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Biomedical subjects

H B Tanowitz

Publications and source records attributed to H B Tanowitz.

At least 55 records · Page 3Linked to original sources

Utility of microsporidian rRNA in diagnosis and phylogeny: a review.

This paper summarizes work done in this laboratory over the last two years on the cloning of microsporidian rRNA by homology PCR and its subsequent use in diagnostic tests and phylogenetic studies. Using highly conserved primers in the 16S or small subunit rRNA (SSU-rRNA) these genes were cloned from human intestinal biopsies with transmission electron microscopy proven Enterocytozoon bieneusi and Septata intestinalis. The SSU-rRNA genes were then used to design and test several primer pairs for the diagnosis of microsporidian infection. Utilizing the polymerase chain reaction and primers V1 and EB450 Ent. bieneusi infected duodenal aspirates or intestinal biopsies could be detected. Using V1 and SI500 infection with S. intestinalis could be detected. In addition to diagnostic tests, phylogenetic relationships were examined using sequence data from the fragment amplified by PCR by primer 530f in the SSU-rRNA and primer 580r in the large subunit rRNA. This data supported the placement of S. intestinalis in the family Encephalitozoonidae. In addition, it confirmed that Encephalitozoon cuniculi, E. hellem and S. intestinalis are distinct organisms. These techniques have broad applications to the study of other microsporidia and the development of a molecular phylogeny.

Animals↗

Diagnosis and treatment of common intestinal helminths. II: Common intestinal nematodes.

In the second article of this series, we discuss the diagnosis and treatment of some important intestinal nematodes. Common parasites discussed in detail include Ascaris lumbricoides, Trichuris trichiura, hookworm, and Enterobius vermicularis (pinworm). In addition, Strongyloides stercoralis has emerged as an important pathogen in immunocompromised patients, and Capillaria philippinensis is now recognized as an important cause of diarrhea in Asia. Finally, anisakiasis and other illnesses due to parasitic nematodes are discussed.

Adult↗

Evidence that myocardial pertussis toxin substrates are uniquely altered in acute murine Chagas' disease in a manner unrelated to myocardial dysfunction.

In an effort to correlate biochemical characteristics of the beta-adrenergic receptor complex with myocardial function, mouse myocardial GTP-binding proteins, specifically substrates for pertussis toxin (PT), were analysed with regard to the influence of infection with Trypanosoma cruzi, the causative agent of Chagas' cardiomyopathy. Infection was found to decrease in a non-uniform manner the magnitude of ADP-ribosylation in the PT substrates. High detergent concentrations attenuated the infection-associated decrease in PT-dependent ADP-ribosylation. Infection also altered the kinetics of the PT-dependent ADP-ribosylation reaction from a time course wherein maximal PT-dependent ADP-ribosylation occurred after 12 h incubation in control animals to one in which maximal PT-dependent ADP-ribosylation occurred after 3 h incubation and thereafter declined. Immunochemical analysis of the PT-substrates revealed an infection-associated decrease in alpha i1, alpha o, an increase in alpha i2 and no change in alpha i3. Verapamil treatment, which prevents the clinical consequences of infection, did not influence any of the infection-associated changes in PT-dependent ADP-ribosylation of GTP-binding protein substrates or their immunochemical properties. Complementary studies using isolated rat neonatal cardiocytes infected with the parasite further substantiated the finding that the infection-associated decrease in PT-dependent ADP-ribosylation and the associated change in the kinetics of the reaction were properties uniquely associated with the presence of the parasite.

Acute Disease↗

Efficacy of azithromycin for treating Babesia microti infection in the hamster model.

Because of its prevalence and severity, Babesia microti infection is an important public health problem. The current treatment of choice is clindamycin plus quinine. However, in some cases other treatments are needed because of drug intolerance or relapse. The activity of azithromycin was investigated for treatment of babesiosis in the hamster model. All animals received vancomycin to prevent antibiotic-associated colitis. Quinine (250 mg/kg/day), azithromycin (150 mg/kg/day), and the combination of azithromycin and quinine were compared. A significant suppression of parasitemia was found in all treatment groups (combination had the greatest effect, followed by azithromycin, then quinine; P < .05). The mean survival was significantly prolonged in the combination group (P < .05). Azithromycin as monotherapy in a higher dose (300 mg/kg/day) also resulted in a significant prolongation of survival (P < .05). Spirogermanium and ciprofloxacin, which have been reported to have antimalarial activity, had no effect on parasitemia or survival in this experimental babesiosis model.

