Biomedical subjects
A Richman
Publications and source records attributed to A Richman.
A modular chitin-binding protease associated with hemocytes and hemolymph in the mosquito Anopheles gambiae.
Sp22D, a modular serine protease encompassing chitin binding, low density lipoprotein receptor, and scavenger receptor cysteine-rich domains, was identified by molecular cloning in the malaria vector, Anopheles gambiae. It is expressed in multiple body parts and during much of development, most intensely in hemocytes. The protein appears to be posttranslationally modified. Its integral, putatively glycosylated form is secreted in the hemolymph, whereas a smaller form potentially generated by proteolytic processing is associated with the tissues. Bacterial challenge or wounding result in low-level RNA induction, but the protein does not bind to bacteria, nor is its processing affected by infection. However, Sp22D binds to chitin with high affinity and undergoes transient changes in processing during pupal to adult metamorphosis; it may respond to exposure to naked chitin during tissue remodeling or damage.
Cloning and analysis of a cecropin gene from the malaria vector mosquito, Anopheles gambiae.
Parasites of the genus Plasmodium are transmitted to mammalian hosts by anopheline mosquitoes. Within the insect vector, parasite growth and development are potentially limited by antimicrobial defence molecules. Here, we describe the isolation of cDNA and genomic clones encoding a cecropin antibacterial peptide from the malaria vector mosquito Anopheles gambiae. The locus was mapped to polytene division 1C of the X chromosome. Cecropin RNA was induced by infection with bacteria and Plasmodium. RNA levels varied in different body parts of the adult mosquito. During development, cecropin expression was limited to the early pupal stage. The peptide was purified from both adult mosquitoes and cell culture supernatants. Anopheles gambiae synthetic cecropins displayed activity against Gram-negative and Gram-positive bacteria, filamentous fungi and yeasts.
Evolution of balanced genetic polymorphism.
Extreme genetic polymorphism maintained by balancing selection (so called because many alleles are maintained in a balance by a mechanism of rare allele advantage) is intimately associated with the important task of self/non-self-discrimination. Widely disparate self-recognition systems of plants, animals and fungi share several general features, including the maintenance of large numbers of alleles at relatively even frequency, and persistence of this variation over very long time periods. Because the evolutionary dynamics of balanced polymorphism are very different from those of neutral genetic variation, data on balanced polymorphism have been used as a novel source for inference of the history of populations. This review highlights the unique evolutionary properties of balanced genetic polymorphism, and the use of theoretical understanding in analysis and application of empirical data for inference of population history. However, a second goal of this review is to point out where current theory is incomplete. Recent observations suggest that entirely novel selective forces may act in concert with balancing selection, and these novel forces may be extremely potent in shaping genetic variation at self-recognition loci.
Patterns of hospital costs for depression in general hospital wards and specialized psychiatric settings.
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In situ fiber-optic oxygen consumption measurements from a working mouse heart.
Luminescence-based imaging-fiber oxygen sensors (IFOSs) were utilized for the in situ measurement of oxygen consumption from intact perfused mouse hearts. IFOSs were fabricated using a technically expedient, photoinitiated polymerization reaction whereby an oxygen-sensitive polymer matrix was immobilized in a precise location on an imaging fiber's distal face. The oxygen-sensing layer used in this work comprised a transition metal complex, Ru(Ph2phen)3(2+), entrapped in a gaspermeable photopolymerizable siloxane membrane (PS802). The transduction mechanism was based upon the oxygen collisional quenching of the ruthenium complex luminescence; detection was performed utilizing an epi-fluorescence microscope/charge coupled device imaging system. IFOS measurements from working mouse hearts were validated through concurrent, blind, ex situ blood gas analyzer (BGA) measurements. The BGA and IFOS methodologies were utilized successfully to measure oxygen concentrations in aortic and pulmonary artery perfusates from the working mouse heart before and after isoproterenol administration. Coupled with coronary-flow measurements, these data were used to calculate myocardial oxygen consumption. Regression analysis of measurements of myocardial oxygen consumption showed that there was a strong correlation between the values generated by the BGA sampling and those obtained via in situ IFOS methods. To our knowledge, this research represents the first report of in situ fiber-optic sensor monitoring of oxygen content from the intact, beating mouse heart.
