Visualization of episomal and integrated Epstein-Barr virus DNA by fiber fluorescence in situ hybridization

For many Epstein-Barr virus (EBV)-associated malignancies, it is still a matter of controversy whether infected cells harbor episomal or chromosomally integrated EBV genomes or both. It is well established that the expression of EBV genes per se carries oncogenic potential, but the discrimination between episomal and integrated forms is of great relevance because integration events can contribute to the oncogenic properties of EBV, whereas host cells that exclusively harbor viral episomes may not carry the risks mediated by chromosomal integration. This notion prompted us to establish a reliable technique that not only allows to unequivocally discriminate episomal from integrated EBV DNA, but also provides detailed insights into the genomic organization of the virus. Here, we show that dynamic molecular combing of host cell DNA combined with fluorescence in situ hybridization (FISH) using EBV-specific DNA probes facilitate unambiguous discrimination of episomal from integrated viral DNA. Furthermore, the detection of highly elongated internal repeat 1 (IR1) sequences provides evidence that this method permits detection of major genomic alterations within the EBV genome. Thus, fiber FISH may also provide valuable insights into the genomic organization of viral genomes other than EBV.     

Generalization of treatment effects following masturbatory training with erotic stimuli

Three married women were treated for situational orgasmic dysfunction in a laboratory-clinic through masturbatory training with erotic stimulation. Rate of orgasm was monitored via heart rate and verbal report. Treatment resulted in improved ability to attain orgasm by self-masturbation in the clinic and home. Husbands could not consistently stimulate their wives to orgasm until successive approximation programming with the couples was completed. This effect reportedly generalized from clinic to home and from masturbation to coitus. Follow-ups at two and six month intervals indicated maintenance of treatment effects.     

Acute rheumatic fever and rheumatic heart disease on the Navajo reservation, 1962-77.

The occurrence of acute rheumatic fever (ARF) and rheumatic heart disease (RHD) among Navajos was ascertained for the period from 1962 through 1977 by means of hospital discharge diagnoses and patients'' charts. The annualized attack rate for ARF was 12.4 per 100,000 population, with no clear evidence of an overall secular trend. The proportion of recurrences (19.6 percent) and clinical features were similar to those reported elsewhere, but no seasonal variation in attack rates was noted. Between 1962-71 and 1972-77, the age of RHD patients increased, suggesting few newly diagnosed cases and the aging of known patients. A streptococcal disease control program was instituted in many Navajo elementary schools before 1975. In the program, throat cultures were performed routinely for some asymptomatic children and for all symptomatic children. During the subsequent 3 years, ARF rates declined from 13.5 to 8.2 per 100,000 in areas covered by the program, while in the noncovered areas the rates showed little change-9.5 to 10.1 per 100,000.     

Immunization in the adult immunocompromised host

The number of patients with impaired immune response has been steadily increasing within the last years, not only with the onset of the AIDS epidemic, but also due to increasing numbers of subjects on immunosuppressive therapies. These patients are at an increased risk for infections, many of which are preventable by immunization. Inactivated vaccines are generally safe in subjects with underlying immunosuppression. However, immune response and protection may be hampered, depending on the extent of immunosuppression. In contrast, live vaccines such as yellow fever, measles, rubella, herpes zoster, and cholera may lead to severe reactions in immunocompromised patients and have been shown to deteriorate some immune-mediated diseases such as multiple sclerosis. Data on the efficacy of vaccines in biological therapies is scarce. Where necessary vaccines should be updated before immunosuppressive therapies are started. To improve the vaccination status several guidelines exist for immunosuppressed patients at risk such as those with rheumatic diseases, asplenia or solid organ and hematopoietic stem cell transplantation.