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Biomedical subjects

R Hoff

Publications and source records attributed to R Hoff.

At least 37 records · Page 2Linked to original sources

Polymerase chain reaction in detecting HIV infection among seropositive infants: relation to clinical status and age and to results of other assays.

The polymerase chain reaction (PCR) for human immunodeficiency virus (HIV) was evaluated using coded blood specimens from infants whose clinical status is now known. A micromethod for the efficient isolation of mononuclear cells from small volumes of blood, and definitions of PCR positivity that took into account the number and purity of these mononuclear cells, were established in an attempt to define parameters for quality assurance. Results of HIV culture, p-24 antigen, and HIV-specific IgA obtained on the same specimens were compared to PCR results. PCR had a specificity of 100% among 83 specimens from 50 babies known to be uninfected. Sensitivity among 26 HIV-infected infants older than 3 months was 98% (44 of 45 specimens); the one negative specimen, which had also been culture negative, gave a positive PCR result on the remaining aliquot when tested after decoding. Among infected infants less than 3 months old, which is an age when diagnosis by other assays is most problematic, PCR identified 10 of 10 patients (10 of 11 specimens) including two younger than one month. Viral culture showed the best concordance with PCR; however, in three infants, positive PCR results were observed several months before positive results were observed by viral culture.

Aging↗

Clinical utility of HIV-IgA immunoblot assay in the early diagnosis of perinatal HIV infection.

OBJECTIVE: To ascertain the sensitivity, specificity, predictive value, and clinical use of a human immunodeficiency virus (HIV)-IgA immunoblot assay for diagnosing perinatal HIV infection in infants tested at birth to 1 month, 3 months, and 6 months of age. DESIGN: Prospective, longitudinal cohort study of children born to HIV-infected and noninfected women. The HIV-IgA immunoblot assays were performed at birth to 1 month, 3 months, and 6 months of age and compared with the Centers for Disease Control's classification system of HIV infection in the children. Children were followed up for at least 15 months to ensure accuracy of infection status. SETTING: Municipal hospital in central Brooklyn, NY, where the prevalence of HIV infection is high. PATIENTS: Serum samples from 58 children, 22 with documented HIV infection, 18 noninfected children born to seropositive women, and 18 children born to noninfected women, were studied. MAIN OUTCOME MEASURE: Diagnosis of HIV infection using the Centers for Disease Control's classification scheme was compared with diagnosis using the HIV-IgA immunoblot assay for children 6 months of age or younger. RESULTS: The HIV-IgA immunoblot assay yielded negative results at 3 and 6 months of age for all 18 infants born to seronegative women; for the 18 seroreverting, noninfected children born to infected women, the assay yielded negative results at 1 month, 3 months, and 6 months of age. The positive predictive value of the assay was 100%--no false-positive results were identified in the 88 serum samples obtained from noninfected infants. For the HIV-infected children, sensitivity was a function of age: one (5.9%) of 17 infants had an assay that yielded positive results at birth to 1 month of age, 13 (62%) of 21 infants had assays that yielded positive results at 3 months of age, and 17 (77%) of 22 infants had assays that yielded positive results at 6 months of age. The presence or absence of symptoms did not affect the sensitivity. CONCLUSION: The HIV-IgA immunoblot assay can detect a significant proportion of infected children during an early asymptomatic period of their life. This relatively inexpensive, easily standardized assay may allow for institution of therapy before the onset of clinical symptoms.

Female↗

Prevalence of HIV infection in childbearing women in the United States. Surveillance using newborn blood samples.

A national, population-based survey was initiated in 1988 to measure the prevalence of human immunodeficiency virus (HIV) infection in women giving birth to infants in the United States. Following standardized procedures, residual dried-blood specimens collected on filter paper for newborn metabolic screening were tested anonymously in state public health laboratories for maternal antibody to HIV. As of September 1990, annual survey data were available from 38 states and the District of Columbia. The highest HIV seroprevalence rates were observed in New York (5.8 per 1000), the District of Columbia (5.5 per 1000), New Jersey (4.9 per 1000), and Florida (4.5 per 1000). Nationwide, an estimated 1.5 per 1000 women giving birth to infants in 1989 were infected with HIV. Assuming a perinatal transmission rate of 30%, we estimate that approximately 1800 newborns acquired HIV infection during one 12-month period. Preventing transmission of HIV infection to women and infants is an urgent public health priority.

Anonymous Testing↗

IgA and IgM human immunodeficiency virus antibodies in weakly reactive or false-negative blood donors.