Antimicrobial activity spectrum, cDNA cloning, and mRNA expression of a newly isolated member of the cecropin family from the mosquito vector Aedes aegypti.
An antimicrobial peptide belonging to the cecropin family was isolated from the hemolymph of bacteria-challenged adult Aedes aegypti. This new peptide, named cecropin A, was purified to homogeneity and fully characterized after cDNA cloning. The 34-residue A. aegypti cecropin A is different from the majority of reported insect cecropins in that it is devoid of a tryptophan residue and C-terminal amidation. The importance of these two structural features on the activity spectrum was investigated using a chemically synthesized peptide. A comparison of the antimicrobial activity spectrum of A. aegypti and Drosophila cecropin A showed a lower activity for the mosquito molecule. A. aegypti cecropin mRNA expression was not detected by Northern blot or reverse transcription-polymerase chain reaction analysis in any immature stage of the mosquito, nor in naïve adults, but it was observed in challenged adults 6 h after bacteria inoculation, and it continued over 7-10 days.
Factors associated with occupational exposure and compliance with universal precautions in an urban school district.
Factors associated with occupational exposure and universal precautions (UP) compliance were assessed among employees in one urban school district. Half of the employees surveyed reported responding to bleeding injuries and cleaning blood or other body fluids (e.g., vomit, urine) during the previous school year. Also, 1 in 4 custodians and 1 in 10 teachers/teacher's aides had direct contact with blood or body fluids without protection. In multivariate logistic regression analyses, direct contact was most likely among secondary school employees in unpredictable situations who did not have protective equipment or comply with UP. UP compliance was greater among those who had protective equipment available and felt self-confident. Self-confidence was associated with having received training or protective equipment. Routine communications between administrators and employees, staff training, provision of protective equipment, and exposure incident monitoring are essential to effective implementation of UP policies in schools and work settings where occupational exposure could occur.
39K NMR measurement of intracellular potassium during ischemia in the perfused guinea pig heart.
The hyperfine shift reagent, TmDOTP5-, was used to resolve the 39K NMR resonances of intra- (Ki+) and extracellular (Ke+) potassium in isolated, perfused guinea pig hearts. [Ki+] as measured by 39K NMR was 25.9 +/- 10.3 mM, compared with 114.4 +/- 10.8 mM as measured by atomic absorption spectroscopy (AAS) using TmDOTP5- as a marker of extracellular space. Thus, only approximately 23% of intracellular potassium was detected by 39K NMR using our experimental conditions. The area of the Ki+ signal increased during early ischemia then returned to baseline levels during reperfusion. In an effort to learn more about the Ki+ not detected by 39K NMR, hearts were perfused with a Rb+-enriched, K+-depleted buffer for an extended period. This resulted in loss of the entire 39K NMR signal, and Ki+, as measured by AAS, decreased from approximately 60 to approximately 6 to 7 micromol/g wet weight. When K+-depleted hearts were subjected to global ischemia, a small 39K NMR signal reappeared, suggesting that at least a portion of the nonexchangeable Ki+ becomes detectable by NMR during ischemia. This newly visible K+ signal subsequently dissipated during reperfusion of ischemic hearts. We conclude that ischemia induces changes in the NMR visibility of 39K in perfused guinea pig hearts.
Molecular immune responses of the mosquito Anopheles gambiae to bacteria and malaria parasites.
