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Biomedical subjects

C R Roberts

Publications and source records attributed to C R Roberts.

At least 55 records · Page 3Linked to original sources

Host feeding preference of Phlebotomus guggisbergi, a vector of Leishmania tropica in Kenya.

Recently the sandfly Phlebotomus guggisbergi was found to be a vector of Leishmania tropica causing cutaneous leishmaniasis in the Laikipia focus, Kenya, but extensive searches have shed no light on the identity of the rural reservoir host(s). In order to discover more about the biology of the vector, a host feeding preference study was conducted on wild sandflies in their natural cave environment over a 6-month period. Solid state Army miniature (SSAM) traps, without light bulb, were suspended over cages with potential hosts or an empty cage control. The animals tested included sheep, goat, dog, cat, hamster, rabbit, giant rat (Cricetomys gambianus), crested rat (Lophiomys imhausi) and rock hyrax (Procavia capensis), all of which (except hamsters) are normally found in the vicinity of the study site. Sandfly collections from traps baited with goat, sheep, cat, dog, rabbit, or wild rodent species were significantly higher than the control, whereas trap collections with hamster and rock hyrax were not significantly different from the control. Numbers of sandflies collected from the goat, sheep and cat were significantly greater than from the rabbit and rodents. The sex ratio also varied between collections: larger animals attracted a higher proportion of female P. guggisbergi than did the smaller animals (P > 0.05). Therefore greater emphasis should be placed on surveying larger animals to assess their status as reservoir hosts for L. tropica in Kenya.

Animals↗

Safety and efficacy of human growth hormone treatment in girls with Turner syndrome.

Six major adverse medical events (AMEs) and 44 minor AMEs were recorded in 6 and 23 patients, respectively, during human growth hormone (GH) treatment of Turner syndrome, equivalent to 118 treatment years. During the first year of GH treatment, there was no change in the SD score (SDS) for systolic or diastolic blood pressure. In a subgroup of 20 patients, there was no change in psychological tests, oral glucose tolerance, glycosylated haemoglobin or serum triglycerides over the first year of GH treatment. The mean total serum cholesterol level fell over this period. The low incidence of adverse medical problems during GH treatment complemented its efficacy. Height velocity in the first year was 7.7 +/- 1.8 cm/year (mean +/- SD) and was correlated with maternal height SDS, though not with paternal height SDS. Some 36% of first-year height velocity SDS could be predicted by the dose of GH and maternal height SDS.

Adolescent↗

Leishmania donovani parasites in the nasal secretions, tonsillopharyngeal mucosa, and urine centrifugates of visceral leishmaniasis patients in Kenya.

In the early 1930s, investigators of visceral leishmaniasis stated that Leishman-Donovan bodies are found in body fluids of kala-azar patients, for example, in urine, feces, semen, and nasal and pharyngeal secretions. Based on this finding, we investigated the diagnostic potential of nasal secretions, tonsillopharyngeal mucosal swabs, and urine centrifugates inoculated into Schneider's Drosophila Medium (containing antibiotics and antifungal agents) as well as with Giemsa-stained smears. Consequently, 64 randomly selected patients with visceral leishmaniasis from Kenya (59 who were splenic culture or Giemsa stain positive and five who were culture negative but Giemsa stain positive) were tested by three noninvasive methods. These tests were all performed before the patients were treated with Pentostam. Cultures of nasal and tonsillopharyngeal swabs and urine centrifugates produced 28 positive samples representing 24 patients (37.5%). Moreover, a set of 25 Giemsa-stained slide smears made from the nasal and tonsillopharyngeal mucosa of 25 patients with visceral leishmaniasis who had not tested positive in cultures produced nine positives. Therefore, the overall total of patients who tested positive by all of the above methods was 33 or 51.6%. The cryopreserved Leishmania isolates were characterized by cellulose acetate electrophoresis using 20 enzyme systems. The isoenzyme profiles produced by the parasites were represented in five different L. donovani s.l. zymodemes. Representatives of these isolates were also characterized by DNA Southern blotting analysis, which corroborated the isoenzyme results.

Adolescent↗

Factors affecting exflagellation of in vitro-cultivated Plasmodium falciparum gametocytes.

