Search PubMed⌕ Search

Biomedical subjects

J Alexander

Publications and source records attributed to J Alexander.

At least 433 records · Page 24Linked to original sources

Visceral leishmaniasis: resistance to reinfection in the liver following chemotherapy in the BALB/c mouse.

The ability of BALB/c mice to resist reinfection with Leishmania donovani following chemotherapy was studied. BALB/c mice, infected with L. donovani, were treated on Days 7 and 8 postinfection with free, niosomal, or liposomal sodium stibogluconate. It was found that all three drug treatments caused a dramatic reduction in liver parasite burdens as measured on Days 6 and 29 post-treatment. On Day 6 postdrug treatment infection with L. donovani amastigotes, of mice from infected, drug-treated groups, along with age- and sex-matched uninfected controls, showed that at 23 days later, significantly fewer parasites were recovered from the livers of reinfected animals compared with controls given their first infection. Treatment of mice with sodium stibogluconate 6 days prior to a primary infection significantly reduced the number of parasites recovered 14 days later, especially using the carrier form of the drug. In vivo macrophage activity in the liver, as measured by the uptake of radiolabeled horseradish peroxidase immune complex, was significantly raised following stibogluconate treatment of infected but not uninfected mice. These results suggest that a state of resistance persists in the liver of infected mice following chemotherapy which may in part be due to local macrophage activation but also to an unsuspected persistance of the drug.

Animals↗

Is metallothionein involved in deposition of cadmium in bile?

1. By use of our metallothionein (Mt) antibody in Western blotting and techniques for investigating protein metal-binding capacity, the form and content of Mt in bile, collected from cadmium (Cd) treated rats, were studied. 2. It was found that bile is an excretory pathway for Mt from the liver. 3. However, only immunoreactive proteins of higher molecular weight, probably representing polymerized forms of Mt or Mt bound to other proteins, were detected. 4. No monomeric Mt was found. 5. Excretion of Mt is probably not of importance in biliary metal transport.

Animals↗

Do cadmium and dexamethasone induce different types of metallothioneins?

1. Dexamethasone induced thionein in rat liver cross reacts with antibody made towards Cd-thionein from rat liver. 2. Dex-thionein binds Cd. 3. Compared to Cd-thionein, dex-thionein behaves differently on SDS polyacrylamide gels. 4. On molecular filtration on Sephadex G-75 columns, dex-thionein elutes slightly more towards the low molecular weight region than Cd-thionein. 5. Dex-thionein elutes in one peak with two shoulders after DEAE-Sephadex A-50 ion exchange chromatography, while Cd-thionein elutes in two peaks.

Animals↗

Occurrence of various forms of metallothionein in the rat after a short-term cadmium injection regimen.

1. Results are presented showing that the metal-binding metallothionein (Mt) species induced in rat liver in response to Cd administration consist of dimeric and trimeric forms of Mt. A monomeric form might be the first step in the polymerization process. 2. Two proteins (about 8 and 20 kDa mol. wt) are found in hippocampus, but not in brain cortex. 3. These proteins could not be demonstrated to cross-react with our Mt antibody, but the largest of them has strong Cd-binding capacity. 4. Our Mt antibody cross-reacts with a high metal affinity protein present in both brain cortex and hippocampus of twice the mol.wt (20 kDa) of our purified rat liver Mt standard. 5. The results indicate, however, that these Mt like proteins probably emerge from high molecular or membrane bound forms in the cells. 6. A theory is proposed that the predominant polyacrylamide gel band, matching the monomeric, rat liver Mt standard band, seen for all tissues studied in the present work originate from two sources, namely membrane bound and heavy metal induced monomeric form. 7. It is furthermore suggested that those tissues playing an active role in heavy metal metabolism and in protection against toxicity of such metals contain soluble Mts whose active metal-binding forms are oligomers.

Animals↗

Disseminated strongyloidiasis presenting as purpura.

We report a patient with disseminated strongyloidiasis who was being treated with steroids for cerebral edema caused by brain metastases from urinary bladder carcinoma. He had extensive purpura involving the abdomen, arms, and thighs. A skin biopsy specimen showed numerous larvae of Strongyloides stercoralis. Subsequently, rhabdoid larvae of S. stercoralis were isolated in the stool and the sputum. The patient died 2 days later despite thiabendazole therapy.