Immune responses of the malaria vector mosquito Anopheles gambiae were monitored systematically by the induced expression of five RNA markers after infection challenge. One newly isolated marker encodes a homologue of the moth Gram-negative bacteria-binding protein (GNBP), and another corresponds to a serine protease-like molecule. Additional previously described markers that respond to immune challenge encode the antimicrobial peptide defensin, a putative galactose lectin, and a putative serine protease. Specificity of the immune responses was indicated by differing temporal patterns of induction of specific markers in bacteria-challenged larvae and adults, and by variations in the effectiveness of different microorganisms and their components for marker induction in an immune-responsive cell line. The markers exhibit spatially distinct patterns of expression in the adult female mosquito. Two of them are highly expressed in different regions of the midgut, one in the anterior and the other in the posterior midgut. Marker induction indicates a significant role of the midgut in insect innate immunity. Immune responses to the penetration of the midgut epithelium by a malaria parasite occur both within the midgut itself and elsewhere in the body, suggesting an immune-related signaling process.
Molecular cloning and functional expression of an insect high-affinity Na+-dependent glutamate transporter.
Excitatory amino acid transporters in the central and peripheral nervous systems of insects are thought to assist in maintaining glutamate concentrations in the resting synapse below the activation threshold of glutamate receptors. We have isolated a cDNA from the caterpillar Trichoplusia ni which encodes a high-affinity Na+-dependent glutamate transporter, designated TrnEAAT1. The deduced amino acid sequence shows strong identity with known members of the vertebrate Na+- and K+-dependent amino acid transporter family. Expression of the insect transporter mRNA was predominantly localized in the caterpillar brain. The function of the TrnEAAT1 protein was analyzed in cultured insect cells using a baculovirus expression system. Cells infected with the recombinant virus were found to exhibit a 50-fold increase in ability to accumulate labeled L-glutamate compared to mock-infected cultures, and this activity was shown to be Na+-dependent. Transport activity was further demonstrated by chromatographic identification of various glutamate analogues accumulated by infected cells. Various glutamate uptake inhibitors were used to outline the pharmacological properties of the cloned transporter and to compare it with known mammalian transporters. Despite the significant differences between insect and vertebrate physiology, the characteristics of the respective transporters were found to be remarkably similar.
Identification and characterization of differentially expressed cDNAs of the vector mosquito, Anopheles gambiae.
The isolation and study of Anopheles gambiae genes that are differentially expressed in development, notably in tissues associated with the maturation and transmission of the malaria parasite, is important for the elucidation of basic molecular mechanisms underlying vector-parasite interactions. We have used the differential display technique to screen for mRNAs specifically expressed in adult males, females, and midgut tissues of blood-fed and unfed females. We also screened for mRNAs specifically induced upon bacterial infection of larval stage mosquitoes. We have characterized 19 distinct cDNAs, most of which show developmentally regulated expression specificity during the mosquito life cycle. The most interesting are six new sequences that are midgut-specific in the adult, three of which are also modulated by blood-feeding. The gut-specific sequences encode a maltase, a V-ATPase subunit, a GTP binding protein, two different lectins, and a nontrypsin serine protease. The latter sequence is also induced in larvae subjected to bacterial challenge. With the exception of a mitochondrial DNA fragment, the other 18 sequences constitute expressed genomic sequence tags, 4 of which have been mapped cytogenetically.
Immune factor Gambif1, a new rel family member from the human malaria vector, Anopheles gambiae.
A novel rel family member, Gambif1 (gambiae immune factor 1), has been cloned from the human malaria vector, Anopheles gambiae, and shown to be most similar to Drosophila Dorsal and Dif. Gambif1 protein is translocated to the nucleus in fat body cells in response to bacterial challenge, although the mRNA is present at low levels at all developmental stages and is not induced by infection. DNA binding activity to the kappaB-like sites in the A.gambiae Defensin and the Drosophila Diptericin and Cecropin promoters is also induced in larval nuclear extracts following infection. Gambif1 has the ability to bind to kappaB-like sites in vitro. Co-transfection assays in Drosophila mbn-2 cells show that Gambif1 can activate transcription by interacting with the Drosophila Diptericin regulatory elements, but is not functionally equivalent to Dorsal in this assay. Gambif1 protein translocation to the nucleus and the appearance of kappaB-like DNA binding activity can serve as molecular markers of activation of the immune system and open up the possibility of studying the role of defence reactions in determining mosquito susceptibility/refractoriness to malaria infection.
