Neuroendocrine,epigenetic, and intergenerational effects of general anesthetics

The progress of modern medicine would be impossible without the use of general anesthetics (GAs). Despite advancements in refining anesthesia approaches, the effects of GAs are not fully reversible upon GA withdrawal. Neurocognitive deficiencies attributed to GA exposure may persist in neonates or endure for weeks to years in the elderly. Human studies on the mechanisms of the long-term adverse effects of GAs are needed to improve the safety of general anesthesia but they are hampered not only by ethical limitations specific to human research, but also by a lack of specific biological markers that can be used in human studies to safely and objectively study such effects. The latter can primarily be attributed to an insufficient understanding of the full range of the biological effects induced by GAs and the molecular mechanisms mediating such effects even in rodents, which are far more extensively studied than any other species. Our most recent experimental findings in rodents suggest that GAs may adversely affect many more people than is currently anticipated. Specifically, we have shown that anesthesia with the commonly used GA sevoflurane induces in exposed animals not only neuroendocrine abnormalities (somatic effects), but also epigenetic reprogramming of germ cells (germ cell effects). The latter may pass the neurobehavioral effects of parental sevoflurane exposure to the offspring, who may be affected even at levels of anesthesia that are not harmful to the exposed parents. The large number of patients who require general anesthesia, the even larger number of their future unexposed offspring whose health may be affected, and a growing number of neurodevelopmental disorders of unknown etiology underscore the translational importance of investigating the intergenerational effects of GAs. In this mini review, we discuss emerging experimental findings on neuroendocrine, epigenetic, and intergenerational effects of GAs.     

Chlamydial infection of subcutaneous conjunctival transplants in guinea pigs

The development and testing of candidate vaccines for trachoma are constrained because only humans and nonhuman primates are susceptible to conjunctival infection with Chlamydia trachomatis. Guinea pig inclusion conjunctivitis (GPIC), an analogous disease of guinea pigs, provides a useful, less expensive model to study ocular chlamydial infections. GPIC is caused by a Chlamydia psittaci strain whose external constituents are very similar to those of C. trachomatis. To develop a better model for studying GPIC immunity, conjunctival pockets were established under the abdominal skin of guinea pigs by subcutaneous implantation. Up to six implants could be produced in each animal. The success rate of implantation was 79.0% (n = 148). These pockets were then infected with GPIC. The organism was recovered from the autografts indicating local replication, and tests for serum antibody by microimmunofluorescence showed production of GPIC-specific antibody of IgG and IgM classes after infection. There was minimal antibody response after moderate inoculating doses to the implants, and the titers increased more slowly than after eye infection with GPIC; with higher concentration of the inoculum, however, the antibody response increased to the same levels as with the ocular challenge but more slowly. Inoculation of pockets with GPIC also produced acute inflammatory changes in infected autografts (n = 101). Histologic examination of infected grafts showed chlamydial inclusions in epithelial cells and significant infiltration with lymphocytes and polymorphonuclear cells. Subcutaneous autografts may provide a useful model for chronologic studies of chlamydial infection. The delayed immunologic response, however, suggests that these pockets of implanted epithelium do not have full access to the immune system.     

Respiratory symptoms and ventilatory capacity in swine confinement workers.

A group of 59 workers (41 men and 18 women) employed in swine confinement areas was studied to assess the presence of acute and chronic respiratory symptoms and the prevalence of abnormalities in ventilatory function. A control group of 46 (31 men and 15 women) unexposed workers was studied for the prevalence of chronic respiratory symptoms. For both male and female swine confinement workers complaints of chronic cough, dyspnoea, and chest tightness were significantly more frequent than among control workers. Male workers also complained more of chronic phlegm. Male swine confinement workers who were smokers had significantly higher prevalences of chronic cough, chronic phlegm, and chronic bronchitis than male non-smoking swine confinement workers. The frequency of acute symptoms associated with the workshift was high among the swine confinement workers with more than half of the workers complaining of cough and dyspnoea associated with work. Significant acute across shift reductions in lung function occurred in swine confinement workers, being largest for FEF25. All Monday preshift ventilatory capacity measurements in male confinement workers were significantly lower than predicted values; FVC and FEV1 were found to be lower than predicted values for women. The data indicate that exposure in swine confinement buildings is associated with the development of acute and chronic respiratory symptoms and impairment of lung function. Smoking appears to aggravate these changes.     

Evaluation of Syva's enzyme immunoassay for the detection of Chlamydia trachomatis in urogenital specimens.