Detection of antibodies to the human immunodeficiency virus (HIV) in recently infected donors is crucial to prevent the transmission of HIV infection via blood products. To determine whether specific antibodies of the IgA or IgM class are present as markers of recent infection in donor specimens that have borderline reactivity on routine enzyme immunoassay (EIA) screening, 15 specimens that were positive by immunoblot were tested for IgA and IgM HIV antibodies. All 15 had detectable IgA HIV antibodies, and 14 had IgM HIV antibodies. The 15 specimens were tested further, each by two independent laboratories, with nine licensed EIAs. Two of the nine EIAs found all 15 units positive in both laboratories; seven EIAs found 1 to 5 of the 15 units negative, for a total of 31 false-negative results. The results indicated a difference between the sensitivity of EIA kits using only anti-IgG reagents and of kits using multispecific reagents that react with IgG and other classes of antibody. In a modified procedure, the addition of enzyme-conjugated anti-IgA or anti-IgM to the kit's enzyme-conjugated reagent increased the optical density values of most false-negative specimens to the positive range. It was concluded that licensed kits vary in reactivity with IgA and IgM HIV antibodies and that sensitivity could be increased by improved detection of these classes of antibody.

False Negative Reactions↗

Isocratic reversed-phase HPLC method to measure pyrimethamine extracted from plasma of infants treated for toxoplasmosis.

An isocratic HPLC method for measuring pyrimethamine extracted from infant plasma is reported. The method is an improvement over previously published methods by requiring lower volumes of plasma (100 microL) and having increased sensitivity to pyrimethamine at 210 nm. The procedure, which entails a basic organic extraction and subsequent HPLC chromatography of the reconstituted extract, can detect 1.4 ng and quantify 4.0 ng of pyrimethamine per 40-microL injection, with two analyses per 100-microL sample. Analytical recovery of pyrimethamine added to plasma at 10, 50, and 125 ng/100 microL averaged 80%, 92%, and 101%, respectively (n = 20). Within- and between-day CVs were less than 7%. Studies of various plasma samples from adults and infants (n = 15) revealed no interference from other plasma peaks with the analyte of interest.

Chromatography, High Pressure Liquid↗

Detection of IgM and IgA HIV antibodies after removal of IgG with recombinant protein G.

Indirect assays for IgM and IgA antibodies often lack sensitivity and specificity due to interference from IgG antibodies. To overcome this problem we have developed a simple procedure using recombinant protein G coupled to agarose beads to remove the interfering IgG. A series of HIV seroconversion panels was tested by IgM and IgA immunoblot after protein G treatment in order to evaluate IgG removal and to study appearance of IgM and IgA antibodies in early HIV infection. Protein G treatment removed 99.9% of the IgG and reduced IgG anti-HIV titers of over 1/100,000 to undetectable levels. Both IgM and IgA HIV antibodies were detected as early in seroconversion as were IgG HIV antibodies. IgA HIV antibodies persisted for a longer period of time, reacted with more HIV proteins, and showed more intense staining than IgM HIV antibodies.

Bacterial Proteins↗

Gross morphological changes in the salivary glands of Ixodes ricinus (Acari, Ixodidae) between bloodmeals in relation to active uptake of atmospheric water vapour.

The gross morphological changes in the salivary glands of Ixodes ricinus (L.) were investigated at the light microscopic level in various phases off the host with emphasis on the engorged nymph, in order to relate the capability of active vapour uptake in the course of postembryonal development to degeneration and regeneration of salivary-gland alveoli. Agranular alveoli in engorged immatures of I. ricinus, from detachment to the following early pharate phase, do not appear different from those of the unfed instars. This is also true for the female up to approximately the end of oviposition. During moulting, the agranular alveoli of the immatures degenerate and new ones are formed which are apparently already functional in teneral nymphs and adults. In contrast, granular alveoli, much enlarged in freshly detached immature I. ricinus, shrivel in the early post-repletion period and soon reach a highly reduced state which is maintained until apolysis. Subsequently, they disintegrate completely. The finding that engorged and detached immatures of I. ricinus with markedly atrophied granular alveoli are capable of active vapour uptake until some days after initiation of apolysis suggests that only agranular alveoli are responsible for producing the primary secretion involved in vapour uptake.

Animals↗

Use of paper-absorbed fingerstick blood samples for studies of antibody to human immunodeficiency virus type 1 in intravenous drug users.