Diagnosis, Differential↗

Short-chain alkyl esters of L-dopa as prodrugs for rectal absorption.

The bioavailability of L-dopa following rectal administration of a series of short-chain alkyl esters of L-dopa was determined in rats and dogs. The esters were stable (greater than 360 min) to hydrolysis in physiological buffer. In vitro enzymatic hydrolysis of the esters in plasma was species dependent, with the hydrolytic rate being faster in rat plasma (t 1/2 less than 5 min) than dog plasma (t 1/2 = 68-181 min) or human plasma (t 1/2 = 96-238 min). In vivo hydrolysis in dogs, as indicated by the L-dopa plasma profile following intravenous administration of the esters, was very rapid (high extravascular esterase activity). Significant L-dopa bioavailability was observed in rats following rectal administration of the methyl (46%), ethyl (14%), isopropyl (48%), butyl (100%), and 4-hydroxybutyl (13%) esters of L-dopa (rectal L-dopa absorption, less than 5%). In dogs, significant L-dopa bioavailability was also observed for the methyl (28%), isopropyl (30%), butyl (32%), and 4-hydroxybutyl (34%) esters of L-dopa in the presence of carbidopa. The data indicate that these highly water-soluble (greater than 600 mg/ml) esters of L-dopa are potential candidates for controlled-release rectal delivery systems designed to provide more constant plasma L-dopa levels.

Administration, Rectal↗

Reversal of natural killing susceptibility in target cells expressing transfected class I HLA genes.

A number of studies have suggested that resistance of target cells to natural killing (NK) may be correlated with their level of expression of major histocompatibility complex (MHC) class I antigens. To examine this hypothesis directly, a NK-sensitive class I-deficient human B-cell line was transfected with MHC class I genes. The expression of transfected HLA, but not H-2, class I gene products resulted in loss of susceptibility to human NK-mediated conjugation and cytolysis. Furthermore, this protection did not extend to cytotoxicity mediated by interleukin 2-stimulated human NK effector cells.

B-Lymphocytes↗

Studies on the topical treatment of experimental cutaneous leishmaniasis: the therapeutic effect of methyl benzethonium chloride and the aminoglycosides, gentamicin and paromomycin.

BALB/c mice infected with either Leishmania major or Leishmania mexicana were treated twice a day for 10 days with an ointment containing 15% gentamicin or paromomycin, with or without 12% methylbenzethonium chloride (MBCl). It was found that topical application of either paromomycin or MBCl cured the parasite lesion, and that combined treatment with the two compounds had an additive effect. However, after four days' therapy there was a severe inflammatory response at the treatment site, and in most experiments mice relapsed and renewed lesion growth was observed. It is suggested that a non-specific inflammatory reaction may be an important component of the therapeutic response. In further experiments, L. major infected mice treated with paromomycin and MBCl which had cured but not relapsed 58 days after treatment were challenged with a similar dose of the homologous parasite. Lesions developed 16 days post-infection, and the number of parasites recovered from these lesions was similar to that recovered from lesions in control mice. Therefore no protective immunity had been induced by chemotherapy.

Administration, Topical↗

Disposition and metabolism of the food mutagen 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) in rats.

The disposition and metabolism of a common food mutagen, 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), was studied in rats. Five rats of both sexes were given a single oral dose of 14C-labeled MeIQx (3-4 mg/kg body wt). The male rats excreted 36% of the radioactivity and 15% of the mutagenic activity of the dose given in the urine collected during the first 24 h. In the females the corresponding urine contained 41% of the radioactivity and 12% of the mutagenicity. During the next 48 h only 1-3% of the radioactive dose was excreted in urine. The remaining dose was excreted in the feces except of less than 1% that was retained by the tissues after 72 h. The liver and kidney retained more radioactivity than other organs. In a separate study the metabolites of bile, urine and feces of both sexes were investigated. After a single oral dose of 20 mg 14C-labeled MeIQx/kg body wt, three major non-mutagenic metabolites were identified. These were 2-amino-4(or 5)-(beta-D-glucuronopyranosyloxy)-3,8-dimethylimidazo[4,5-f] quinoxaline, 2-amino-3,8-dimethylimidazo[4,5-f]quinoxalin-4(or 5)-yl sulfate and N-(3,8-dimethylimidazo[4,5-f]quinoxalin-2-yl) sulfamate. Another two metabolites present in bile, urine and feces were 2-(beta-D-glucuronopyranosylamino)-3,8-dimethylimidazo[4,5-f ] quinoxaline and 2-amino-8-hydroxymethyl-3-methylimidazo[4,5-f]quinoxalin-4 (or 5)yl sulfate. All metabolites were essentially non-mutagenic. Most of the mutagenicity still present in bile, urine and feces could be explained by unchanged MeIQx. Unchanged MeIQx was the most abundant form excreted in urine.