The environment of Plasmodium falciparum gametocytes changes when they make the transition from the vertebrate to the invertebrate host. Gametocytes of this species cultivated in vitro were used to evaluate the effect of serum, pH, pCO2 tension, bicarbonate ion, and temperature on gamete formation. Temperature was the only factor responsible for keeping P. falciparum gametocytes in the inactivated state. Mature gametocytes held at temperatures above 30 degrees C remained quiescent in 10% serum, even at low ambient pCO2 tension, alkaline pH, and in the presence of 25 mM bicarbonate ion. When the temperature of the medium was allowed to drop below 30 degrees C, gametocytes emerged from the red blood cells and microgametocytes consistently exflagellated at pH 7.4, even in the absence of bicarbonate ion. With regard to bicarbonate ion, exflagellation in P. falciparum is similar to P. berghei and different from P. gallinaceum gametocytes, which have an obligate requirement for bicarbonate ion.

Animals↗

Cultivation of Plasmodium falciparum parasites in a serum-free medium.

The elimination of serum from Plasmodium falciparum culture media could decrease costs, enhance procurement, and improve the feasibility of large-scale production of parasite material. We provide a semi-defined, serum-free formulation, of commercially available constituents that supports P. falciparum parasite growth at rates comparable with those obtained with serum-supplemented media. The medium is composed of RPMI 1640 to which HEPES, extra glucose, bicarbonate, and hypoxanthine have been added. Bovine albumin and serum-derived, lipids-cholesterol-rich mixture are then used in place of serum.

Animals↗

Three-year incidence study of retroviral and viral hepatitis transmission in a Peruvian prostitute population.

A Peruvian female prostitute population was evaluated over a 3-year period to determine the incidence and risk factors of retroviral and viral hepatitis transmission. At three survey periods, a questionnaire was administered and serum samples were obtained. A total of 966 subjects were studied, with 34% followed for 38 months, 22% followed for 18 months, and 44% evaluated just once. On initial evaluation, 3 (0.3%) had HIV-1 antibody, 170 (17.6%) had HTLV-I antibody, 578 (59.8%) had anti-HBc, and 7 (0.7%) had antibody to hepatitis C virus. The mean annual incidence of HTLV-I and hepatitis B infection was 1.6% and 4.7%, respectively. Univariate and logistic regression analysis of prevalence data indicated an association between sexual activity and HTLV-I and hepatitis B infection, but no independent risk factors were identified in cohort analysis. Parenteral risk factors were not associated with transmission, except for a small percentage of subjects who may have acquired hepatitis B infection from blood transfusions. These findings suggest that there is a high incidence of HTLV-I and hepatitis B infection from heterosexual contact in this female prostitute population.

Adult↗

Working mothers and infant care: a review of the literature.

1. The number of women who returned to work after childbirth increased steadily through the 1980s, with many mothers returning to work earlier than ever before. This trend has caused new mothers, their employers, and legislators to examine the issue of infant day care. 2. Infant day care availability, arrangements, and cost influence maternal work force participation. These concerns may also impact on the physical and mental health of the working mother. 3. Further research is needed to determine if and how infant day care concerns affect worker productivity, and to determine how to obtain quality day care at a reasonable cost.

Child Day Care Centers↗

Comparative evaluation of six rapid serological tests for HIV-1 antibody.

Six commercial rapid test kits for HIV-1 antibody were evaluated. Four laboratory technologists tested sera from four groups of U.S. military personnel or dependents: 202 subjects positive for HIV-1 antibody by Western blot, 200 seronegative voluntary blood donors, 199 seronegative obstetrics/gynecology patients, and 99 subjects with sera reactive by ELISA but negative by recombinant protein EIA and indeterminate by Western blot. The three tests using solid-phase immunoassay technology demonstrated the highest mean sensitivity (> 99%) and specificity (> 91%) for all groups tested, including sera indeterminate by Western blot. Two dot-immunoblot assays were less specific, possibly due to indistinct reaction end points, and a latex agglutination assay was also less specific because of difficulty distinguishing reactive results from the granular background. In an "ease-of-use" assessment, solid-phase capture immunoassays required less time and equipment and were easier to interpret than other testing methods. Solid-phase capture immunoassays for HIV-1 antibody may be suitable for use in emergency situations and in developing countries because they are highly sensitive and specific and are rapidly performed with minimal laboratory equipment.

AIDS Serodiagnosis↗

Effect of controlled burning on survival of floodwater Aedes eggs in Kenya.

The effect of controlled burning on the survival of Aedes mosquito eggs was evaluated in 2 distinct dambo habitats. In a dambo dominated by grasses, egg survival was 3.3% after burning compared with 43.8% in a similar dambo that was not burned. In a dambo dominated by sedges, egg survival was 0.7% after burning compared with 28.5% in a similar dambo that was not burned. Mortality of mosquito eggs appeared to be caused by high temperatures associated with the fire and not elapsed time since egg survival did not decrease with time after burning. The potential for burning to control the mosquito vectors of Rift Valley fever virus is discussed.

