Effect of primer selection on estimates of GB virus C (GBV-C) prevalence and response to antiretroviral therapy for optimal testing for GBV-C viremia.

GB virus C (GBV-C; also called hepatitis G virus) is a common cause of infection associated with prolonged survival among HIV-infected individuals. The prevalences of GBV-C viremia vary widely in different studies, and there has been poor agreement among different laboratories performing GBV-C RNA detection in quality control studies. To determine the optimal method of measuring GBV-C RNA in clinical samples, samples obtained from 939 HIV-infected subjects were studied using reverse transcription (RT)-PCR methods amplifying four separate regions of the GBV-C genome. Primers amplifying the E2 coding region were 100% specific; however, their sensitivity was only 76.6%. In contrast, primers amplifying three additional conserved regions of the GBV-C genome (the 5' nontranslated region and the nonstructural protein-coding regions 3 and 5A) were more sensitive but produced higher rates of false-positive results. Using low-specificity primer sets influenced the significance of association between GBV-C viremia and response to antiretroviral therapy. Using a quantitative GBV-C RNA method, the GBV-C RNA concentration did not correlate with baseline or set point HIV RNA levels; however, a correlation between negative, low, and high GBV-C RNA levels and increasing reduction in HIV RNA following antiretroviral therapy was observed. Subjects with both GBV-C E2 antibody and viremia had significantly lower GBV-C RNA levels than did viremic subjects without E2 antibody. These studies demonstrate that accurate detection of GBV-C RNA by nested RT-PCR requires the use of primers representing multiple genome regions. Analyses based on testing with single primers do not lead to reliable conclusions about the association between GBV-C infection and clinical outcomes.

Effect of GB virus C on response to antiretroviral therapy in HIV-infected Brazilians.

OBJECTIVES: GB virus C (GBV-C) infection is associated with delayed mortality in HIV-infected people in most, but not all, studies. Previous investigations of the effect of GBV-C viraemia on response to antiretroviral therapy (ART) were inconclusive. To determine the effect of GBV-C on ART, we retrospectively analysed plasma samples taken from patients in a prospective randomized clinical trial of ART in HIV-positive Brazilians. METHODS: GBV-C viraemia was characterized by testing stored serum samples from 175 participants by reverse transcriptase-polymerase chain reaction (RT-PCR). Subjects were randomized to receive indinavir (n=59), zidovudine and lamivudine (n=58), or zidovudine, lamivudine and indinavir (n=58). The effect of GBV-C viraemia on the average change in HIV viral load and CD4 count following initiation of therapy was evaluated in a multiple regression analysis. RESULTS: The prevalence of GBV-C viraemia was similar to that observed in previous studies (24%). HIV viral load decreased following ART to a significantly greater extent in patients with GBV-C viraemia (by 0.48 log(10) HIV-1 RNA copies/mL, P=0.009, adjusting for age, ART group, and baseline CD4 count). Although there was no significant difference in change in CD4 count between individuals with and without GBV-C viraemia overall, CD4 counts were higher following 48 weeks of therapy in GBV-C viraemic individuals receiving the least potent ART regimen (zidovudine and lamivudine) compared with those without GBV-C infection. CONCLUSIONS: GBV-C viraemia is associated with an enhanced reduction of HIV viral load in response to ART. In this study of treatment-naive individuals during 48 weeks of follow up, patients with GBV-C viraemia had reductions in HIV viral load that were approximately 0.5 log copies/mL greater than those found in patients without GBV-C viraemia. This is similar to reductions observed with nucleoside reverse transcriptase inhibitors.

Long-term potentiation in the rat dentate gyrus is associated with enhanced Arc/Arg3.1 protein expression in spines, dendrites and glia.

