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Biomedical subjects

C H King

Publications and source records attributed to C H King.

At least 19 recordsLinked to original sources
PubMed1999

An in vitro tissue culture bilayer model to examine early events in Mycobacterium tuberculosis infection.

A tissue culture bilayer system that mimics some aspects of early alveolar infection by Mycobacterium tuberculosis was developed. This model incorporates human lung epithelial type II pneumocyte (A549) (upper chamber) and endothelial cell (lower chamber) layers separated by a microporous membrane. This construction makes it possible to observe and quantify the passage of bacteria through the two layers, to observe the interaction of the bacteria with the various cell types, and to examine the basic mechanisms of immune cell recruitment to the site of infection. After 10(7) organisms were added to the upper chamber we microscopically observed large numbers of bacteria attached to and within the pneumocytes and we determined by viable-cell counting that a small percentage of the inoculum (0.02 to 0.43%) passed through the bilayer into the lower chamber. When peripheral blood mononuclear cells were added to the lower chamber, microscopic examination indicated a migration of the mononuclear cells through the bilayer to the apical surface, where they were seen associated with the mycobacteria on the pneumocytes. The added complexity of the bilayer system offers an opportunity to define more precisely the roles of the various lung cell types in the pathogenesis of early tuberculosis.

Cell Culture Techniques↗
PubMed1997

In search of virulence factors of human bacterial disease.

Traditional genetic techniques and a variety of animal and tissue-culture model systems have sustained the study of bacterial virulence mechanisms for several decades. However, the recent application of newly developed molecular and cellular techniques has brought our understanding of bacterial pathogenesis to new heights by permitting the identification and analysis of previously unknown constitutively and differentially expressed virulence-associated factors.

Bacteria↗
PubMed1996

Intracellular growth and cytotoxicity of Mycobacterium haemophilum in a human epithelial cell line (Hec-1-B).

We developed an in vitro model to study the temperature-regulated cytotoxicity and intracellular growth of Mycobacterium haemophilum in cultured human epithelial and endothelial cells. M. haemophilum associated with human epithelial and endothelial cells at similar rates when incubated at 33 and 37 degrees C, but only the epithelial cell line supported the multiplication of this organism. M. haemophilum grew equally well with epithelial cells at both temperatures. The aminoglycoside antibiotic amikacin was used to study the intracellular growth of M. haemophilum in the epithelial cells at 33 and 37 degrees C. Although an approximately equal number of bacteria were found within cells after 2 days of incubation at both temperatures, intracellular replication of M. haemophilum was 1,000-fold greater at 33 than at 37 degrees C. This intracellular multiplication was associated with destruction of the monolayers at 33 but not at 37 degrees C, and only culture filtrates from infected monolayers incubated at 33 degrees C were cytotoxic to fresh epithelial cell monolayers. This strain of M. haemophilum also produced contact-dependent hemolysis of sheep erythrocytes, demonstrating the possible presence of a cytolysin. These studies suggest that M. haemophilum has a preference for growth with cultured human epithelial cells. In addition, intracellular growth is best at 33 degrees C in epithelial cells, and this correlated with cytotoxicity at this temperature. This phenotype may be caused by induction of a soluble cytotoxic component, possibly a hemolytic cytolysin.

Colony Count, Microbial↗
PubMed1996

Comparison of in vitro models for the study of Mycobacterium tuberculosis invasion and intracellular replication.

We recently evaluated several tissue culture model systems for the study of invasion and intracellular multiplication of Mycobacterium tuberculosis. These model systems include a human alveolar pneumocyte epithelial cell line, a murine macrophage cell line (J774), and fresh human peripheral blood-derived macrophages. Our data indicated that the initial level of association of M. tuberculosis with human alveolar pneumocyte cells (2%) was less than that observed with fresh human peripheral blood macrophages (9%) or J774 murine macrophages (13%) within 6 h of the addition of the bacteria. M. tuberculosis replicated in association with the pneumocyte cells by more than 55-fold by day 7 postinfection. In contrast, total bacteria] growth in the J774 cells and human macrophages was considerably less, with increases of only fourfold and threefold, respectively, over the same 7-day period. Amikacin, an aminoglycoside antimicrobial agent, was added to inhibit the growth of extracellular bacteria after the initial 6-h infection period. Decreases in viable counts were observed in all three cell cultures within the first 3 days after infection. However, unlike the case with either macrophage culture, intracellular bacterial CFU obtained from the infected pneumocytes increased by fourfold by day 7 after the addition of amikacin. These data indicate that M. tuberculosis infects and multiplies intracellularly in human lung epithelial cells and that these cells may be an alternative in vitro model for the study of intracellular multiplication of M. tuberculosis in the human lung.

