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Biomedical subjects

R D Robinson

Publications and source records attributed to R D Robinson.

At least 37 records · Page 2Linked to original sources

Classification of red blood cells as normal, sickle, or other abnormal, using a single image analysis feature.

Sickle cell anemia is a disease for which there is currently no effective treatment. One method of evaluating clinical status is the counting of cell types based on morphology. There is a need for a rapid, reproducible method, superior to human inspection, for classification of these cells. Quantitative digital-image analysis is being applied to this need. Blood from 24 patients with sickle cell anemia (SS) and SC disease and ten hematologically normal volunteers (AA) was stressed by bubbling with nitrogen. One hundred fifty cells were analyzed from each sickle specimen, and 100 were analyzed from each nonsickle specimen. Expert observers classified each cell as normal (N), sickle (S), or other abnormal (A). Cells were analyzed with a custom, high-resolution image-analysis instrument. A total of 42 features including metric, optical density-derived, and textural features were extracted. The metric feature Form Factor (4 pi Area/Perimeter2) was selected by recursive partitioning analysis as the sole feature needed for segregating cells into the classes of N, A, and S. The agreement of automated classification (using cutpoints determined by recursive partitioning analysis) with a human expert for specimens from individuals with sickle cell anemia was 89% for N-, 73% for A-, and 92% for S-classified cells. For specimens from AA individuals, the agreement was 92% for N and 76% for A. For specimens from individuals with sickle cell anemia, rates of agreement between two human experts were compared and found to be 86% for N, 84% for A, and 80% for S. For specimens from AA individuals, the agreement was 90% for N and 87% for A.

Adolescent↗

Textural differences between AA and SS blood specimens as detected by image analysis.

High-resolution image analysis was employed in the analysis of round (discoid) erythrocytes from hematologically normal (AA) individuals, AA individuals with nonspecific anemia, individuals with sickle cell trait (AS), individuals with SC disease (SC), and individuals with sickle cell anemia (SS). The shape feature Form Factor (4 pi Area/Perimeter2) was used to select round cells and to exclude sickle and other abnormal cells. Textural features extracted from round cells of SS and SC patients were found to differ from those derived from cells of normal andanemic AA individuals. Two textural features, Standard Deviation of Run Length Matrix Counts and Rotation Moment of the Cooccurrence Matrix, discriminated between patients mean values from AA samples and those from SS samples. The ability of textural features to separate round cells into classes based on genotype suggests that high resolution image analysis may be an effective tool in the study and monitoring of sickle cell disease.

Adolescent↗

Strongyloides stercoralis: characterization of immunodiagnostic larval antigens.

Forty-one-, 31-, and 28-kDa proteins of Strongyloides stercoralis filariform larvae have previously been demonstrated to be sensitively and specifically recognized by serum IgG in individuals with strongyloidiasis. Characteristics of these proteins, their immunodominant epitopes, and reactive antibodies are described here. The proteins are soluble in aqueous as well as detergent extracts. The immunodominant epitopes are present in S. stercoralis but not in S. cebus or S. ratti. Epitopes on the three proteins are not shared, as determined by cross-adsorption of serum with each of the size components on nitrocellulose. In most sera from strongyloidiasis patients there was reactivity to each of the proteins by IgG1 and IgG4, but reactivity by IgG2 or IgG3 was detectable only in a minority. A rabbit antiserum raised to a 41-kDa size fraction of S. stercoralis larvae reacted against a doublet of 41-kDa which was distinct from the immunodiagnostic 41-kDa protein.

Animals↗

Use of pleural Tenckhoff catheter to palliate malignant pleural effusion.

