Boosting survival rates.
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Biomedical subjects
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The national Forest Health Monitoring (FHM) program requires protocols for monitoring soil carbon contents. In a pilot study, 30 FHM plots loblolly shortleaf (Pinus taeda L./Pinus echinata Mill.) pine forests across Georgia were sampled by horizon and by depth increments. For total soil carbon, approximately 40% of the variance was between plots, 40% between subplots and 20% within subplots. Results by depth differed from those obtained by horizon primarily due to the rapid changes in carbon content from the top to the bottom of the A horizon. Published soil survey information overestimated bulk densities for these forest sites. The measurement of forest floor depths as a substitute to sampling did not provide reliable estimates of forest floor carbon. Precision of replicate samples was approximately 10-30% for field duplicates and 5-10% for laboratory duplicates. Based on national indicator evaluation criteria, sampling by depth using bulk density core samplers has been recommended for national implementation. Additional procedures are needed when sampling organic soils or soils with a high percentage of large rock fragments.
Intestinal parasites contribute greatly to morbidity in developing countries. While there have been several studies of the problem in the Caribbean, including the implementation of control programmes, this has not been done for Guyana. The aim of this study was to determine the prevalence of intestinal parasites among young children in a town located in the interior of Guyana. Eighty-five children under the age of 12 years were studied prospectively for intestinal parasites in Mahdia, Guyana. Stool samples were transported in formalin to the Department of Microbiology, The University of the West Indies, Jamaica, for analysis using the formalin-ether concentration and Ziehl-Neelsen techniques. Data on age and gender of the children were recorded on field data sheets. At least one intestinal parasite was detected in 43.5% (37/85) of the children studied and multiple parasitic infections were recorded in 21.2% (18/85). The most common intestinal helminth parasite was hookworm (28.2%; 24/85), followed by Ascaris lumbricoides (18.8%; 16/85) and then Trichuris trichuria (14.1%; 12/85). Among the protozoan infections Giardia lamblia was detected in 10.5% (9/85) of the study population while Entamoeba histolytica appeared rarely. All stool samples were negative for Cryptosporidium and other intestinal Coccidia. There was no predilection for gender with any of the parasites. The pattern of distribution of worms in this area of Guyana was unlike that seen in other studies. Hookworm infection was the most common among the children and a large proportion had multiple infections. The study established the occurrence and prevalence of a number of intestinal parasites in the population of Guyana. This sets the stage for the design and implementation of more detailed epidemiological studies.
Phospholemman (PLM) is a small transmembrane cardiac protein that is the major sarcolemmal substrate for phosphorylation in response to adrenergic stimulation. PLM likely plays a role in muscle contractility and cell volume regulation through its function as a channel or a channel regulator. We are the first to describe the structure of the PLM gene and to demonstrate PLM cDNA splice variants. We cloned the murine PLM cDNA and used it as a probe to isolate the gene from a 129/SvJ genomic library. The gene contains seven introns and eight exons. The coding sequence is interrupted by five introns; the 5' untranslated region by two. Using rapid amplification of 5' cDNA ends we identified transcription start sites and four splice variants of the 5' untranslated domain. There was no TATA box or CAAT box in the putative promoter regions. The gene has several stretches of dinucleotide repeats. The 3' untranslated domains of mouse PLM cDNA clones show sequence differences not accounted for by alternative splicing. Mouse PLM shares 93, 83 and 80% amino acid identity with rat, dog, and human PLMs, respectively. Tissue expression of murine PLM parallels that in other species, being highest in heart, skeletal muscle, and liver.
Cell-imaging approaches using new laser-based technologies have a wide applicability to thefields of pathology and cell biology. Here, we present the application of several of these techniques, including confocal scanning laser microscopy (CSLM), laser scanning cytometry (LSC), and laser capture microdissection (LCM), to studies of cell signaling by environmental agents in lung disease. Using both cells in culture and lung tissue, we show that these technologies are powerful tools for understanding signal transduction cascades elicited by toxic agents, such as oxidants and asbestosfibers, and their relationship to the development of cell injury and proliferation, responses leading to lung disease and/or repair.