Animals↗

Small subunit rRNA sequence of Enterocytozoon bieneusi and its potential diagnostic role with use of the polymerase chain reaction.

In the past several years, microsporidia have become recognized as another important group of opportunistic infections of immunocompromised patients, especially those with AIDS. Enteric infections with the noncultivatable microsporidian parasite Enterocytozoon bieneusi have been diagnosed from AIDS patients with chronic diarrhea, malabsorption, and wasting. The incidence of infection and mechanism of transmission of these organisms in humans is unknown. Several recent tests for human pathogens have been developed using rRNA genes as diagnostic probes. Using the polymerase chain reaction and conserved regions of the small subunit rRNA (SSU-rRNA) gene, the SSU-rRNA gene of E. bieneusi was successfully cloned and subsequently sequenced. Amplification of E. bieneusi rRNA could be demonstrated from intestinal biopsies from HIV-1-infected patients infected with E. bieneusi but not from intestinal biopsies from noninfected patients. This cloned SSU-rRNA gene was used to develop improved probes for detection of E. bieneusi in tissue of infected patients.

AIDS-Related Opportunistic Infections↗

Diagnosis and treatment of intestinal helminths. I. Common intestinal cestodes.

Increase in travel and immigration has led to a heightened awareness of parasitic diseases among health professionals. Intestinal helminths are important human parasites. Cestodes or tapeworms comprise an important group of helminths. The diagnosis of tapeworm infections requires a skilled laboratory because serological tests are unavailable. In this first part of our review of intestinal helminths, we describe the salient features of the diagnosis and treatment of several important intestinal cestodes, including Diphyllobothrium latum, Taenia, Hymenolepis, and Dipylidium caninum. Niclosamide and praziquantel are the drugs of choice for tapeworm infections.

Adult↗

Parasitic infections in AIDS patients. Cryptosporidiosis, isosporiasis, microsporidiosis, cyclosporiasis.

AIDS is characteristically associated with several intracellular enteric protozoan infections that often cause chronic and sometimes fatal intractable large-volume diarrhea. Until the AIDS epidemic, several of these parasitic infections were almost unknown as causes of human disease. This article reviews the diseases produced by cryptosporidia, isospora, cyclospora, and microsporidia in humans.

AIDS-Related Opportunistic Infections↗

Taeniasis and cysticercosis.

This article focuses on clinical issues of taeniasis and cysticercosis, including a comprehensive review of the clinical data, standard and latest chemotherapy, modern concepts of pathogenesis, conventional and advanced diagnostic tests, current epidemiology, and effective means of control. Fundamental parasitology is covered to familiarize physicians and scientists with the latest concepts of parasites.

Adrenal Cortex Hormones↗

Trypanosoma cruzi: stage expression of calmodulin-binding proteins.

The subcellular distribution of calmodulin-binding proteins in three life stages of Trypanosoma cruzi was analyzed by a [125I]calmodulin gel overlay procedure under conditions where proteolysis was kept to a minimum. It was found that T. cruzi contains a complex profile of calcium-dependent calmodulin-binding proteins and that several of these polypeptides were differentially expressed at specific stages of development. The majority of these stage-specific polypeptides was found in the particulate fractions of the replicative stages of the parasite, i.e., epimastigote and amastigote. These studies suggest that calcium and calmodulin may play an important central role in the growth and differentiation of this parasite. We have also assessed the calmodulin content of the various life stages by immunoblot analysis. These studies identified a 14-kDa immunoreactive peptide present at equivalent levels in epi-, trypo-, and amastigote stages (extracellular).

Animals↗

Trypanosoma cruzi: mechanisms of intracellular calcium homeostasis.

