PCR amplification with modified primers, in combination with BcgI restriction fragment length polymorphism analysis, allowed us to differentiate a string of 7 Ts present in exon 7 of the ...21-hydroxylase gene of man from the 8 Ts present in exon 7 of its pseudogene. This approach can substitute for hybridization with oligonucleotides, or for sequence analysis, when an allele containing a multiple repeated nucleotide sequence is to be differentiated from one with a different number of the identical nucleotide.
Methods To address these issues, we used an eight-target tracking task, for which subjects used a joystick with their dominant hand in order to move a cursor positioned at the center of the screen to ...one of 8 targets following an elliptical trajectory (Fig1). Conclusions Our results support the hypothesis that time alone is sufficient to elicit savings in motor adaptation learning and that this sleep-independent process relies on the integrity of Lobule VI in the cerebellum.
The limited supple of appropriate tissues for study has been an impediment to investigations of varicella zoster virus (VZV) latency. Human dorsal root ganglia (DRG) harboring latent virus are not ...plentiful and are not amenable to manipulation for studying the events surrounding the establishment, maintenance, and cessation of latency. An alternative to studies in human DRG is the rat model of latency, which appears to provide a reliable method of investigating VZV latency. Other alternatives include studies in other human tissues involved in VZV pathogenesis. In order to improve our understanding of the establishment and cessation of latency, we performed comparative immunohistochemical analysis of chickenpox and zoster skin lesions. This analysis revealed that during primary infection and reactivation productive VZV infection occurs in a variety of cell types and that the major VZV DNA binding protein, ORF29p, is present in peripheral axons early during the course of chickenpox. VZV latency was studied in the rat model by in situ hybridization and compared with similar studies performed in human DRG containing latent virus, confirming that VZV DNA persists in the same sites in DRG of the two species.
The trans-Golgi network (TGN) is putatively the site where varicella-zoster virus is enveloped. gE is targeted to the TGN by selective retrieval from the plasmalemma in response to signaling ...sequences in its endodomain. gI lacks these sequences but forms a complex with gE. We now find that gI is targeted to the TGN and plasma membrane when expressed in Cos-7 cells; nevertheless, surface labeling revealed that gI is not retrieved from the plasma membrane. TGN targeting of gI depended on the T(338) of its endodomain and was lost when T(338) was deleted or mutated to A, S, or D. The endodomain of gI was sufficient, if it contained T(338), to target a fusion protein containing the ectodomain of the human interleukin-2 receptor to the TGN. A truncated protein consisting only of the gI ectodomain was secreted and taken up by nontransfected cells. This uptake of the secreted gI ectodomain was blocked by mannose 6-phosphate. Following cotransfection, both gI and gE were retrieved to the TGN from the plasma membrane in 26.7% of cells, neither gI nor gE was internalized in 18.3%, and gE was retrieved to the TGN while gI remained at the plasma membrane in 55%. We suggest that the T(338) of its endodomain is necessary to retain gI in the TGN; moreover, because gI and gE interact, the signaling sequences of each glycoprotein reinforce one another in ensuring that both glycoproteins are concentrated in the TGN yet remain on the cell surface.
To compare the prevalence of human papillomavirus (HPV) infections in women who are seropositive and seronegative for human immunodeficiency virus (HIV), and to determine if associations between HPV ...and cervical disease are altered in HIV-seropositive women.
In this cross-sectional study, 344 HIV-seropositive and 325 HIV-seronegative women underwent colposcopy and HPV DNA testing.
Human immunodeficiency virus-seropositive women were more likely than HIV-seronegative women to have HPV DNA of any type detected (60 versus 36%, P < .001). Infections with HPV type 16 (27 versus 17%, P < .05), type 18 (24 versus 9%, P < .05), and more than one type of HPV (51 versus 26%, P < .05) were also more common in HIV-positive women. Although both latent HPV infection and HPV infections associated with cervical intraepithelial neoplasia (CIN) were more prevalent in the HIV-seropositive group, the ratio between these two types of infections was altered markedly in the HIV-seropositive women. Human immunodeficiency virus-seropositive women who were HPV-infected were significantly more likely to have CIN than were HPV-infected HIV-seronegative women, an increase observed at all levels of immunosuppression. Analysis of specific HPV types associated with latent HPV infection and CIN indicated that HIV seropositivity only minimally alters the known associations between specific types of HPV and cervical disease.
Human papillomavirus infections are more common among HIV-seropositive women at all levels of immunosuppression. However, relationships between HIV and HPV are complex and cannot be explained completely by an increased susceptibility to new HPV infections in the immunosuppressed patient.
Background: When virologic and molecular diagnostic techniques are unavailable, the diagnosis of varicella zoster virus (VZV) infection depends on clinical criteria and histologic evaluation of skin ...biopsy specimens or Tzank preparations. These methods can misdiagnose chickenpox and zoster, particularly when the clinical manifestations are atypical.
Objective: To improve diagnosis in these settings, we developed an in situ hybridization technique for the detection of VZV utilizing a fluorescein-labeled oligonucleotide probe visualized with anti-fluorescein alkaline phosphatase-conjugated antibody.
Study design: We retrospectively examined 26 paraffin-embedded skin biopsy specimens with histologic features consistent with VZV or herpes simplex virus (HSV) infection and 11 control cases by in situ hybridization. In situ hybridization for VZV and HSV-1 was compared with polymerase chain reaction (PCR) for VZV and HSV-1 and clinical and histologic examination.
Results: Thirteen of the 26 study cases and two of the 11 control cases were positive for VZV by in situ hybridization. When compared with PCR, in situ hybridization was 92% sensitive and 88% specific. When compared with clinical diagnosis, in situ hybridization was 86% sensitive and 87% specific. All cases of chickenpox had VZV-positive inflammatory cells in the dermis but this finding was less frequent among the cases of zoster.
Conclusions: This in situ hybridization technique is a sensitive and specific method for the diagnosis of VZV in skin lesions that is applicable to most histopathology laboratory settings. In addition, in situ hybridization reveals individual infected cells and may provide insight into the pathogenesis of VZV skin infection.
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