Immunity to eukaryotic parasites in vector insects.
Mosquitoes and blackflies have been the focus of recent efforts to elucidate factors influencing the susceptibility of vector insects to metazoan and protozoan parasites of medical significance. Vector species exhibit variation in cellular and humoral immune responses, as highlighted by studies of melanotic encapsulation and components of the phenoloxidase system. Significant progress has been made in the development of genetic maps based upon molecular markers, leading to the genetic analysis of loci influencing susceptibility. The identification of specific inducible antibacterial peptides, and the cloning of genes encoding immune effector proteins as well as potential regulatory factors, open the path to fruitful studies of vector insect innate immunity and its relationship to insect-parasite interactions.
The 'good mother'--a comparative study of Swedish, Italian and American maternal behavior and goals.
This paper is based on a comparative, psychologically informed ethnographic study of maternal goals and infant care during the first year of life for 20 mothers and infants in each of three Western settings: Stockholm, Sweden, outside Rome, Italy, and suburban Boston, in the United States. Two research questions were posed to consider cultural ideas and ideals about mothering; How was the 'good mother' defined? How did sample women themselves, mother? Research hypotheses were that definitions of a good mother would be culture-specific with minimal within-culture variance. Furthermore, the frequency of maternal behaviors would also exhibit culture specific patterns. Research methods included event based behavioral observations of infant-caregiver interactions in the home, daily routine questionnaires, maternal attitude interviews, and ethnographic observations of community and family life. Research hypothesis were confirmed. Results revealed culture-specific variation in conceptions of the 'good mother'. Cultural differences were reflected in the short- and long-term goals which the sample women described for their children. Patterns of infant care, in turn, were consistent with the cultural values expressed, as well as the cultural norms of social interaction.
Level of care for deinstitutionalized psychiatric patients.
The New York state Level-of-Care Survey (LOCS) was used in 1987-88 to assess the community care and social support needs of 936 mentally disabled community residents in Halifax, Nova Scotia. The cases were assigned an appropriate level-of-care by the computerized algorithm based on an 101-item questionnaire covering all aspects of the cases' physical and mental condition, needs and activities. Compared with their current setting, the algorithm assigned 60% of cases to the same level-of-care, 17% to a lower level, and 23% to a higher level-of-care. The net effect on social service costs of the reassignments would be a 7% increase. The increased costs for some cases ($1.5 million) would be partly offset by lowered costs for others ($900,000). The gradual shifting of 14 cases from the relatively expensive rehabilitation unit to more independent living at lower levels of care would compensate for most of the increases.
National health insurance and private psychiatry.
From 1972-73 to 1988-89, the number of Canadian psychiatrists doubled. Most psychiatrists are in fee-for-service practice. The median earnings of psychiatrists are close to those of internists. There is lack of psychiatric services available, but not a lack of psychiatrists. The lack of services is due largely to the allocation of psychiatric resources. The maldistribution of psychiatrists is increasing as more settle in urban centres. The U.S. General Medical National Advisory Council recommends that psychiatry spend two thirds of clinical time treating severe illnesses such as psychoses. Before any changes are made to the structure of Canadian psychiatry, more data are needed on patterns of fee-for-service practice, the characteristics of patients, the course and outcome of private psychiatric care, and alternative patterns of practice in which psychiatrists are consultants to family physicians and non-medical therapists.
Does psychiatric care by family practitioners reduce the cost of general medical care?
This is the first "impact" type of economic study of psychiatric/counseling services from general practitioners. The paper analyzes a province-wide database that collates statistical data from all inpatient and outpatient psychiatric services as well as from private physicians. This paper asks whether psychiatric services from family physicians also reduce the overall costs of medical care. This research supports the general research findings that medical costs are lower after psychiatrists' care. ECT patients show a marked reduction in their medical costs. Patients with psychotherapy/counseling from family physicians did not show statistically significant reductions in overall medical costs. We need new classifications for the kinds of mental disorders seen in primary care settings.