Aedes↗

Failure to detect evidence of human T-lymphotropic virus (HTLV) type I and type II in blood donors with isolated gag antibodies to HTLV-I/II.

Of the 267,650 blood donations from members of the US armed forces, 72 (0.027%) were serologically confirmed to be positive for human T-lymphotropic virus type I/II (HTLVpos) and 379 (0.14%) were Western blot (WB)-indeterminate with banding pattern restricted to the proteins encoded by the gag gene only (HTLVind). To determine whether these apparently healthy HTLVind blood donors are infected with HTLV-I or HTLV-II, coded specimens from randomly selected military blood donors (n = 73) were tested for antibodies to HTLV by WB and radioimmunoprecipitation assay (RIPA) using HTLV-I (MT-2) antigens, by enzyme immunoassay using synthetic peptides representing the immunodominant epitopes of HTLV, and for sequences of proviral HTLV DNA by the polymerase chain reaction (PCR). Of the 73 HTLVind donors, none showed presence of env reactivity by HTLV WB and RIPA. Minimal reactivity was observed with synthetic immunodominant motifs derived from the env protein of HTLV-I (Env-1(191-214) and Env-5(242-257)) or HTLV-II (Env-2(187-209) and Env-20(85-102)) and gag protein (Gag-1a(102-117) and Gag-10(364-385)). A peptide corresponding to the endogenous retroviral sequence with structural homologies to the gag protein of HTLVs (RTVLgag) reacted with antibodies not only in HTLVpos (88%) and HTLVind (42% to 66%) specimens, but also reacted with normal control subjects (60%). Furthermore, none of the 73 HTLVind specimens demonstrated presence of the HTLV genome when amplified with primers for the pol and tax/rex region. Six to 23 months from the initial test, 27 subjects still gave indeterminate WB patterns, and 13 of these repeat specimens were still negative for the presence of HTLV genome. We conclude that individuals at low risk for HTLV infection who have HTLVind WB reactivity are rarely, if ever, infected with HTLV-I or HTLV-II.

Amino Acid Sequence↗

Serologic differentiation of human T lymphotropic virus type I from type II infection by synthetic peptide immunoassays.

Synthetic peptide-based serologic assays (Select-HTLV and SynthEIA) that distinguish the closely related human T-cell lymphotropic virus types I (HTLV-I) and II (HTLV-II) were tested blindly for their ability to correctly identify infection caused by either virus type. Of 57 HTLV-I and 38 HTLV-II specimens from individuals whose infections were confirmed by polymerase chain reaction, the Select-HTLV assay categorized 56 (98%) as HTLV-I, 36 (95%) as HTLV-II, and 3 (3%) as nontypeable. Similarly, the SynthEIA assay categorized 54 (95%) as HTLV-I, 29 (76%) as HTLV-II, and 12 (13%) as nontypeable. More importantly, no specimen was wrongly classified by either assay (100% specificity). Further, analysis of serial specimens from six patients also demonstrated concordant results with the PCR findings. Our results suggest that serotyping of HTLV infections can be achieved reliably by these peptide assays, thus the need for technically complex PCR-based HTLV typing, while not eliminated, can be greatly reduced.

Base Sequence↗

Sexual transmission of human T-lymphotropic virus type I in Peruvian prostitutes.

The epidemiology of HTLV-I infection in female prostitutes was studied in a survey of 395 prostitutes from Callao, Peru (the port city of Lima), 72 prostitutes from Iquitos, Peru (another port city on the Amazon River), and 510 prenatal clinic patients from Lima. Prostitutes reported a mean of 8.8 years (range, 1-39 years) of active prostitution and a mean of 205 sexual contacts during the month prior to the study. The percentage of prostitutes with HTLV-I antibody (21.8%) was significantly higher than patients attending a prenatal clinic (3.1%; P less than .0001). The prevalence of HTLV-I antibody increased steadily with age in prostitutes, but no age trend was noted in prenatal patients. By multiple logistic regression analysis, an independent association was found between HTLV-I seropositivity and a history of prostitution in Callao, age, and positive syphilis serology when all 977 study subjects were evaluated. When prostitutes alone were analyzed, the number of years of exposure as a practicing prostitute was associated with HTLV-I seropositivity after controlling for age. These data indicate a greatly increased risk of HTLV-I infection in prostitutes in Callao, Peru, and suggest an association between sexual activity and HTLV-I transmission.