The suitability of paper-absorbed (PA) fingerstick blood specimens for antibody testing of human immunodeficiency virus type 1 (HIV-1) was examined in two populations of intravenous drug users (IVDU): 393 persons from a drop-in counseling and testing clinic and 145 from a methadone treatment clinic. From the first group, the same 66 immunoblot-confirmed enzyme immunoassay (EIA)-positive specimens were identified in sera from venipuncture and parallel fingerstick PA specimens. The latter had slightly higher EIA mean background levels resulting in 10 immunoblot-negative EIA-positive samples versus 6 in the sera group. HIV-1 seroprevalence was 17% of 393 from the drop-in clinic. By category of IVDU, the rates were 34% and 14% for active and recovering IVDU, respectively (P less than .001), and 36% in black and Latino compared with 13% in white IVDU (P less than .002). Of the 145 participants in the methadone program, 39% had antibody to HIV-1: 49% for blacks and Latinos compared with 30% in whites (P less than .01). The data indicate that antibody testing for HIV-1 by PA is equivalent to the serum antibody assay of venipuncture specimens. The fingerstick method appears to have greater use for serosurveys and screening programs because of convenience, safety, and ease of storage, transport, and processing of samples.

AIDS Serodiagnosis↗

A nine year prospective study of Chagas' disease in a defined rural population in northeast Brazil.

The association between infection with Trypanosoma cruzi and the development of electrocardiographic alterations and mortality was studied longitudinally for 9 years in a population residing in an endemic area in Castro Alves, Bahia, Brazil. Annual censuses were begun in 1973. At 3 year intervals from 1974 to 1983, the population was examined serologically and subjected to electrocardiography using standardized techniques. Of 1,751 individuals registered during the 10 censuses, 1,541 (88%) were examined serologically at least once. Of 747 individuals in constant residence and originally seronegative, 24 were seropositive upon subsequent evaluation, giving a seroconversion rate of 4.92/1,000 person-years (PY). The overall rate of development of an abnormal ECG was 25.7/1,000 PY for seropositive individuals and 12.5/1,000 PY for seronegative individuals, a relative risk of 2. The 10-14-year-old seropositive group had the highest risk of developing ECG abnormalities (24.1/1,000 PY, relative risk = 3.5). The age-adjusted mortality rate of seropositive individuals was slightly higher than for seronegative individuals (8.9 vs. 7.8/1,000 PY). In sero-positive individuals, mortality was strongly associated with the presence of ventricular conduction defects and arrhythmias. Ventricular conduction defects appeared most frequently in younger individuals. Older individuals presented the highest risk of development of frequent and multifocal extrasystoles.

Adolescent↗

Seroprevalence of human immunodeficiency virus among childbearing women. Estimation by testing samples of blood from newborns.

Attempts to predict the course of the epidemic of acquired immunodeficiency syndrome (AIDS) have been hampered by the lack of an objective, practical way to estimate the prevalence of infection with the human immunodeficiency virus (HIV) in the general population. Testing for the prevalence of HIV infection in women should be a sensitive means to track the epidemic and to study the potential for perinatal transmission. Antibodies in maternal blood are contained in neonatal blood specimens routinely collected on absorbent paper for other purposes, such as screening for phenylketonuria; we therefore tested for HIV antibody in these specimens. Analysis of batches of individually blinded specimens from selected hospitals protected the anonymity of the mothers and babies and was cost efficient. Using the newborn's blood as an indicator of the mother's serologic status, we concluded that 1 of every 476 women (2.1 per 1000) giving birth in Massachusetts was positive for HIV antibody by immunofluorescence assay or enzyme-linked immunosorbent assay, both confirmed by immunoblot (Western blot) testing. The prevalence of HIV infection varied according to the type and location of the maternity hospitals; rates of seropositivity were highest in inner-city hospitals (8.0 per 1000), lower in mixed urban and suburban hospitals (2.5 per 1000), and lowest in suburban and rural hospitals (0.9 per 1000). This method is useful for collecting data needed to plan and evaluate prevention strategies and to predict the health care resources that will be needed to care for women and children who contract AIDS. Because other states have newborn screening programs similar to the Massachusetts program, this approach can be used for national surveillance of AIDS in women.

Acquired Immunodeficiency Syndrome↗

Isolation and functional characterization of murine T cell lines and clones specific for the protozoan parasite Trypanosoma cruzi.

