Detection of Borrelia burgdorferi in human blood and urine using the polymerase chain reaction.

We investigated the use of the polymerase chain reaction (PCR) to detect Borrelia burgdorferi strain B-31 in human blood and urine experimentally inoculated with 5 and 1 borreliae/cm3, respectively, and to biotinylate a DNA probe specific for B. burgdorferi in the dot blot and Southern blot assays. When the blood and urine samples were subjected to PCR, a 370-bp amplified product was consistently visible on agarose gel electrophoresis after 30 and 45 cycles, respectively. The total human genomic DNA extracted from a 1-cm3 sample of inoculated blood was approximately 6.25 micrograms, and the total amount of B. burgdorferi DNA was estimated to be 0.01 pg/6.25 micrograms of the human DNA. For PCR, 2.5 micrograms of human DNA which contained the equivalent of 0.004 pg of borrelia DNA (approximately two borreliae) were used for enzymatic amplification. When 1/20 or 1/10 of the PCR-amplified products were used either for dot blot or Southern blot hybridization, the accessible copies of amplified B. burgdorferi DNA were sufficient for detectable hybridization to occur. PCR amplification of B. burgdorferi DNA in clinical specimens followed by dot blot hybridization may be a valuable adjunct or alternative to current but inadequate laboratory methods for the diagnosis of Lyme disease.     

Detection of parvovirus B19 DNA, antigen, and particles in the human fetus

Human parvovirus B19 commonly infects children, causing erythema infectiosum (fifth disease). However, there is a significant adult population which has not been exposed to the virus and, consequently, does not have protective antibody. Recent reports have associated B19 infection during pregnancy with fetal death, although normal outcome of pregnancy is more common. To characterise further the role of B19 infection in fetal deaths, a series of laboratory investigations has been undertaken on tissues obtained at autopsy. These have demonstrated the presence of virion-sized DNA by Southern blotting, viral antigen by radioimmunoassay, and viral particles by electron microscopy, all from tissues of hydrops fetalis. These data confirm that the human parvovirus B19 can cross the placenta and replicate in fetal tissues.     

Myositis associated with a newly described microsporidian, Trachipleistophora hominis, in a patient with AIDS.

Microsporidia are zoonotic protozoa which were rare human pathogens prior to 1985, when Enterocytozoon bieneusi was described in human immunodeficiency virus-infected patients with chronic diarrhea. Another species, Encephalitozoon (Septata) intestinalis, is associated with diarrhea and chronic sinusitis, and approximately 25 cases have been reported in the literature. However, other microsporidial infections in human immunodeficiency virus-infected patients remain extremely rare. We report the first case of a Pleistophora sp.-like microsporidian infection presenting as a progressive severe myosotis associated with fever and weight loss. The organism was demonstrated by light microscopy and electron microscopy in corneal scrapings, skeletal muscle, and nasal discharge. Electron microscopy showed an electron-dense surface coat with "sunflare"-like projections surrounding all stages of development of meronts (two to four nuclei, dividing by binary fission), sporonts, and sporoblasts. Division of sporonts, in which sporonts separate from the thick outer coat, creating a sporophorous vesicle, is by binary fission, differentiating this organism from Pleistophora sp. The spore measures 4.0 by 2.5 microns and has a rugose exospore. A new genus and species, Trachipleistophora hominis, has been established for this parasite. The patient was treated with albendazole, sulfadiazine, and pyrimethamine, and the clinical symptoms resolved.     

Ensheathment of the olfactory nerves in the adult rat

The ensheathment of the olfactory nerve fibres is achieved by cooperation of two cell types. The olfactory ensheathing cells have a rounded outer surface enclosed in a continuous single basal lamina, and enclose an inner compartment from which overlapping processes of the same and adjacent cells enwrap interweaving territories of tightly apposed aligned axons. The olfactory nerve fibroblasts are highly flattened, dense cells generating multiple layers of very thin processes encircling individual or groups of olfactory ensheathing cells. This paper illustrates the unique ultrastructural features of this ensheathment.     