Adult female mosquitoes were collected in Mahdia, Guyana, to determine the incidence of malaria in Anopheles species found during the month of June 2000. Centers for Disease Control miniature white (incandescent) light traps, model 512, and miniature black (ultraviolet) light traps, model 912, were used to capture female mosquitoes. Numbers of mosquitoes collected were compared between white and black light traps and between traps set outside and inside of buildings. Adult female Anopheles mosquitoes were identified and an ELISA dipstick test for Plasmodium vivax and Plasmodium falciparum was performed on each mosquito. An aquasalis, An oswaldoi, and An braziliensis were attracted to white light traps. An triannulatus and An darlingi were collected from black light traps. Approximately the same numbers of all female Anopheles mosquitoes 28/45 (62%) were caught inside buildings as outside. Numbers of female non-anopheles mosquitoes captured in light traps varied between the traps set outside of buildings and inside of buildings with bright light traps collecting 91/122 (75%). A total of 45 Anopheles mosquitoes were captured and 122 non-anopheles species. Of the two known vectors of malaria in Guyana, An darlingi mosquitoes were not infected with P vivax but An aquasalis was found to be a carrier. The findings of this study suggest a need for further surveying and identification of current malaria vectors in Guyana.
Sources of Escherichia coli in a coastal waterway located in Ft. Lauderdale, Fla., were evaluated. The study consisted of an extensive program of field measurements designed to capture spatial and temporal variations in E. coli concentrations as well as experiments conducted under laboratory-controlled conditions. E. coli from environmental samples was enumerated by using a defined substrate technology (Colilert-18). Field sampling tasks included sampling the length of the North Fork to identify the river reach contributing high E. coli levels, autosampler experiments at two locations, and spatially intense sampling efforts at hot spots. Laboratory experiments were designed to simulate tidal conditions within the riverbank soils. The results showed that E. coli entered the river in a large pulse during storm conditions. After the storm, E. coli levels returned to baseline levels and varied in a cyclical pattern which correlated with tidal cycles. The highest concentrations were observed during high tide, whereas the lowest were observed at low tide. This peculiar pattern of E. coli concentrations between storm events was caused by the growth of E. coli within riverbank soils which were subsequently washed in during high tide. Laboratory analysis of soil collected from the riverbanks showed increases of several orders of magnitude in soil E. coli concentrations. The ability of E. coli to multiply in the soil was found to be a function of soil moisture content, presumably due to the ability of E. coli to outcompete predators in relatively dry soil. The importance of soil moisture in regulating the multiplication of E. coli was found to be critical in tidally influenced areas due to periodic wetting and drying of soils in contact with water bodies. Given the potential for growth in such systems, E. coli concentrations can be artificially elevated above that expected from fecal impacts alone. Such results challenge the use of E. coli as a suitable indicator of water quality in tidally influenced areas located within tropical and subtropical environments.
There have been dramatic increases in dengue fever (DF) and dengue haemorrhagic fever in South America. Guyana has reported less than five cases per year for most of the past decade. We evaluated patients in a clinic in Georgetown, Guyana, over 2 days and found evidence of 50 cases of dengue infection.
We researched epidemiologic associations between environmental and demographic factors and prevalence of Helicobacter pylori infection in a suburban Jamaican community. Using a clustered sampling technique, 22 domestic yards enclosing 60 separate households were randomly selected from a local community. All household members (n = 346) were invited to participate following informed consent; the overall compliance rate was 58.9%. A commercial enzyme immunoassay (HMáCAP) was used to detect IgG antibodies raised against H. pylori. Environmental and demographic information was obtained by questionnaire. The seroprevalence of H. pylori was 69.9% (n = 202). Analysis of the independent variables revealed three major components: Component 1 described, collectively, good personal hygiene and sanitation, indoor water supply and absence of straying animals in the peridomestic area; Component 2 included older age, good personal hygiene and large yard size; Component 3 the presence of domestic animals (cats and dogs) and, again, large yard size. These three complexes explained 42.2% of the variability in the data set. Logistic regression showed that Components 2 and 3 were independently associated with H. pylori seropositivity, indicating that a combination of demographic, environmental and zoonotic factors is involved in the spread of H. pylori infections at the tropical community level.
An evaluation of total immunoglobulin E (IgE) and dengue serostatus in 168 subjects from San Andrés Island, Colombia, revealed altered levels of IgE in 89% of the population. IgE levels were higher in patients with a history of dengue or with a current secondary or current primary infection than in subjects with no exposure (P = 0. 01). Dengue infection accounted for 23% of the variation in IgE levels.
