatcheries and disease experts in order to find a solution. SIGNIFICANCE AND IMPACT OF THE STUDY: The new blue-LAMP assay is a time-saving and economically viable detection tool, which allows unaided visual detection for ISKNV in tilapia, and it could be applicable for field applications. Evidence on the vertical transmission of ISKNV in farmed tilapia suggests a need for developing farm management practices to control the spread of virus in aquaculture industries.

Calibration of sharp cut impactors for indoor and outdoor particle sampling.

A low-flow rate, sharp cut point inertial impaction sampler was developed in 1986 that has been widely used in PM exposure studies in the United States and several other countries. Although sold commercially as the MS&T Area Sampler, this sampler is widely referred to as the Harvard Impactor, since the initial use was at the Harvard School of Public Health. Impactor nozzles for this sampler have been designed and characterized for flows of 4, 10, 20, and 23 L/min and cut points of 1, 2, 5, and 10 microns. An improved method for determining the actual collecting efficiency curve was developed and used for the recent impactor calibrations reported here. It consists of placing a multiplet reduction impactor inline just downstream of the vibrating orifice aerosol generator to remove the multiplets, thus allowing only the singlet particle s to penetrate through to the impactor being calibrated This paper documents the techniques and results of recent nozzle calibrations for this sampler and compares it with other size-selective inertial impactors. In general, the impactors were found to have sharp cutoff characteristics. Particle interstage losses for all of the impactors were very low, with the exception of the 10-micron cut size 20 L/min impactor, which had greater losses due to the higher flow rate. All of the 2.5-micron cut nozzle laboratory calibrations compare favorably to the U.S. Environmental Protection Agency (EPA) WINS-96 fine particle mass (PM2.5) impactor calibration data.

Air quality in an animal facility: particulates, ammonia, and volatile organic compounds.

Concentrations of ammonia, volatile organic compounds, particles, and mouse allergen were measured in an animal facility. Ammonia concentrations averaged less than 1 ppm, below any health-based standards. The concentrations of volatile organic compounds were in the 5-15 micrograms/m3 range. Among the volatile organic compounds found, only the terpenes a-pinene and a-terpinol (which may be derived from the pine shavings used as bedding) were consistently present in concentrations greater than outdoor air. The primary air contaminant present at concentrations high enough to be of known physiological significance was the mouse allergen, Mus ml. To determine which activities in an animal room generated the highest concentrations of airborne Mus ml, a monitor that counted particles continuously was used. The particle counts were correlated with allergen levels in the worker's breathing zone (r50.83,p,0.05). Thus, a particle counter can be used effectively in an animal facility to identify specific activities that generate high levels of both particles and allergen. Such activities included changing mice from soiled to clean cages, cleaning floors, and changing foam inserts in pressurized individually ventilated cages. To reduce exposure to allergen during cage changing, which is the major activity for an animal caretaker, a capture-type ventilated changing table was designed and tested. Use of such a table reduced exposure to allergen in the worker's breathing zone from 4.961.1 to 2.160.3 ng Mus ml/m3, a level comparable to background levels.

The effect of relative humidity on mouse allergen levels in an environmentally controlled mouse room.

To determine the effect of humidity on the levels of the mouse allergen Mus m 1, an experimental animal room was constructed to control environmental variables. The sex, strain, age, and number of mice was constant in the room, so that the average daily production of Mus m 1 would not vary greatly. Six different levels of relative humidity from 15% to 65% were maintained for a minimum of a week each. Daily collections of airborne particulates were eluted from filters and Mus m 1 content measured by immunological assay. Increasing relative humidity caused a decrease in Mus m 1 levels from a high of 3 ng/m3 at 15% humidity to a low of 0.5 ng/m3 at 65% humidity. Thus, reduction of airborne allergen levels can be achieved by careful attention to humidity control, especially during the winter heating season when humidity levels may be low. This experimental room can be used to measure the effect of other variables such as ventilation rate, caging, bedding, and work practices on the levels of mouse allergen in an animal facility.

Ventilation.