Electron microscopic immunocytochemical methods were used to determine the localization, subcellular distribution and expression of activity-regulated cytoskeletal protein (Arc/Arg3.1) in dentate gyrus after unilateral induction of long-term potentiation (LTP) in the perforant pathway of anaesthetized rats. At 2 h post-induction, immunoreaction product was visible in the dentate gyrus in both the granule cell and molecular layers. Arc expression was higher in the potentiated than the unstimulated contralateral hemisphere. Single-section electron microscopy analysis in unstimulated tissue and in tissue prepared 2 and 4 h after LTP induction showed Arc immunoreactivity (Arc-IR) in dendrites, dendritic spines and glia. Arc-IR was associated with synaptic and non-synaptic plasma membrane apposed to axon terminals and with cytoplasmic organelles, including the cytoskeleton. Arc-IR was also present in neuronal perikarya and there was occasional labelling of nuclei and axons. At 2 h post-LTP induction, there were significant increases in Arc-IR within the granule cell and molecular layers of the dentate gyrus and particularly within the middle molecular layer relative to the inner and outer molecular layers. This increase in Arc expression 2 h after LTP induction was blocked by the N-methyl-D-aspartate receptor antagonist (RS)-3-2-carboxypiperazin-4-yl-propyl-1-phosphonic acid. In animals killed 4 h after LTP induction, Arc expression had declined and differences between the potentiated and unpotentiated hemispheres were no longer significant. Our data provide ultrastructural evidence for a transient LTP-associated increase in the expression of Arc protein in the middle molecular layer of the dentate gyrus, with preferential targeting to dendrites, dendritic spines and glia.

Detection and typing of human papillomavirus in cutaneous warts of patients infected with human immunodeficiency virus type 1.

BACKGROUND: Cutaneous warts are caused by human papillomavirus (HPV). To date, more than 120 different types of HPV are known, of which 80 have been completely characterized. Prevalence studies on types of HPV present in cutaneous warts have been carried out in immunocompetent individuals and immunosuppressed organ allograft recipients, but not in human immunodeficiency virus (HIV)-positive patients. OBJECTIVES: To determine the HPV types present in cutaneous warts of HIV-infected patients. METHODS: Twenty-five biopsies of cutaneous warts from HIV-infected patients and 14 samples from control non-HIV-infected patients were studied. HPV detection was performed by polymerase chain reaction using two sets of primers: MY09/MY11 and RK91. The type of HPV was determined by restriction fragment length polymorphism analysis and direct sequencing of the amplified products. RESULTS: HPV DNA was detected in 64% of cutaneous warts from HIV-infected patients and in 79% of samples from the control group. The HPV types identified in HIV-infected patients were: HPV 2 (38%), 57 (31%), 27 (12%), 6 (12%) and 7 (6%). HPV 2/27/57 predominated in both groups, being present in 81% of lesions from HIV-infected patients and 82% of samples from non-HIV-infected patients. HPV 6, a genital HPV type rarely found in cutaneous lesions, was detected in two warts from HIV-infected patients and in one lesion of the immunocompetent group. HPV 7, characteristically associated with butcher's warts, and recently detected in oral and perioral lesions of HIV-infected patients, was found for the first time in a non-facial lesion of an HIV-infected patient. CONCLUSIONS: This is the first study evaluating the prevalence of HPV types in cutaneous warts of HIV-infected patients and immunocompetent individuals in Brazil.

Late hemorrhagic disease of newborn.

The clinical features of 14 infants diagnosed with late hemorrhagic disease of newborn (LHDN), of which 10 did not receive vitamin K prophylaxis, are presented. All infants were exclusively breast-fed and 12 did not have any underlying illness to explain the abnormal coagulation profile. The common presenting symptoms were seizures (71%), vomiting (57%), poor feeding (50%) and altered sensorium (36%). Physical examination shared pallor in all infants and a bulging anterior fontanel in 64%. Intracranial bleed was the predominant manifestation (93%), with CT scan showing intracranial bleed in 78%. Eight infants (57%) succumbed to their illness, while 36%had neurological sequelae. Since LHDN leads to significant morbidity and mortality, it should be prevented by providing vitamin K prophylaxis to all newborns.