A newly developed microwell enzyme immunosorbent assay (EIA) system by Syva Company (Palo Alto, CA) can detect Chlamydia trachomatis in < 3 hr. It uses a polyclonal antibody to chlamydial lipopolysaccharide and end points are determined with a spectrophotometer. Three clinical trial sites (University of California Medical Center, San Francisco, CA; University of Washington, Seattle, WA; and Louisiana State University Medical Center, New Orleans, LA), compared this EIA with tissue culture (TC) for identifying Chlamydia in urogenital specimens. Overall prevalence by TC was 10.4% (136 of 1306). When tests were compared with TC (using vials or microtiter plates and a fluorescent antibody stain), we found an EIA sensitivity of 93.4% (127 of 136) and a specificity of 98.1% (1148 of 1170). This EIA has a performance profile that is, at the very least, comparable with other nonculture methods for diagnosing genital tract infections with C. trachomatis.     

Immunological and respiratory reactions in workers exposed to organic dusts

The relationship of skin reactivity and serum immunoglobulin E (IgE) levels to the prevalence of chronic respiratory symptoms and to ventilatory capacity is examined in workers exposed to different organic aerosols. The results from group of control workers similarly tested are also presented. Workers exposed to occupational allergens had positive skin tests more frequently than did controls, except for soy bean workers. Workers with positive skin tests to occupational allergens had a higher prevalence of almost all symptoms than those with negative skin tests although the differences did not always reach statistical significance. Workers with positive skin reactions in general had significantly higher serum IgE levels than did workers with negative skin reactions. There were across-shift reductions of ventilatory capacity in all groups of exposed workers, varying for forced vital capacity from 1.7% to 13.3%, for forced expiratory volume from 0.4%–21.9%, for maximum flow rates at 50% from 1.5% to 16.1% and for maximum flow rates at the last 25% of control vital capacity from 0% to 24.9%. There was, however, no correlation between acute and chronic lung function changes and skin reactivity or IgE values. Our data suggest that although exposure to organic aerosols may be associated with frequent immunologic reactions, these findings do not predict objective respiratory impairment.The research was supported in part by grant no. JBP 733 from the National Institutes of Health, Bethesda, Md., USA, and by grant no. RO1 OHO-2593-04 from the National Institutes of Occupational Safety and Health, Centers for Disease Control, Atlanta, Ga., USA     

Acute glomerulonephritis secondary to lobar pneumonia

We report 4 cases of children who developed lobar pneumonia and subsequently acute glomerulonephritis, all within a period of six weeks. In one case streptococcus pneumonia type 5 was isolated, a strain not previously described in the literature as being nephritogenic.     

Cocaine decreases self-control in rats: a preliminary report

Cocaine abuse is often associated with behavior that takes into account short-term, but not long-term consequences. However, there has been no empirical research concerning the effects of cocaine on self-control (choice of a larger, more delayed reinforcer over a smaller, less delayed reinforcer). In the present research, when food-deprived rats repeatedly chose between a larger, more delayed food reinforcer and a smaller, less delayed food reinforcer, chronic intraperitoneal injections of 15 mg/kg cocaine (but not 10 mg/kg fluoxetine) decreased the rats' choices of the larger, more delayed reinforcer. Cocaine can decrease rats' self-control.     

Optimization of non-isotopic in situ hybridization on formalin-fixed, paraffin-embedded material using digoxigenin-labelled probes and transgenic tissues.

The sensitivity of non-isotopic in situ hybridization (NISH), particularly on formalin-fixed, paraffin-embedded (FFPE) clinical tissues, has been the subject of controversy. Generally, NISH has been regarded as being less sensitive than radiolabelled procedures, although some reports have contradicted this. Accordingly, tissues from mice which were transgenic for variable amounts of the human alpha-1-antitrypsin gene were used to optimize the NISH procedure and to estimate the sensitivity. This approach showed that prolonged incubation of slides in final substrate resulted in high sensitivity--about 13 kb of target DNA. However, this prolonged incubation crucially depended on achieving minimal non-specific background staining. Many factors affected the degree of background staining, but five were particularly important. First, the method of mounting cut sections onto slides. Second, the length of the probe (ideally less than 400 bp). Third, the procedure for proteolytic digestion. Fourth, the denaturation technique, and fifth, the quality of the dextran sulphate used in the hybridization mix. The optimized protocol showed variable patterns of mRNA distribution in the transgenic mouse livers, while DNA distribution appeared uniform.