Malignant pleural effusion and its treatment both cause substantial morbidity in patients with advanced neoplastic disease. We hypothesized that this morbidity might be ameliorated by placement of an indwelling Tenckhoff catheter into the involved pleural space. Catheters were placed in 9 patients under local anesthesia. Three patients underwent bilateral catheter placement, for a total of 12 catheters placed. Four of the 9 patients had undergone previous unsuccessful pleurodesis (using tetracycline or bleomycin). Whenever it became symptomatic, the malignant pleural effusion was simply drained into a calibrated container and the volume recorded. Patients were followed on a weekly basis until their death (mean, 16 weeks). The mean drainage was 477 mL per 24 hours (range, 200 to 1,100 mL). No pleural space infections occurred, although local cellulitis developed in 3 patients around the catheter exit site; all patients responded to oral antibiotics. There were no significant changes in either the serum albumin or total protein levels. No catheters malfunctioned and no patients required further treatment or hospitalization for symptoms of malignant pleural effusion. We conclude that this technique may reduce the morbidity stemming from malignant pleural effusion and its treatment by allowing patients to conveniently and painlessly drain the effusion at home when it becomes symptomatic. This technique may provide superior palliation in patients with malignant pleural effusion.

Adult↗

Ocular pharmacokinetics of orally administered azithromycin in rabbits.

Azithromycin was orally administered to Dutch-belted rabbits following extracapsular lens extraction in one eye. At various times the animals were sacrificed, and serum and ocular tissues were obtained for drug level determination by HPLC-EC. Following a single dose, peak levels of drug in ocular tissues were measured within 8 hours (cornea > 0.5 micrograms/g [15mg/kg]; > 1.5 micrograms/g [3Omg/kg]). Highest levels were obtained in iris and ciliary body ( > 15 micrograms). Measurable tissue levels persisted for at least 120 hours. Trough levels increased proportionately during drug multiple dose administration. Five days following five daily 15mg/kg doses, corneal levels exceeded 0.5 micrograms/g, and iris and ciliary levels were higher than 15 micrograms/g. Aqueous humor and serum levels were equivalent. Vitreous humor levels, though higher than aqueous humor, were consistently < 1 microgram/ml. Extracapsular cataract extraction did not significantly affect drug uptake.

Administration, Oral↗

Immunoepidemiologic studies of Strongyloides stercoralis and human T lymphotropic virus type I infections in Jamaica.

Epidemiologic investigations of Strongyloides stercoralis and human T lymphotropic virus type I (HTLV-I) infections were conducted. Of 312 persons contacted, 209 (67%) provided blood and stool samples. Prevalences of S. stercoralis and HTLV-I antibodies were 26.8% and 8.1% (n = 198), respectively, and S. stercoralis larvae were detected in 4%. HTLV-I antibodies were significantly more common in persons positive for S. stercoralis larvae (10 [58.8%] of 17) compared with seropositive larva-negative (4 [8.9%] of 45) or seronegative persons (9 [6.2%] of 145) (P < .002). IgE levels increased with age in S. stercoralis-seropositive persons who were HTLV-I negative (P < .002). However, there was an age-related depression of serum IgE in HTLV-I-positive positive persons (P < .003) that was sufficient to annul the IgE level-raising effect of S. stercoralis seropositivity. The data provide evidence that HTLV-I infection is associated with increased frequency of larvae in the stool of S. stercoralis-infected persons and suggest that the mechanism may involve suppression of the IgE response.

Adult↗

Prospective evaluation of enzyme-linked immunosorbent assay and immunoblot methods for the diagnosis of endemic Strongyloides stercoralis infection.

Recently described enzyme-linked immunosorbent assay (ELISA) and immunoblot methods for the detection of serum IgG against Strongyloides stercoralis larval antigens were prospectively evaluated for the diagnosis of endemic strongyloidiasis. A modification of the ELISA involved preincubation of sera with Onchocerca gutturosa phosphate-buffered saline-soluble extract to remove cross-reactivity with other helminths. The sensitivity of the ELISA increased from 80% to 85% following preincubation. Similarly, there was an increase in specificity from 94% to 97%. The IgG recognition of 41-, 31-, and 28-kD filariform larval components showed sensitivities of 100%, 85%, and 65%, respectively. Both the ELISA following incubation of sera with O. gutturosa extract and serum IgG reactivity to a 41-kD larval component using immunoblotting are sensitive and specific techniques for diagnosing endemic strongyloidiasis.