Persistent alterations in dendrites, spines, and dynorphinergic synapses in the nucleus accumbens shell of rats with neuroleptic-induced dyskinesias.

Chronic treatment of humans or experimental animals with classical neuroleptic drugs can lead to abnormal, tardive movements that persist long after the drugs are withdrawn. A role in these neuroleptic-induced dyskinesias may be played by a structural change in the shell of the nucleus accumbens where the opioid peptide dynorphin is upregulated in treated rats that show vacuous chewing movements (VCMs). The shell of the nucleus accumbens normally contains a dense plexus of dynorphinergic fibers especially in its caudomedial part. After 27 weeks of haloperidol administration and 18 weeks of withdrawal, the immunoreactive labeling of this plexus is intensified when compared with that after vehicle treatment. In addition, medium spiny neurons here show a significant increase in spine density, dendritic branching, and numbers of terminal segments. In the VCM-positive animals, the dendritic surface area is reduced, and dynorphin-positive terminals contact more spines and form more asymmetrical specializations than do those in animals without the syndrome (VCM-negative and vehicle-treated groups). Persistent, neuroleptic-induced oral dyskinesias could therefore be caused by incontrovertible alterations, involving terminal remodeling or sprouting, to the synaptic connectivity of the accumbal shell.

Chronic haloperidol treatment impairs glutamate transport in the rat striatum.

High-affinity, Na(+)-dependent transport of glutamate into neurons and glial cells maintains the extracellular concentration of this neurotransmitter at a sub-toxic level. Chronic blockade of dopamine D(2) receptors with haloperidol elevates extracellular glutamate levels in the striatum. The present study examines the effect of long-term haloperidol treatment on glutamate transporter activity using an assay based on measuring the uptake of D-[3H]aspartate in striatal synaptosomes prepared from male Wistar rats. The maximal rate of glutamate transport in the striatum is reduced by 63% following 27 weeks of haloperidol treatment. This impairment of glutamate transport may be important in chronic neuroleptic drug action.

NMDA receptor blockade attenuates the haloperidol induction of Fos protein in the dorsal but not the ventral striatum.

Neuroleptic blockade of dopamine receptors is known to produce an increase in the expression of Fos. This increase may be related to elevations in glutamate transmission which in turn activates N-methyl-D-aspartate (NMDA) receptors. In the present study, we examine the role of these receptors in the haloperidol-induced augmentation of Fos in the caudate-putamen and nucleus accumbens of Wistar rats. Animals were divided into four groups for each experiment and each was injected either with saline; a noncompetitive NMDA antagonist, dizocilpine maleate (MK801, 5 mg/kg); haloperidol (0.5 mg/kg); or MK801 followed by an injection of haloperidol. Fos-immunoreactive cells appear in large numbers in all parts of the striatum 3 h after the administration of haloperidol. Pretreatment with MK801 attenuates the haloperidol-induced increase in Fos in the caudate-putamen. However, antagonism of the NMDA receptor does not significantly reduce the density of Fos-immunoreactive cells in any territory of nucleus accumbens, i.e., shell, core, or rostral pole. These data suggest that haloperidol acts in an NMDA-dependent manner in the caudate-putamen, but independently in parts of nucleus accumbens traditionally considered to be targets of antipsychotic drugs.

Cytomegalovirus transmission in child care homes.