Animals↗

Metabolism of the food carcinogen 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline in isolated rat liver cells.

2-Amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) was transformed to at least 10 metabolites in suspensions of hepatocytes isolated from Aroclor 1254 treated rats. Combining biochemical data such as effects on MeIQx metabolism of metabolic modulators and incorporation of radioisotopic sulfur with UV, mass and 1H-NMR spectroscopy, we elucidated the structures of six metabolites. About 40% of the MeIQx was transformed to 2-amino-3,8-dimethylimidazo[4,5-f]quinoxalin-4(or5)-yl sulfate. Other oxygenated metabolites were 2-amino-8-hydroxymethyl-3-methylimidazo[4,5-f]quinoxalin-4(or 5)-yl sulfate and 2-amino-4(or5)-beta-D-glucuronopyranosyloxy-3,8-dimethyli midazo[4,5-f] quinoxaline. Evidence was obtained that a glutathione conjugate was formed. This metabolite, and the other oxygenated metabolites were probably formed in P-450 catalyzed reactions. Two metabolites, 2-beta-D-glucuronopyranosylamino-3,8-dimethylimidazo[4,5-f]q uinoxaline and the N-(3,8-dimethylimidazo[4,5-f]quinoxaline-2-yl) sulfamate, were direct conjugates of MeIQx.

Animals↗

Genotoxicity of the food mutagen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP): formation of 2-hydroxamino-PhIP, a directly acting genotoxic metabolite.

Hepatocytes isolated from Aroclor 1254 (PCB) pretreated rats metabolized 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) to a reactive metabolite that induced DNA damage measured by alkaline elution or as increased unscheduled DNA synthesis. PhIP induced mutations in Salmonella typhimurium TA98 and DNA strand breaks and sister chromatid exchange(s) in Chinese hamster V79 cells co-incubated with PCB-hepatocytes. No, or only minor genotoxic, effects were observed when hepatocytes from non-induced rats were used. The bacterial mutagenicity could be inhibited by alpha-naphthoflavone, indicating a role of P-450 in the activation of PhIP. At least eight different metabolites could be separated on HPLC after PhIP had been incubated with PCB-hepatocytes. All of the directly acting mutagenicity towards S.typhimurium TA98 co-eluted with one of the metabolites. The identity of this metabolite was concluded to be 2-hydroxamino-PhIP based on the following evidence: (i) it reduced ferric ion to ferrous ion as hydroxylamines do, (ii) it had an identical UV spectrum and chromatographic properties as a species formed upon reduction of 2-nitro-PhIP by NADPH P-450 reductase. This product displayed a major peak at m/z 241 during thermospray mass spectrometry in the positive-ion mode as would be expected from 2-hydroxamino-PhIP. 2-Hydroxamino-PhIP was directly genotoxic both to TA98 and V79 cells. The genotoxic activity of the medium after removing the hepatocytes remained stable for several hours. Compared to 2-amino-3,4-dimethylimidazo[4,5-f]quinolone (MeIQ), PhIP caused a much larger increase in DNA damage in V79 cells (with hepatocyte activation), whereas MeIQ was more potent with respect to DNA damage induced in hepatocytes and bacteria.

Animals↗

4-(2-amino-1-methylimidazo[4,5-b]pyrid-6-yl)phenyl sulfate--a major metabolite of the food mutagen 2-amino-1-methyl-6- phenylimidazo[4,5-b]pyridine (PhIP) in the rat.