Animals↗

Endoreplication in the ovary, testis, and intestine of Strongyloides stercoralis.

During development of the free-living adults of the human parasitic nematode Stronglyoides stercoralis, cells in certain tissues grow by endoreplication in which rounds of DNA replication occur without cell or nuclear division. The DNA content of individual nuclei was measured by microdensitometry of Feulgen-stained preparations. In females, some ovarian cells have up to 800 times the haploid DNA content (800C). In males, some cells of the testis have up to 100C. Intestinal cells in both sexes have up to 16C, whereas most other somatic cells have 2C.

Animals↗

Chromosome complement, gametogenesis, and development of Strongyloides stercoralis.

Gametogenesis and development were studied in free-living adults of the human parasitic nematode Strongyloides stercoralis. The diploid chromosome number is 6 in germ-line tissue of females and in embryos that will develop into parasitic females. Reproduction appears to be by meiotic parthenogenesis and pseudogamy, as in other species in the genus. Fecundity may be limited by the short lifespan of males. Newly hatched larvae contain about 500 cells, whereas adult females have about 840 somatic cells and a variable number of germ-line cells. The apical vegetative zones of both ovary and testis are occupied by cells with large amounts of DNA in their nuclei.

Anaphase↗

Why are some pamphlets hard for patients to read?

In this study with lung cancer patients, we describe not only text characteristics that hinder comprehension, including unfamiliar words and poor organization, but also reader characteristics that hinder comprehension, such as educational level, and what the patient wants to know. Based on our and other's research we recommend writing what the reader wants to know, relating new learning to what the learner already knows, and using organizational cues for clarity and emphasis.

Aged↗

Serum IgG reactivity with 41-, 31-, and 28-kDa larval proteins of Strongyloides stercoralis in individuals with strongyloidiasis.

Proteins from a deoxycholate-soluble extract of Strongyloides stercoralis infective larvae were separated by SDS-PAGE, blotted onto nitrocellulose paper, and reacted with sera from individuals with confirmed S. stercoralis infections (n = 100), suspected S. stercoralis infections in whom no larvae could be detected (n = 27), and other nematode infections (40 with Wuchereria bancrofti, 20 with Onchocerca volvulus, 20 with Necator americanus, and 20 with mixed Ascaris lumbricoides and Trichuris trichiura infections). Immunodominant proteins of approximately 41, 31, and 28 kDa were recognized by IgG in 91%, 88%, and 90%, respectively, of sera from those with confirmed strongyloidiasis; in 100%, 100%, and 93% of sera from those with suspected strongyloidiasis; and in 9%, 12%, and 14% of sera from those infected with other nematodes. IgG reactivity to each of these proteins was a more specific means of immunodiagnosis than the currently used indirect ELISA; the methods were equally sensitive.

Animals↗

The qualitative evaluation of the pharmacokinetics of subconjunctivally injected antifungal agents in rabbits.

Dutch-belted rabbits with corneal epithelium either intact or debrided were injected subconjunctivally with 300 microliters of one of six antifungal agents: 10 mg/ml miconazole, 5 mg/ml fluconazole, 5 mg/ml ketoconazole, 2.5 mg/ml itraconazole, and 5 mg/ml amphotericin B. At intervals of 10 min to 96 h after injection, animals were killed and corneas removed at the limbus. Three vertical strips from the right cornea and four contiguous 3-mm disks trephined from the central vertical axis of the left cornea were placed on agar plates seeded with an appropriate indicator organism. After 24 h of incubation, the zones of inhibition were measured. For itraconazole, miconazole, fluconazole, saperconazole, and ketoconazole, central corneal levels peaked by 2 h in normal and debrided corneas. Little or no drug was detectable after 4-8 h, except for itraconazole, which persisted in the cornea for at least 24 h in both normal and debrided corneas. Peak levels of amphotericin B in the central cornea were achieved after 2 h in rabbits with debrided corneas, with no drug activity measured after 8 h. There were no detectable levels of drug found in the central corneas of rabbits with intact corneal epithelium. On the basis of this pilot study, the method offers a rapid approach to the screening of antifungal agents for possible use by subconjunctival injection.