BACKGROUND: Children attending child care centers have high rates of cytomegalovirus (CMV) excretion. Women exposed to such children have an increased risk of acquiring CMV infection, and primary infection places the offspring of such women at risk of congenital CMV infection. We studied family child care homes to determine if this child care alternative might represent a safe haven from CMV. METHODS: One hundred thirty-two women providing care in their homes were studied using a latex agglutination method to determine the rate of CMV seropositivity at baseline. Women who were seronegative for CMV were then sampled prospectively at 6-month intervals between March 1991 and August 1994 to determine the annual rate of CMV acquisition. A point prevalence of CMV excretion in family homes was determined by sampling 106 children from 25 randomly selected homes. Cytomegalovirus isolates were compared by molecular analysis using polymerase chain reaction-based methods to identify transmission. RESULTS: At baseline, 57.6% of the 132 providers were seropositive for CMV. Seropositive providers were more likely to be caring for toddlers (aged 1-2 years) (67% vs 46%; P=.02) and had worked in child care somewhat longer (median of 28.5 vs 21.5 months; P=.11). Using stepwise logistic regression, the strongest predictors of seropositivity at baseline were caring for children aged 1 to 2 years (odds ratio [OR] =2.37; P=.02) and number of months as a child care provider (OR= 1.17 for an increase of 24 months as provider; P=.08). Six or more years as a provider was highly associated with seropositivity (OR=3.27; P=.02). During follow-up, 5 of 51 seronegative providers seroconverted, yielding an annual infection rate of 6.8%. The point prevalence survey of children from the 25 homes (14 had seropositive providers) identified 8 CMV-excreting children. Three children in 1 home had indistinguishable isolates by polymerase chain reaction mapping. The provider seroconverted and excreted an isolate with a molecular profile indistinguishable from that of the children. CONCLUSIONS: The prevalence of CMV excretion is low among children attending child care homes (8% vs 15% in prior studies of child care centers; P=.07), and only 1 (20%) in 5 of the homes had CMV-excreting children. However, the overall CMV seroconversion rate of home child care providers was comparable to the rate observed among providers in child care centers. Families who use family home child care as an alternative to large child care centers are exposed to a low and unpredictable risk of CMV infection.

Epidemiology of congenital cytomegalovirus infection: maternal risk factors and molecular analysis of cytomegalovirus strains.

To determine factors that influence the occurrence of congenital cytomegalovirus (CMV) infection, the authors surveyed prospectively 8,254 infants born in eastern Iowa between October 1989 and June 1994. The authors conducted a case-control study to identify maternal risk factors, matching each CMV-infected infant with three uninfected infants according to hospital and date of birth. CMV strains were compared by using the polymerase chain reaction (PCR) to identify common sources of infection. Of the 7,229 infants cultured successfully for CMV, 35 (0.48%) were congenitally infected. Mothers of CMV-infected infants were more likely to be single (odds ratio (OR) = 3.05, p = 0.016), to work in sales (OR = 4.93, p = 0.008), or to be students (OR = 5.01, p = 0.017). Conversely, women who worked in health-care professions were less likely to have a congenitally infected infant (OR = 0.14, p = 0.049). PCR analysis indicated 27 distinct strains of CMV, but two groups of infants (two infants per group) excreted strains with indistinguishable molecular patterns. One of these pairs of infants had older siblings who attended the same child-care center during their mothers' pregnancies. The authors concluded that demographic and occupational factors influenced the risk of giving birth to an infant with congenital CMV infection. Many distinct CMV strains were identified, suggesting that major point source outbreaks had not occurred. Nonetheless, point source acquisition of CMV from child-care environments did account for some cases of congenital CMV infection in eastern Iowa.

[Care of women under chemotherapy treatment. Comprehensive analysis of care]. / Enfermagem à mulher em tratamento quimioterápico--uma análise compreensiva do assistir.

This study was developed upon my reflections as a nurse working at the Antineoplastic Chemotherapy Center at Luiza Gomes de Lemos Hospital of the National Cancer Institute (Inca), and as a Master course student at Anna Nery Nursing School. Developing my Master thesis on Martin Heidegger's philosophical thinking it has enabled me to reflect about the nursing care, considering this philosopher's thinking. In this study, I have registered some situations that occurred on my working days, in the scenery mentioned above, and reflected about them, seeking to analyze their proximity and distance from Heidegger's philosophical thinking. This study was also a reflection about questions emerging from the research process and care practice which leaded me to another understanding of nursing care to women who are submitted to chemotherapy treatment. Then, I was able to see nursing care in another way, that one of comprehension as a possibility of professional action.