Adolescent↗

Immune responsiveness to the immunodominant recombinant envelope epitopes of human T lymphotropic virus types I and II in diverse geographic populations.

The heterogeneity of immune responsiveness to the immunodominant epitopes of human T lymphotropic virus (HTLV) types I (MTA-1(162-209)) and II (K-55(162-205)) were determined in natural infections with HTLV-I and -II from diverse geographic areas (n = 285). Of the HTLV-I specimens confirmed by polymerase chain reaction (PCR), all North American (n = 37) and Peruvian (n = 19) specimens reacted with MTA-1. Of HTLV-II specimens confirmed by PCR, 44 (96%) of 46 from North American blood donors, 28 (97%) of 29 from native Americans, and all from intravenous drug users (n = 29) reacted with K-55. Specimens from other geographic areas (Peru, 30; Brazil, 4; Mexico, 10; Italy, 5; Somalia, 13; Ethiopia, 17; Japan, 32; and Jamaica, 15) all reacted either with MTA-1 or K-55. By synthetic peptide-based serologic typing, all of these specimens could be typed as HTLV-I or -II. In addition to the direct implications of these findings for diagnostic purposes, these data provide indirect evidence for the conservation of immunodominant HTLVenv epitopes in diverse geographic populations.

Africa↗

Sensitivity of a ribosomal RNA gene probe for identification of life stages of Anopheles arabiensis and An. gambiae (Diptera: Culicidae) using three storage methods.

Individual larvae, pupae, female adults, and adult body parts of Anopheles arabiensis Patton and An. gambiae Giles were stored for 1 mo either in isopropanol at room temperature, over a desiccant at room temperature, or at -70 degrees C. DNA was extracted, digested with EcoR1 restriction enzyme, subjected to electrophoresis in agarose gel, transferred to filters, then hybridized to a 32P-labeled rDNA probe. There was no difference among storage treatments in the proportion of correctly identified samples. First instars were not identifiable. Pupae and female adults were more likely to be identified than earlier life history stages. Nonetheless, the probe identified greater than 75% of second instars, 94% of third instars, and 74% of fourth instars. There were no differences between the species in the proportion of identifiable samples for any life history stage.

Animals↗

Mechanical properties of human tracheal cartilage.

Biomechanical changes in airway cartilage could influence the mechanics of maximal expiratory flow and cough and the degree of shortening of activated airway smooth muscle. We examined the tensile stiffness of small samples of human tracheal cartilage rings in specimens obtained at autopsy from 10 individuals who ranged in age from 17 to 81 yr. The tensile properties of the cartilage were compared with its content of water (%water), glycosaminoglycans (chondroitin sulfate equivalents, mg/mg dry wt), and hydroxyproline content (mg hydroxyproline/mg dry weight). The average values for tensile stiffness ranged between 1 and 15 MPa and increased significantly with increasing age [tensile stiffness = 0.19 x (age in yr) + 2.02; r = 0.83, P less than 0.05]. The outermost layer of cartilage was the most stiff in all individuals, and the deeper layers were progressively less stiff. Water content and hydroxyproline content both decreased with increasing age. Thus tensile stiffness correlated inversely with water content and hydroxyproline content [tensile stiffness = -0.83 x (%water) + 16.4; r = 0.82, P less than .05 and tensile stiffness = -342 x (hydroxyproline content) + 25; r = 0.87, P less than 0.05]. Total tissue content of glycosaminoglycans did not change with age, although changes in glycosaminoglycan type and proteoglycan structure with increasing age have been described. We conclude that there are age-related changes in the biomechanical properties and biochemical composition of airway cartilage that could influence airway dynamics.

Adolescent↗

Prevalence of human T-lymphotropic virus in civilian applicants for the United States Armed Forces.

BACKGROUND: The wide range in human T-lymphotropic virus (HTLV) seroprevalences reported worldwide has made estimates of seroprevalence difficult in unique populations. In this study, the seropositivity of young adult civilian applicants for the US Armed Forces was determined. METHODS: Serum samples from nine geographic regions were screened by an Enzyme-Linked Immunosorbent Assay (ELISA), and repeatedly reactive samples were further tested by Western blot and radioimmunoprecipitation. Specimens were scored as positive when antibody to gag (p24) and env (gp46 or gp68) were detected. RESULTS: Of the 43,750 samples analyzed, 18 were positive for HTLV antibodies. Ten (55%) were from males and eight (45%) were from females. Nine (90%) of the males and seven (87.5%) of the females were Black. Twelve of the positive samples (66.6%) were from the New York City region, which represented only 18.8% of the sample population. CONCLUSIONS: The overall HTLV seroprevalence of civilian applicants for the US Armed forces was 0.41 per 1000. This was higher than the seroprevalence reported for volunteer blood donors.