Animals↗

Ocular pharmacokinetics of saperconazole in rabbits. A potential agent against keratomycoses.

The ocular pharmacokinetics of saperconazole, an experimental lipophilic triazole with activity against filamentous fungi, including Aspergillus and Candida species, were evaluated in rabbits by radioassay. The drug was administered by topical, subconjunctival, and oral routes. Following a single 20-microL drop of 0.25% saperconazole in normal corneas, a mean (+/- SEM) peak level of 2.32 +/- 0.06 micrograms/g was achieved in 10 minutes. In débrided corneas, a peak level of 13.09 +/- 2.87 micrograms/g was achieved in 2 minutes. The drug was rapidly cleared from the cornea within 2 hours. The administration of 13 drops during 1 hour resulted in a threefold increase in normal corneal levels and in a sixfold increase in débrided cornea levels. Peak levels following subconjunctival injection in normal corneas (12.91 +/- 2.02 micrograms/g) were approximately twofold greater than those following sustained topical administration (6.19 +/- 0.16 micrograms/g) and, in débrided corneas, were a third higher than those following topical therapy in débrided corneas. Clearance was virtually complete by 8 hours. Levels following oral administration were low and probably subtherapeutic in all ocular tissues that were evaluated. Bioassay studies revealed that 44.17% of the drug in the cornea following topical administration was bioactive.

Administration, Oral↗

The evaluation of therapeutic responses in experimental keratomycosis.

Two different measures of response to therapy were evaluated in a model of keratitis caused by Aspergillus fumigatus in Dutch-belted rabbits. Combined pre and post-inoculation treatment with oral fluconazole 37.5 mg/kg bid or itraconazole 40 mg/kg bid was compared to post-inoculation treatment only and untreated controls using a standardized clinical disease severity score and quantitative isolate recovery techniques. For both drugs, there was no difference in isolate recovery rates among all three groups. However, a significant improvement in clinical disease was noted in the pre and post-inoculation treatment group compared to controls (p less than .01) and to the post-inoculation group (p less than .05) for fluconazole. A similar trend, though not statistically significant, was apparent with itraconazole treatment. This disparity highlights the difficulties associated with measuring responses to therapy in keratomycosis and emphasizes the need for more sensitive and specific measures.

Administration, Oral↗

Anomalous effect of subconjunctival miconazole on Candida albicans keratitis in rabbits.

After intrastromal injection of a standardized inoculum of Candida albicans blastoconidia in Dutch-belted rabbits, the efficacy of subconjunctival therapy with amphotericin B and miconazole was evaluated using a quantitative isolate recovery technique. The subconjunctival injection of miconazole resulted in the recovery of significantly more viable organisms compared to the number recovered in controls (2,000 micrograms daily, P less than .02; 3,000 micrograms daily, P less than .01, respectively). Amphotericin B, in contrast, in dosages of 500 or 1,500 micrograms daily, had a significant therapeutic effect (P less than .03). The anomalous response to subconjunctival therapy with miconazole seen in this model suggests an effect on host defense mechanisms.

Amphotericin B↗

Influence of corticosteroid on experimentally induced keratomycosis.

To assess the effect of corticosteroid on the establishment of experimentally induced keratomycoses, rabbits were injected subconjunctivally with triamcinolone acetonide on two successive days before inoculation with Candida albicans, Aspergillus fumigatus, or Fusarium solanae. Whereas isolate recovery rates declined steadily in normal control corneas, they remained stable over 15 days in corticosteroid-treated corneas. Clinically, inflammation was equivalent (A fumigatus and F solanae) or significantly less (C albicans; P = .001) until the 10th day. At 15 days, inflammation in corticosteroid-treated corneas was significantly worse in animals infected with A fumigatus (P = .003) or F solanae (P = .02). Inflammatory signs correlated inconsistently with isolate recovery. Pathogenicity of the infecting organism appears to be important in determining the degree to which corticosteroid is able to mask clinical signs of infection while enhancing fungal replication.

Animals↗