[Phenomenology as a methodologic reference: sharing the experience of women who search for the prevention of cervical cancer]. / A fenomenologia como abordagem metodológica: compartilhando a experiência de mulheres que buscam a prevenção do câncer cérvico-uterino.

The article has the purpose to bring out the experience of having phenomenology as a methodological reference and Martin Heidegger's philosophical thinking expressed in the book entitled ¿Being and Time, used by nursing in order to understanding women who search for the prevention of cervical cancer as well as to analyse teh programs offered to women.

[An approach to the phenomenological interview]. / Um caminhar na aproximação da entrevista fenomenológica.

The purpose of this study is to describe my experiences about the phenomenological interview. It was developed in four moments, in order to adequate physical environment aspects, strategies of approaching the interviewers, the orientation question and the techniques to obtain the statements. The focus on researcher-subject/researched-subject has indicated adjustments in the utilization of the techniques and reoriented my person and professional development.

Cytomegalovirus infection in newborns and their family members: polymerase chain reaction analysis of isolates.

Polymerase chain reaction (PCR) analysis with primers for the pp65, a-sequence, glycoprotein B, and major immediate early genes of human cytomegalovirus (CMV) was used to study five congenitally-infected infants and their CMV-infected family members. Family members excreting CMV included three mothers and two siblings. The PCR results indicated that the CMV strain excreted by each infant was indistinguishable from that excreted by the corresponding family member. By contrast, the molecular profiles of the CMV strains were distinct between families, indicating that the PCR algorithm described in this study is a useful method for analyzing CMV strains.

[Experience of women in labor. Nurses' observations]. / Vivência de parturientes: observação de enfermagem.

This study emerged from my own experience in Obstetric Nursing and my living in Obstetrics. I searched for comprehending the real meaning of delivery labor experiences as a situated phenomenon. It is a qualitative research with phenomenological approach. The lying-in women were the witnesses through the question: 'What was your daily experience in the pre-delivery room like?' Through analysis and hermeneutics of speeches, I have used the knowledge of Matin Heidegger's philosophical-theoretical referential. Thus, I managed to clarify that the parturient is fearful when left by herself in her experience, showing surprise for the labor of delivery is singular, noting down the pain as a possibility of the existential limit, being open to help and noticing the impersonality of care provided. With this watchful sense, I approached the comprehension of a lying-in woman as a being-in-a-world-with-health-professionals. This discovery indicated the possibility of new ways of caring in Obstetrics.

Cytomegalovirus reinfection in young children.

OBJECTIVE: To determine the frequency of reinfection with new cytomegalovirus (CMV) strains in children in group child-care environments. METHODS: Ninety-two CMV strains isolated serially from children attending child care centers were analyzed. Strains were obtained from 1986 to 1994, from 37 children attending one of six centers in the area of Cedar Rapids and Iowa City, Iowa. The CMV isolates were analyzed by a polymerase chain reaction-based algorithm using primers for the a-sequence, glycoprotein B, and major immediate early (MIE) genes of human CMV. The a-sequence polymerase chain reaction products were compared on the basis of size, and products derived from glycoprotein B and MIE genes were compared according to restriction fragment length polymorphisms. RESULTS: Children were between 8 months and 5 years 7 months of age at the time of CMV isolation. The number of isolates ranged from 2 to 6 per child, and the intervals between the first and last CMV isolation ranged from 11 weeks to more than 3 years. At least 7 (19%) of the 37 children had evidence of infection with more than one CMV strain. In six of these children, reinfection with distinct strains was confirmed by analysis of the MIE gene products of sequential CMV strains. CONCLUSIONS: Children who attend child care centers, like adults who are immunosuppressed or have multiple sexual partners, are at risk of being reinfected with distinct CMV strains.