Regulation of intracellular Ca2+ homeostasis was characterized in epimastigote forms of Trypanosoma cruzi using the fluorescence probe Fura-2. Despite an increase in extracellular Ca2+, [Ca2+]o, from 0 to 2 mM, cytosolic Ca2+, [Ca2+]i, increased only from 85 +/- 9 to 185 +/- 21 nM, indicating the presence of highly efficient mechanisms for maintaining [Ca2+]i. Exposure to monovalent Na+ (monensin)-, K+ (valinomycin, nigericin)-, and divalent Ca2+ (ionomycin)-specific ionophores, uncouplers of mitochondrial respiration (oligomycin), inhibitors of Na+/K(+)-ATPase (ouabain), and Ca(2+)-sensitive ATPase (orthovanadate) in 0 or 1 mM [Ca2+]o resulted in perturbations of [Ca2+]i, the patterns of which suggested both sequestration and extrusion mechanisms. Following equilibration in 1 mM [Ca2+]o, incubation with orthovanadate markedly increased [Ca2+]i, results which are compatible with an active uptake of [Ca2+]i by endoplasmic reticulum. In contrast, equilibration in 0 or 1 mM [Ca2+]o did not influence the relatively smaller increase in [Ca2+]i following incubation with oligomycin, suggesting a minor role for the mitochondrial compartment. In cells previously equilibrated in 1 mM [Ca2+]o, exposure to monensin or ouabain, conditions known to decrease the [Na+]o/[Na+]i gradient, upon which the Na+/Ca2+ exchange pathways are dependent, markedly increased [Ca2+]i. In a complementary manner, decreasing the extracellular Na+ gradient with Li+ increased [Ca2+]i in a dose-dependent manner. Finally, the calcium channel blockers verapamil and isradipine inhibited the uptake of Ca2+ by greater than 50%, whereas diltiazem, nifedipine, and nicardipine were ineffective. The results suggest that epimastigote forms of T. cruzi maintain [Ca2+]i by uptake, sequestration, and extrusion mechanisms, with properties common to eukaryotic organisms.

Animals↗

Intestinal parasites in returned travelers.

Travelers returning from third-world countries may become infected with a variety of intestinal parasites. Although protozoan infections are more frequently seen, intestinal worms are also encountered. If considered in the differential diagnosis, these infections usually are readily diagnosed and treated.

Humans↗

Imported malaria in the Bronx: review of 51 cases recorded from 1986 to 1991.

The cases of 51 patients with malaria seen at the Albert Einstein College of Medicine hospitals from January 1986 to June 1991 are reviewed. Thirty-five patients acquired infection on journeys to their country of origin. Of these 35 patients, 83% of whom had lived in the United States for > or = 2 years, only 17% received antimalarial prophylaxis. Ten of the 51 patients were born and raised in the United States, and 70% received prophylaxis (P < .01). Six of the 51 patients were visitors to the United States from areas endemic for malaria. Overall, 64% of patients acquired malaria in West Africa, south of the Sahara; 20% in Asia; 8% in Ecuador; 6% in Haiti; and 4% in the Middle East. The majority of infections were due to Plasmodium falciparum. Six patients traveled to a zone endemic for malaria while pregnant, and none received prophylaxis. In nine of 13 patients who received prophylaxis, there was inadequate dosing or poor compliance. Individuals born in regions endemic for malaria are at high risk of acquiring malaria on return to their countries of origin and are less aware of the need for malaria prophylaxis than are other travelers.

Adolescent↗

Cytokine gene expression of endothelial cells infected with Trypanosoma cruzi.

Coronary microvascular spasm and platelet hyperreactivity have been implicated in the pathogenesis of Chagas' cardiomyopathy. To clarify further the role of the microvasculature in this disease, alterations in cytokine gene expression due to Trypanosoma cruzi infection of human umbilical vein endothelial cells were examined. Northern blot analysis of total RNA from endothelial cells demonstrated that interleukin (IL)-1 beta, IL-6, and colony-stimulating factor 1 (CSF-1) mRNA expression was absent or minimal in uninfected cells but significantly increased in infected cells. c-sis mRNA levels diminished with increased time of infection. In situ hybridization studies also demonstrated high levels of IL-6 mRNA in individual infected cells. Significant levels of IL-6 and IL-1 beta protein were detected in the supernatants of infected endothelial cells. The serum of an acutely infected individual contained high levels of IL-6 protein, suggesting the potential importance of cytokines secreted by the vascular endothelium in the pathogenesis of Chagas' cardiomyopathy.

Animals↗