Isolated rat hepatocytes (4 X 10(6) cells/ml metabolized the food mutagen 2-amino-1-methyl-6-phenylimidazo[4,5-b]-pyridine (PhIP) (100 microM) to at least eight different metabolites in a 4 h duration. The major metabolite formed was 4-(2-amino-1-methylimidazo[4,5-b]pyrid-6-yl)phenyl sulfate (4'-PhIP sulfate). Its rate of formation was increased in hepatocytes from PCB pretreated animals in comparison to hepatocytes from untreated animals. One of the other metabolites was the unconjugated derivative of the sulfate (4'-OH-PhIP). This metabolite was also found after in-vitro incubations of rat liver microsomes from PCB or beta-naphthoflavone pretreated animals. The evidence for the proposed structure of the major metabolite is based on [1H]NMR and UV spectroscopy, incorporation of radiolabelled sulfate and arylsulfatase-lability. The formation of 4'-PhIP sulfate was inhibited by the P-450 inhibitors alpha-naphthoflavone and metyrapone and when incubated in sulfate-free medium added the sulfotransferase inhibitor pentachlorophenol. 4'-PhIP sulfate was also the major metabolite of PhIP in the urine of exposed rat.

Animals↗

Visceral leishmaniasis: drug carrier system characteristics and the ability to clear parasites from the liver, spleen and bone marrow in Leishmania donovani infected BALB/c mice.

The efficacy of various sodium stibogluconate formulations against Leishmania donovani has been investigated using a BALB/c mouse model of visceral leishmaniasis. Only one therapy, multiple dosing with drug loaded sonicated vesicles, liposomes or niosomes, was found to be effective against parasites in the liver, spleen and bone marrow. Other treatments significantly reduced parasite liver burdens but either failed to effect spleen and bone marrow parasites, or were effective but toxic. Prophylactic treatment with sodium stibogluconate preparations, six days before infection, reduced parasite multiplication in the liver (free, niosomal and liposomal drug) and the spleen (sonicated, drug loaded niosomes only), but had no suppressive effect on bone marrow parasite burdens compared with controls. These results indicate that in-vivo sodium stibogluconate persists in some compartments at parasiticidal concentrations and that failure to reach this concentration at some sites of infection such as bone marrow, is the cause of treatment failure and relapse.

1,2-Dipalmitoylphosphatidylcholine↗

Influence of Lsh, H-2, and an H-11-linked gene on visceralization and metastasis associated with Leishmania mexicana infection in mice.

Visceral infection and metastatic lesion development following intravenous or subcutaneous inoculation of Leishmania mexicana amastigotes were examined in different B10 congenic mouse strains carrying alternative alleles at Lsh, H-2, or H-11. The results show that, despite a failure to observe any differences in rates of expansion of primary lesions in mice inoculated subcutaneously, each of these genes could be shown to exert some influence during visceralization and metastatic spread of L. mexicana infection. Of particular interest were (i) the continuous advantage observed throughout 160 to 200 days of infection in Lshr versus Lshs mice, (ii) the association between structural gene polymorphism at H-2 and profound visceral and metastatic spread of the parasite producing disease phenotypes akin to diffuse cutaneous leishmaniasis and post-kala-azar dermal leishmaniasis in humans, and (iii) similar effects observed in mice differing at H-11, the functional basis for which involves modified expression of major histocompatibility complex class II molecules. The results are discussed in relation to the human disease and the possibility that homologs for each of these genes regulate leishmanial infections in humans.

Animals↗

Influence of mean pressure on aortic impedance and reflections in the systemic arterial system.

The present investigation sought to determine the extent to which primary changes in mean arterial pressure (MAP) might influence the calculated aortic impedance. In seven open-chest, anesthetized, autonomically blocked dogs, we measured aortic impedance using white-noise analysis at various levels of MAP achieved by adjusting the height of a left atrial reservoir rather than by pharmacological intervention. Impedance spectra so obtained were parameterized according to best-fit wind-kessel models (Ra, Rc, Ca) and three parameters to characterize wave reflections: the frequency at which the phase of the impedance approached or crossed through 0 degree (phi 0), the frequency of the first minimum of the impedance modulus (fmin), and the amplitude of the first oscillation of the impedance modulus (a 1). The dependency of each of these parameters on MAP was investigated. In addition, we calculated the reflection coefficient spectrum (RCS). Results show that windkessel parameters were not significantly influenced by MAP and that the reflection parameters and the RCS depended on increased levels of MAP in a manner consistent with an increase in pulsewave velocity.

Animals↗

Tumors of the pharyngomaxillary space.

A bulging of the soft palate should alert one to the possibility of a mass in the pharyngomaxillary space. Work-up includes CT scan or MRI and possible needle aspiration. Open biopsy should be avoided. Benign, mixed tumors are most commonly encountered. Surgical excision by external approach is generally advocated.

Adenoma↗