Murine T cell lines responsive to the protozoan parasite Trypanosoma cruzi were generated in vitro by stimulating hyperimmune C57BL/6 lymphoid cells with trypomastigote stage antigen. A spleen-derived line designated ST1 and eight clones derived from ST1 were characterized. All lines bear the surface phenotype Thy-1.2+, Ly-1.2+, 2.2- and respond to T. cruzi antigen only in the presence of antigen-presenting cells matched at the I-A subregion of the H2 locus. Clonal specificity analyses indicated that these T. cruzi-selected T cells are species specific and recognize antigenic determinants that are expressed predominantly in the trypomastigote stage. On the basis of their distinct patterns of response to a panel of different T. cruzi strains, clones recognizing strain-specific, shared, or common determinants were identified. Functional studies indicated that ST1 and some but not all of the clones are capable of expressing antigen-specific T helper function in vitro and in vivo. In addition, co-incubation of T. cruzi-specific T cells with cultured T. cruzi-infected syngeneic macrophages led to the dose-dependent destruction of intracellular parasites. Most notably, ST1 and several of the cloned T. cruzi-specific T cell lines were able to passively protect syngeneic recipients from lethal T. cruzi challenge infection. Efforts to identify the parasite antigens recognized by these T cell lines, particularly the protective clones, are currently in progress.

Animals↗

Cardiac morbidity and mortality due to Chagas' disease: prospective electrocardiographic study of a Brazilian community.

The evolution of Chagas' cardiomyopathy is poorly understood. We therefore examined the development of cardiac lesions in a rural Brazilian community for a period of 7 years. Initially, 42% of 1017 residents were seropositive for infection with Trypanosoma cruzi. Age-specific infection rates indicated that most had become infected before the age of 20 years. On follow-up, it appeared that those persons who developed cardiac lesions did so soon after infection, since the incidence of right bundle branch block and other ventricular conduction defects (VCDs) was also highest before age 20 years. The progressive nature of these lesions was demonstrated by frequent development of additional electrocardiographic abnormalities and high mortality among infected adults with VCDs. In contrast, mortality was low and approximately the same for seropositive and seronegative adults under 60 years who had normal electrocardiograms. Electrocardiography during the early asymptomatic stage of infection was able to distinguish persons with potentially lethal cardiac lesions from those with a benign prognosis.

Age Factors↗

Manson's schistosomiasis in Brazil: 11-year evaluation of successful disease control with oxamniquine.

This prospective study has shown that oxamniquine treatment controlled endemic schistosomiasis mansoni in a defined rural population in Castro Alves, north-east Brazil. Data before and after treatment spanning 11 years were collected for a cohort of 191 residents. Before treatment (1974-77), the cohort was heavily infected and the prevalence of associated hepatomegaly (greater than 86%) and splenomegaly (greater than 17%) was stable. The cohort was treated when oxamniquine became available in 1977; during the next 8 years, over 80% received further treatments from the Brazilian programme for the control of schistosomiasis. With treatment, the incidence of splenomegaly fell (10% to 2%) and the splenomegaly regression rate increased (43% to 91%). Declining disease rates were coincident with substantial falls in the prevalence and intensity of Schistosoma mansoni infections. The final prevalence rates for hepatomegaly (31%) and splenomegaly (3%) in Castro Alves approached the corresponding rates of 10% and 1% in a comparable uninfected control population.

Adolescent↗

Heterogeneous distribution of neuraminidase activity in strains and clones of Trypanosoma cruzi and its possible association with parasite myotropism.

Various strains, stocks, and clones of Trypanosoma cruzi were analyzed for neuraminidase (NA) activity using fetuin and human erythrocytes as substrate. In all cases the activity was found to be developmentally regulated. Zymodeme type I strains, which are histotropic for skeletal muscle, had greater NA activity than zymodeme type II strains which are histotropic for either macrophages or cardiac muscle cells. Heterogeneity of NA expression within strains is suggested by the finding that one Silvio X10 clone had greater NA activity than another clone of the same stock. The differences observed were more pronounced when human erythrocytes and not fetuin were used as substrate. Trypomastigotes of the high producing strains reared in bovine artery smooth muscle cells had enhanced expression compared to trypomastigotes reared in 3T3 or human fibroblast cells. The first harvest of trypomastigotes from cell cultures had greater NA activity than trypomastigotes harvested on subsequent days.

Animals↗

An outbreak of Chagas' disease in southwestern Bahia, Brazil.

An outbreak of 20 cases of acute Chagas' disease followed the movement of Triatoma infestans into the county of Riacho de Santana, Bahia, Brazil. The outbreak was unusual in that the majority of cases occurred in adults. Vector control measures were implemented. Three years after the outbreak, a rural community was examined to determine the extent of human infection and disease due to Trypanosoma cruzi. Ninety of 440 residents (20.5%) had serologic evidence of infection, but rates of electrocardiographic (EKG) abnormalities were low. Comparison of age-specific rates of seropositivity and EKG abnormalities with rates from areas with endemic Chagas' disease supported the hypothesis of a recent epidemic. Control measures appear to have interrupted transmission in the region.

Adult↗