Rapid detection of human immunodeficiency virus (HIV) infection can result in improved patient care and/or faster implementation of public health preventive measures. A new rapid test, Determine (Abbott, Abbott Park, Ill.), detects HIV type 1 (HIV-1) and HIV-2 antibodies within 15 min by using 50 microl of serum or plasma. No specialized equipment or ancillary supplies are required, and results are read visually. A positive result is noted by the appearance of a red line. An operational control (red line) indicates proper test performance. We evaluated the Determine rapid HIV detection test with a group of well-characterized serum samples (CD4 counts and viral loads were known) and serum samples from HIV-positive individuals at field sites in Honduras and the Dominican Republic. In the field evaluations, the results obtained by the Determine assay were compared to those obtained by local in-country HIV screening procedures. We evaluated serum from 100 HIV-positive patients and 66 HIV-negative patients. All samples gave the expected results. In a companion study, 42 HIV-positive samples from a Miami, Fla., serum bank were tested by the Determine assay. The samples had been characterized in terms of CD4 counts and viral loads. Fifteen patients had CD4 counts <200 cells/mm(3), while 27 patients had CD4 counts >200 cells/mm(3). Viral loads ranged from 630 to 873,746 log(10) copies/ml. All samples from the Miami serum bank were positive by the Determine test. Combined results from the multicenter studies indicated that the correct results were obtained by the Determine assay for 100% (142 of 142) of the HIV-positive serum samples and 100% (66 of 66) of the HIV-negative serum samples. The Determine test was simple to perform and the results were easy to interpret. The Determine test provides a valuable new method for the rapid identification of HIV-positive individuals, especially in developing countries with limited laboratory infrastructures.
We evaluated two new commercial dengue diagnostic tests, the MRL Diagnostics Dengue Fever Virus IgM Capture ELISA and the PanBio Rapid Immunochromatographic Test, on serum samples collected during a dengue epidemic in Jamaica. The MRL ELISA method correctly identified 98% (78 of 80) of the samples as dengue positive, while the PanBio test identified 100% (80 of 80). Both tests were 100% (20 samples of 20) specific.
Malaria is a global health problem, responsible for nearly 3 million deaths each year, and on the increase worldwide. Improvements in malaria diagnostics should facilitate the identification of individuals infected with the malarial parasites and the treatment of such cases with appropriate drugs. Both traditional and contemporary methods for malaria diagnosis are the subjects of the present review. Traditional diagnosis, based on the examination of Giemsa-stained thick and thin blood smears under a microscope, is inappropriate for many areas because there are insufficient microscopes and/or trained microscopists to read and interpret the slides. Such traditional methods are discussed in the context of parasite quantification. Newer, more advanced malaria diagnostics are now available and the relative merits of methods based on fluorescent microscopy or the detection of nucleic acid (including PCR) are described, including comparisons of costs. Fluorescent microscopy and nucleic-acid techniques both require skills and equipment which are not universally available in many malaria-endemic countries. Recently introduced diagnostic tests based on immuno-assays solve this problem since they are easy to run and interpret, and do not require complex equipment or technical support. They are also rapid (< 10 min/test), cost-effective and at least as sensitive as traditional microscopy.
The development of rapid and specific diagnostic tests to identify individuals infected with malaria is of paramount importance in efforts to control the severe public health impact of this disease. This study evaluated the ability of a newly developed rapid malaria diagnostic test, OptiMAL (Flow Inc., Portland, Oreg.), to detect Plasmodium vivax and Plasmodium falciparum malaria during an outbreak in Honduras. OptiMAL is a rapid (10-min) malaria detection test which utilizes a dipstick coated with monoclonal antibodies against the intracellular metabolic enzyme parasite lactate dehydrogenase (pLDH). Differentiation of malaria parasites is based on antigenic differences between the pLDH isoforms. Since pLDH is produced only by live Plasmodium parasites, this test has the ability to differentiate live from dead organisms. Results from the OptiMAL test were compared to those obtained by reading 100 fields of traditional Giemsa-stained thick-smear blood films. Whole-blood samples were obtained from 202 patients suspected of having malaria. A total of 96 samples (48%) were positive by blood films, while 91 (45%) were positive by the OptiMAL test. The blood films indicated that 82% (79 of 96) of the patients were positive for P. vivax and 18% (17 of 96) were infected with P. falciparum. The OptiMAL test showed that 81% (74 of 91) were positive for P. vivax and 19% (17 of 91) were positive for P. falciparum. These results demonstrated that the OptiMAL test had sensitivities of 94 and 88% and specificities of 100 and 99%, respectively, when compared to traditional blood films for the detection of P. vivax and P. falciparum malaria. Blood samples not identified by OptiMAL as malaria positive normally contained parasites at concentrations of less than 100/microl of blood. Samples found to contain P. falciparum were further tested by two other commercially available rapid malaria diagnostic tests, ParaSight-F (Becton Dickinson, Cockeysville, Md.) and ICT Malaria P.f. (ICT Diagnostics, Sydney, Australia), both of which detect only P. falciparum. Only 11 of the 17 (65%) P. falciparum-positive blood samples were identified by the ICT and ParaSight-F tests. Thus, OptiMAL correctly identified P. falciparum malaria parasites in patient blood samples more often than did the other two commercially available diagnostic tests and showed an excellent correlation with traditional blood films in the identification of both P. vivax malaria and P. falciparum malaria. We conclude that the OptiMAL test is an effective tool for the rapid diagnosis of malaria.