Adolescent↗

Biochemical characterization and zymodeme classification of Leishmania isolates from patients, vectors, and reservoir hosts in Kenya.

A total of 407 Leishmania and other Leishmania-like isolates obtained from patients, other vertebrates, sand fly vectors, and other arthropods from Kenya and other countries were characterized and compared with several World Health Organization and other well-characterized reference strains of Leishmania, Trypanosoma, Crithidia, Herpetomonas, and Leptomonas by cellulose acetate electrophoresis (CAE), using 20 enzyme systems. Analysis of the isoenzyme banding patterns (IBP) of the isolates generated isoenzyme profiles that were resolved as zymodemes and tabulated. Isolates that produced similar isoenzyme profiles in all 20 enzyme systems were placed into a particular Leishmania isoenzyme taxon, with the zymodeme designated numerically as Zn. A total of 66 zymodemes were recorded for the 407 isolates studied. To obviate the need to draw all 66 representative IBP for each of the 20 enzyme systems, the 66 zymodemes (Z1-Z66) were again placed into similarity groups represented by pattern number or Pn. This resulted in 23-50 IBP (Pn) per enzyme system. The highest number of IBP scored was for malate dehydrogenase (MDH) (P1-50) and the lowest score was for glucose-6-phosphate isomerase (GPI) (P1-23). From these different isoenzyme profiles or zymodemes, IBP of 14 (MDH, GPI, nucleoside hydrolase, phosphoglucomutase, malic enzyme, isocitrate dehydrogenase, glucose-6-phosphate dehydrogenase, mannose-6-phosphate isomerase, 6-phosphogluconate dehydrogenase, glutamate oxaloacetate transferase/aspartate aminotransferase, glutathione reductase, superoxide dismutase, fumarase, and glyceraldehyde-3-phosphate dehydrogenase) of the 20 enzyme systems were selected for computer-calculated numerical taxonomy. Consistent individual isoenzyme bands with similar relative mobilities of the 14 enzyme systems were scored into groups (allelomorphs, allozymes, or electromorphs) and used in cluster analysis. For each pattern in every profile, the presence of a consistent band was entered as 1 and its absence as 0. A total of 419 allozyme characters (variables) were scored for the 14 enzyme systems. Lastly, all different zymodemes sharing a particular IBP (Pn) within an enzyme system were counted and the total number was shown as a zymodeme frequency (Zf). Final analysis of the CAE isoenzyme profiles and cluster-dendrograms resulted in the identification of several potentially new species and subspecies of Leishmania and other Leishmania-like isolates from patients, sand flies, and animal reservoir hosts collected from Kenya and other locations in Africa. Zymodeme analysis of the Kenyan visceral and cutaneous leishmaniasis isolates resulted in the identification of 11 subpopulations of the L. donovani species complex and six subpopulations of the L. tropica species complex endemic to different geographic areas of Kenya.

Animals↗

Characterization of HIV isolates arising after prolonged zidovudine therapy.

Human immunodeficiency virus type 1 (HIV-1) was isolated from five patients with late-stage disease treated with zidovudine (ZDV) for more than 1 year. Peripheral blood mononuclear cells (PBMCs) were used for all virus isolations and to assay for drug resistance. The isolates exhibited a 10- to 100-fold decrease in ZDV susceptibility compared to pretreatment isolates. Multiple clones of a 618 bp segment of the HIV reverse transcriptase gene encompassing codons 60-250 were sequenced for each isolate. The association of alterations at codons Asp67----Asn, Lys70----Arg, Thr215----Phe or Tyr, and Lys219----Gln with ZDV resistance has been previously noted (ref. 5). In this study, the most frequent alterations was Thr215----Tyr although genotypic mixtures of Thr/Tyr and Phe/Tyr were also observed. One isolate with a Tyr215 alteration and unaltered codons at 67, 70, and 219 had high-level ZDV resistance. Alterations at codons 67, 70, and 219 did not appear to increase resistance when seen in combination with Tyr215. Virus isolates obtained from each patient by cultivation with either 0 or 4 microM ZDV were compared and found to have similar alterations at codons 67, 70, 215, and 219, although one instance of apparent in vitro selection for Tyr215 over Phe215 was observed. Assays using PBMCs for virus propagation will permit susceptibility testing of HIV isolates from most patients on antiretroviral drugs to investigate the clinical significance of drug resistance.

Amino Acid Sequence↗