This chapter begins with an overview of the history of ventilation guidelines, which has led to the guidelines that are in effect today. Of particular interest is the most recent return in the past 5 years to ventilation rates that more closely reflect a mean or average of the range of guidelines that have existed over the past century. OSHA's and the EPA's recognition of the need to operate ventilation systems in buildings in an accountable manner is also of note. Of even more interest is the resurgence of the concept of minimum mixing and once-through ventilation air that has been pursued in parts of Northern Europe for the past 10 years, and in a school that is being designed with this concept in New Hampshire. In addition, the design concept of equipping office buildings with low pressure drop high efficiency particle filtration to remove fine particles from all of the air that is supplied to the occupants is being used increasingly in the U.S. This chapter also presents an overview of the various types of ventilation systems found in homes and commercial office buildings and the common indoor air quality problems that may be associated with them. It also offers an overview of common HVAC evaluation techniques that can be used to determine if a ventilation system is performing in a manner that makes sense for the use of the space and the needs of the occupants. Are the occupants receiving a reasonable supply of outdoor air? Is the air that they receive of reasonable quality? Are obvious pollutants being exhausted? Ventilation systems have become extremely complex and more difficult to run and maintain over the past 40 years. This trend will continue to drive the need for professionally maintained HVAC equipment that is serviced and run by individuals who are accountable for the quality of the air that the system delivers.

Retroperitoneal hematoma: an unusual complication of cold knife conization of the cervix.

A case of an unusual complication of cold knife conization and associated extensive retroperitoneal hematoma is presented. The contributing factors, prevention, and management of this complication are discussed.

Detection and quantitation of fetomaternal hemorrhage.

Failure to administer additional doses of Rh immune globulin (RhIG) to patients with excessive fetomaternal hemorrhage (FMH) is one of the causes of continued Rh isoimmunization. We compared the fetal cell ratio (FCR) with the other laboratory methods currently used at our hospital to quantitate FMH, ie, fetal cell preparation (FCP), RhIG cross-match with maternal serum, and indirect Coombs' testing 24 to 48 hours after administration of RhIG. The incidence of excessive FMH as detected by the various methods was 3.6% (cross-match), 9.6% (Coombs'), 18% (FCP), and 66% (FCR). The methods currently used do not accurately quantify FMH. The FCR is the most sensitive test but it has a high false-positive rate and thus does not appear to be clinically useful. We suggest that repeating the indirect Coombs' test may provide a practical alternative for determining the need for additional doses of RhIG.

Cytosol estrogen receptor content of female parietal peritoneum.

The ovarian and peritoneal mesothelium are believed to derive from the same embryonal coelomic epithelium. Neoplastic proliferation of these epithelia is sometimes grossly and histologically similar. Recently estrogen receptors have been identified in gynecologic neoplasms. With the use of standard techniques, the parietal peritoneum was assessed for estrogen receptor content in 17 women undergoing celiotomy for various reasons. All peritoneal specimens were negative for estrogen receptor content. This suggests a heterogeneous origin of the peritoneal surface epithelium and that peritoneum may not be müllerian in origin.

Neuroendocrine carcinoma of the uterine cervix complicated by pregnancy: case report and review of the literature.

A case report of neuroendocrine carcinoma of the cervix complicating pregnancy is presented. This represents the fourth reported case. Clinical staging was FIGO IB, and treatment consisted of radical surgery and aggressive chemotherapy. Electron microscopy confirmed the diagnosis of neuroendocrine carcinoma. The pregnancy-associated cases and a review of neuroendocrine cervical carcinoma are discussed.

Platelet fibronectin release induced by Walker 256 rat carcinoma tumor cells.

This study examined gel filtered rat platelet activation by Walker 256 rat carcinoma cells and characterized fibronectin release. Using aggregometry measurements, a biphasic platelet response was characterized and the timing of alpha granule release was determined. The localization and association of released platelet fibronectin with tumor cell-platelet aggregates was determined by immunofluorescent and immunocytochemical methods. The immunofluorescent studies showed that the platelet fibronectin was released and became associated with the peripheries of the tumor cells following alpha granule discharge. The ultrastructural immunocytochemical data revealed that this fibronectin was associated with a fibrin-like material, enmeshing the tumor cell-platelet aggregates. The significance of the release and localization of platelet fibronectin to tumor cell metastasis is discussed.

Cathepsin B activity in B16 melanoma cells: a possible marker for metastatic potential.