We report on our investigation of a malaria outbreak in Honduras, Central America, in January 1997. We tested 202 patients with fever and chills using thin and thick blood film microscopy. Sixteen patients lived in the city and the rest lived in rural areas. A total of 95 samples (47%) were positive for malaria parasites. Seventy-nine percent (63/80) of the rural patients were infected with Plasmodium vivax and 21% (17/80) were infected with P. falciparum. In the urban area, all 15 infected patients had P. vivax malaria and none showed evidence of P. falciparum. Since previous reports indicate that falciparum malaria accounts for only 2% of the overall malaria infections in Honduras, the results reported here suggest that there is a dramatic increase in falciparum malaria in the area of Honduras investigated in this study.
During June 1996, water supplies of the city of San Pedro Sula, Honduras, were sampled to obtain an assessment of Cryptosporidium oocyst and Giardia cyst concentrations. Each sample was concentrated and stained with an indirect immunofluorescent antibody, and parasites were counted through microscopic analysis. In three surface water supplies, Cryptosporidium oocyst concentrations ranged from 58 to 260 oocysts per 100 L, and Giardia cysts were present in concentrations ranging from 380 to 2100 cysts per 100 L. Unlike the surface water samples, groundwater had a higher concentration of Cryptosporidium oocysts (26/100 L) than Giardia cysts (6/100 L), suggesting that the groundwater aquifer protects the water supply more effectively from larger Giardia cysts. Cryptosporidium oocyst concentrations are within the typical range for surface water supplies in North America whereas Giardia cyst concentrations are elevated. Efforts should be made to protect raw water from sources of contamination.
Honduras has at least five-times more human immunodeficiency virus (HIV)-infected individuals than any other country in Central America. The relationship between HIV status and the presence of intestinal parasites in this part of the world is unknown. This study presents the results from a prospective, comparative study for the presence of parasites in 52 HIV-positive and 48 HIV-negative persons in San Pedro Sula, Honduras. Infection with HIV was determined by microagglutination and confirmed by Western blot analysis. Parasites were detected in stools using formalin-ether concentration, and Kinyoun and trichrome staining. Age, sex, and clinical state of HIV infection were recorded for each study participant. Our results indicate that Cryptosporidium parvum and Strongyloides stercoralis, which are intracellular or live in the mucosa, were found exclusively in persons infected with HIV. In comparison, the prevalence of the extracellular parasites Giardia lamblia, Ascaris lumbricoides, and Trichuris trichiura was significantly higher (P < 0.05) in persons who were HIV-negative. Trichuris worms are in contact with the gut epithelium and less so with the mucosa, whereas Strongyloides lives within the gut mucosa. It is possible that changes in the gut epithelium due to HIV infection do not affect the mucosa and therefore would not affect Strongyloides. We conclude that infection with HIV may selectively deter the establishment of certain intestinal parasites. This may be due to the fact that HIV-induced enteropathy does not favor the establishment of extracellular parasites. Intracellular and mucosal dwelling organisms, however, may benefit from pathologic changes and reduced local immune responses induced by the virus, which, in turn, may lead to higher prevalence among HIV-infected individuals.
We report the findings of a cross-sectional epidemiologic study of Giardia lamblia and Cryptosporidum infections in Jamaica. Three hundred twenty eight stool samples from patients less than one to 81 years of age were examined using formalin-ether concentration for G. lamblia, Zeihl-Neelsen staining for Cryptosporidium, and the Prospect rapid enzyme immunoassay (EIA; Alexon, Sunnyvale, CA) for parasite diagnosis. The Prospect Giardia rapid assay detected 17 cases of G. lamblia infection compared with six by formalin-ether concentration. However, the Prospect Cryptosporidum EIA did not increase the rate of detection of Cryptosporidum when compared with Zeihl-Neelsen staining. Cryptosporidum infections were most frequently diagnosed in children less than five years old and prevalence decreased with age. In contrast, the prevalence of giardiasis increased as children became older. There were no associations between the infections and stool consistency, clinical manifestations, or sex of the individuals. The contribution of the parasites to childhood morbidity will depend on accurate laboratory diagnosis.