Animals↗
PubMed1996

Dynamics and control of Schistosoma haematobium transmission in Kenya: an overview of the Msambweni Project.

Ecologic modeling of Schistosoma transmission in endemic communities has suggested that antiparasite therapy targeted at the most heavily infected segment of the human population (i.e., school-age children) should have a significant impact on local parasite transmission. Our 1984-1991 experience with age-targeted therapy in the Msambweni area of Kenya has shown an overall decrease in area transmission within 1-2 years following initiation of annual treatment of school-age groups. Snail monitoring confirmed a continuing but variable reduction of vector infection rates. However, subgroup analysis showed significant differences in transmission suppression between more developed coastal villages with piped-water kiosks and villages with only limited access to safe water supplies. Villages without piped water were marked by higher initial prevalences of S. haematobium infection, greater prevalence among adults, longer and more frequent contact with high-risk water sources, and persistently high transmission despite compliance with parasitologic screening or drug therapy. We conclude that targeted therapy had a significant impact on S. haematobium transmission in some areas, but that more extensive or more prolonged coverage is necessary to reduce the rate of new infection in high-risk villages. Defining field-use algorithms, based on decision analysis of economic and ecologic parameters, should provide effective guidelines for selective versus mass treatment in expanded control areas.

Adolescent↗
PubMed1996

Gender differences in growth of school-aged children with schistosomiasis and geohelminth infection.

Light or moderate intensity infection with Schistosoma mansoni may contribute to growth deficits. We report on the effects of treatment for S. mansoni on growth and development in Brazilian schoolchildren. Anthropometric measurements were taken from 539 S. mansoni-infected children and their age- and sex-matched egg-negative controls between the ages of 7 and 15 years. The children as a whole exhibited chronic malnutrition, with growth retardation in height evident in 21% of the population. Infected children, however, were significantly smaller in height, weight, mid upper arm circumference (UAC), tricep skinfold (TSF), and subscapular skinfold (SSF) measurements than control children (P < 0.05). These differences were due primarily to a greater disparity between infected and egg-negative girls in height (P < 0.01), weight (P = 0.01), UAC (P = 0.O2), and TSF (P < 0.01). Nevertheless, girls demonstrated a better level of development and nutrition compared with boys. While infected boys were shorter and weighed less than controls, these differences were not significant. Growth and development in girls was negatively correlated with intensity of infection. Coinfection with S. mansoni and Trichuris appeared to act synergistically in the development of malnutrition.

Adolescent↗
PubMed1995

Molecular biology, virulence, and pathogenicity of mycobacteria.

The diseases resulting from infections with Mycobacterium species are important sources of morbidity and mortality throughout the world today, with particularly devastating effects in tropical and developing countries. Almost 2 billion people have been infected with Mycobacterium tuberculosis, the causative agent of tuberculosis, and approximately 3 million people die each year from this disease. Tuberculosis also has re-emerged as an important public health problem in the United States, and this resurgence has been accompanied by an increased incidence of tuberculosis resistant to the standardly used anti-tuberculosis drugs. Researchers' ability to investigate the molecular basis of the pathogenicity and drug resistance of the mycobacteria has been hampered by a lack of appropriate experimental tools. However, during the past 5 years, tremendous progress has been made in the development of the molecular biology of mycobacteria, and molecular tools are now available for detailed analysis of their genetics and for elucidation of the molecular mechanisms of their pathogenicity. The development of these tools is briefly reviewed, and the uses of the tools to investigate drug resistance in Mycobacterium tuberculosis, to identify mycobacterial virulence factors, and to explore intracellular survival strategies are described.

DNA, Bacterial↗
PubMed1993

Expression of contact-dependent cytolytic activity by Mycobacterium tuberculosis and isolation of the genomic locus that encodes the activity.

We investigated the presence of cytolytic activity in the virulent H37Rv and attenuated H37Ra strains of Mycobacterium tuberculosis and in the vaccine strain Mycobacterium bovis BCG. The virulent strain H37Rv expressed > 3-fold more contact-dependent cytolytic activity than the attenuated strain H37Ra, and the vaccine strain M. bovis BCG did not produce cytolytic activity. We also isolated an approximately 3.2-kbp fragment of the M. tuberculosis H37Rv genome that was capable of inducing this contact-dependent hemolytic activity in a nonhemolytic strain of Escherichia coli.

Animals↗