In solid s.c. tumors of a variant of the murine B16 melanoma with high metastatic potential (B16F10), there was a 2- to 7-fold elevation of lysosomal cathepsin B activity when compared to the B16F1 variant with low metastatic potential. The highest activities (based on either protein or DNA) of cathepsin B were found in tumors of less than 1 g. When B16F1 and B16F10 melanoma variants were grown in tissue culture, the metastatic differential in cathepsin B activity was lost as the cells were subcultured. However, this differential in cathepsin B activity could be restored by reestablishing the cultured cells as s.c. tumors. The activities of four other lysosomal enzymes (cathepsin D, beta-N-acetylglucosaminidase, beta-glucuronidase, and acid phosphatase) showed little evidence of a positive correlation with the metastatic potential of the B16 melanoma variants. Eighty to 90% of cathepsin B activity has been localized to a fraction containing viable tumor cells which was isolated by centrifugal elutriation. In contrast, only 50% of cathepsin D activity was in the viable tumor cell fraction, and from 30 to 70% of beta-N-acetylglucosaminidase, beta-glucuronidase, and acid phosphatase. Elevated levels of cathepsin B in the high metastatic B16F10 variant are consistent with the idea that cathepsin B may play a direct or a regulatory role in tumor metastasis.

A course in electronics and instrumentation.

In response to the continued development of electronic instrumentation in all disciplines of clinical laboratory science, we have designed a new format for teaching electronic instrumentation. This course provides basic concepts in electronics, including safety precautions, the use of electronic testing equipment, methods for determining precision and accuracy of instruments, interpretation of the operating manual, and preventive maintenance. Students apply concepts by actually maintaining and adjusting improperly functioning instruments, using systematic techniques for trouble-shooting and adjustment. Student examinations indicate that the objectives are being met.

Respiratory disease in Utah coal miners.

Two hundred forty-two Utah underground coal miners volunteered to participate in a respiratory disease study. They were an older group (mean, 56 years of age) and had spent a mean of 29 years in the coal-mining industry. The prevalence of chronic bronchitis was 57%, and that of coal worker's pneumoconiosis, 25%; only one worker had progressive massive fibrosis. Significant impairment of pulmonary function was found among those with a history of cigarette smoking. Chronic bronchitis or coal worker's penumoconiosis among nonsmokers did not impair pulmonary function. There was a significant association among the nonsmokers between increasing exposure to coal dust and coal worker's pneumoconiosis, but not for changes in pulmonary function. Coal mine dust had a significant influence in causing the symptom complex of chronic cough and sputum production, and coal worker's pneumoconiosis.

Common origin of pigment cells.

The fundamentally diverse vertebrate pigment cells, melanophores, xanthophores, and iridophores, contain pigmentary organelles known, respectively, as melanosomes, pterinosomes, and reflecting platelets. Their pigments are mealanins pteridines, and purines. Mosaic pigment cells containing more than one type of organelle have been observed and mosaic organelles containing more than one type of pigment have been discovered. It is proposed that the various pigment cells are derived from a stem cell that contains a primordial organelle of endoplasmic reticular origin. This primordial organelle can differentiate into any of the known pigmentary organelles.

The effects of lumicolchicine, colchicine and vinblastine on pigment migration in fish chromatophores.

The effects of lumicolchicine, colchicine, vinblastine and cold temperature on the pigment migration in melanophores and xanthophores of Fundulus heteroclitus and Oryzias latipes were examined by light and electron microscopy. Xanthophores of both species which contain numerous microfilaments and a poorly developed microtubule system were extremely sensitive to the alkaloids. Lumicolchicine and colchicine induced irreversible dispersion while vinblastine caused permanent aggregation of the pigments. Treatment in lumicolchicine or colchicine at 5 mM for 60 minutes did not disrupt microtubules of melanophores to an appreciable degree, however, melanosome aggregation was partially inhibited by these drug in Oryzias. When melanophores were kept in the cold in the presence of colchicine at 1 mM, almost all microtubules were disrupted and their repolymerization at room temperature was nearly completely inhibited by colchicine. These melanophores lacking in microtubules responded to epinephrine with slow aggregation. Vinblastine at 0.1 mM induced partial aggregation of melanosomes and disruption of microtubules but most melanophores were still able to respond with pigment migration. Vinblastine at 1 mM made all melanophores punctate and immobile. Large vinblastine-induced crystals were frequently seen in the dendritic processes. The results of the present investigation suggest that cytoplasmic microtubules in fish melanophores facilitate melanosome migration only in directional orientation and appear not be responsible for the motive force.