Relative right versus left frontal EEG in neonates

Although infants have been noted to have greater relative right or left frontal EEG as early as the neonatal period, other ways in which these newborns differ have not been reported. In this study, 48 newborns were divided on the basis of greater relative right versus greater relative left frontal EEG to determine whether these groups differed in other ways at the neonatal period including behavior, physiology, and biochemistry. We also were interested in whether these EEG patterns were related to any prenatal maternal variables including mood states (depression, anxiety, anger) and biochemistry as well as fetal activity. The greater relative right frontal EEG newborns had mothers with lower prenatal and postnatal serotonin and higher postnatal cortisol levels. The mothers of the greater relative right frontal EEG newborns also had greater relative right frontal EEG activation and lower vagal tone. The greater relative right frontal EEG newborns themselves had elevated cortisol levels, showed a greater number of state changes during sleep/wake behavior observations, and performed less optimally on the Brazelton Neonatal Behavior Assessment (T. B. Brazelton, 1973) including the habituation, motor, range of state, excitability, and depressive symptoms scales. These data suggest that greater relative right frontal EEG newborns may be at greater risk for developmental problems than those with greater relative left frontal EEG activation. In addition, a discriminant function analysis correctly classified 67% of the newborns' EEGs by prenatal maternal variables, suggesting that these might be used to target pregnant women for prenatal intervention.     

The diagnosis of recent herpes simplex virus type 2 genital infections by the simplex-2 test.

The prevalence of complement-fixing (CF) antibody against the AG-4 early antigen of herpes simplex virus (HSV) type 2 (HSV-2) was determined in patients with culture confirmed HSV-2 genital herpes and control groups using a commercial HSV-2 early antigen (Simplex-2; Gene Link Australia Ltd). Eighty seven per cent of 39 sera collected between 14 and 28 days after confirmed primary and recurrent HSV-2 infection were positive. In acute sera collected between 2-10 days after onset the Simplex-2 test was negative in all 90 patients with presumed primary infection but positive in 53% of 230 sera from recurrent infection. A specificity of 90-94.5% was obtained by testing 36 patients with recent proven HSV-1 infection and 331 control group patients. The Simplex-2 test may be useful in some cases of culture-negative, clinically suspected genital HSV-2 lesions only when sera are collected between 14-28 days after primary and recurrent infection. Its lack of specificity makes it unsuitable for the routine diagnosis of recent HSV-2 infection in the general population.     

Role of Peripheral Adrenergic Responsiveness in the Development of DOCA/NaCl Hypertension in Rats

Alterations in - and -adrenergic responsiveness were investigated prior to and during the development of hypertension in rats treated with desoxycorticosterone acetate and NaCl (DOCA/ NaCl). The DOCA/NaCl rats became noticeably hypertensive (> 150 mm Hg) six weeks after the initiation of treatment. Prior to the development of hypertension, a reduced in vivo and in vitro - and an enhanced -adrenergic responsiveness of the DOCA/NaCl group resulted. At 2 and 12 weeks of the study, the dipsogenic response to isoproterenol was significantly attenuated in the DOCA/NaCl rats, whereas no difference in the dipsogenic response to 24 hour water deprivation was observed between control and DOCA/NaCl rats. Isoproterenol-induced relaxation of aortic smooth muscle from the DOCA/NaCl treated rats was significantly reduced at 4 weeks and further attenuated at 12 weeks of the study. However, aortic smooth muscle sensitivity to norepinephrine stimulation was significantly increased at 4 and 12 weeks of the study. These results suggest that alterations in both in vivo and in vitro - and -adrenergic responsiveness occur prior to establishment of hypertension of the DOCA/NaCl rats and that these alterations may have a role in the early stages of the development of this form of hypertension.     

Chemonucleolysis. A ten-year retrospective study

An 11.8-year median follow-up evaluation of 42 "ideal" patients who had chemonucleolysis was obtained by examination, questionnaire, and roentgenograms. The excellent and good rating of this group was 81%, as compared to the total of 135 patients previously evaluated at 42 months and showing 85.2%. No complications were noted. Disc space rewidening, after initial narrowing, was observed in eight patients and 26% of all discs injected. While all who showed rewidening had excellent results, widening was not necessary to achieve an excellent rating. Marked narrowing and sclerosis of intervertebral margins were seen in many excellent and good clinical results. Chemonucleolysis represents a viable option as definitive treatment for a herniated nucleus pulposus in carefully selected patients.