Effects of a new tetrahydroisoquinoline (NC-7197) in experimental cardiogenic shock.

Intravenous administration of 1.0 mg/kg of NC-7197 to open and closed-chest anaesthetized dogs immediately after induction of cardiogenic shock by miliary coronary embolization resulted in significant haemodynamic improvement and prolonged survival. These results suggest that NC-7197 may prove to be valuable in the initial management of myocardial infarction complicated by hypotension and shock.

A rat EEG model for evaluating contrast media neurotoxicity.

The electroencephalographic (EEG) effects of intracisternally administered x-ray contrast media were evaluated in rats as a means of assessing neurotoxicity. Rats were ventilated with a mixture of nitrous oxide and oxygen (70/30) sufficient to maintain light anesthesia/analgesia and neuromuscular blockade was induced to prevent movement artifacts. A femoral artery was catheterized for monitoring arterial blood pressure (BP), heart rate, blood gases, and pH. Four 22-gauge stainless steel needle electrodes were inserted underneath the scalp for recording EEG. Approximately 1 hour after the start of EEG recording, test agents were injected via the cisterna magna and rats were placed in a 20 degrees head-down position. EEG and BP were monitored continuously for up to 160 minutes postinjection. Blood gases and pH were monitored periodically. The effects of meglumine iothalamate (IOT), metrizamide (MET), iogulamide (IOG), and ioversol (IOV) were compared at dose levels from 30 to 240 mgI/kg. Normal saline was injected as a control substance and caused no changes in EEG, blood gases, pH, and BP for up to 160 minutes postinjection. IOT (30 mg I/kg) produced profound EEG effects consistent with epileptogenic activity, followed by slowing and subsequent death in 3 of 4 animals. Metrizamide had minimal EEG effects at 30 mg I/kg but at 60 mg I/kg, and 120 mg I/kg produced moderate to severe EEG changes including epileptiform patterns and death in 33% of animals. IOV caused mild EEG abnormalities in 4 of 12 animals at 120 mg I/kg, mild EEG abnormalities in 6 of 11 animals, and moderate EEG abnormalities in 1 of 11 animals at 240 mg I/kg.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of sodium on the fibrillatory potential of ioversol.

The spontaneous ventricular fibrillation (VF) potential of the nonionic contrast media, ioversol (IOV), with and without the addition of sodium was examined during right coronary artery (RCA) injections into anesthetized closed-chest dogs. Protocols included fixed volume (6 mL) and fixed rate (0.4 and 0.6 mL/sec) injections to compare two or more of the following: IOV, IOV + (0.075-0.9% wt/vol) NaCl, and sodium/meglumine diatrizoate (DIA). In these studies, the incidence of VF for IOV alone was either greater that with IOV + NaCl formulations or, if equivalent, the incidence of other arrhythmias was greater with IOV alone than with the sodium formulations. When DIA was included in the comparisons, the incidence of VF was always greater than IOV with or without sodium. There was a sodium-related concentration prolongation in QT interval that, at 0.9% NaCl, approximated that with DIA, even though the incidence of VF for the sodium formulation was 0/15 vs. 6/12 for DIA. Thus, the addition of sodium to IOV appears to reduce the propensity for sponteneous VF in the canine model.

synthesis and biological testing of nonionic iodinated x-ray contrast media.

A series of nonionic x-ray contrast media containing novel linkages between the triiodobenzene and polyhydroxy moieties have been prepared and screened against established criteria. Of the coupler groups examined (amide including dipeptide, carbamate, ureido, and ester), amide showed the greatest overall utility in terms of safety, solubility, stability, and ease of synthesis. Acute toxicity, solubility and stability data are summarized for each series. In the CNS, certain compounds were well tolerated and some were devoid of any excitatory effects. Intrathecal tissue irritant effects were also unremarkable. Most agents exhibited relatively high osmolalities, raising questions as to the influence of this factor on the safety of myelographic agents. Intravascularly, nonionic media appear also to offer advantages over classical ionic media. However, nonionic media can exert positive inotropic effects and exhibit high viscosity at higher iodine concentrations. Nevertheless, overall results continue to demonstrate the radiographic potential of nonionic contrast media.

A field and laboratory evaluation of a commercial ELISA for the detection of Giardia coproantigens in humans and dogs.

A capture enzyme linked immunosorbent assay (CELISA) was evaluated for its ability to detect Giardia coproantigens in the faeces of humans and dogs in the Perth metropolitan area and Aboriginal communities in Fitzroy Crossing, Western Australia. Using zinc sulphate flotation and light microscopy, Giardia cysts and/or trophozoites were observed in 8 of 57 (14%) human stool samples from Perth and 21 of 55 (38%) stool samples from Fitzroy Crossing, after 2 separate examinations. Analysis of diagnostic sensitivity using the ELISA revealed that coproantigens were detected in all 29 human samples (100%) in which Giardia cysts and/or trophozoites were also present. Coproantigens were detected in one further sample from Perth and in 3 samples from Fitzroy Crossing in which no Giardia cyst or trophozoite was observed. The specificity of the test, as defined using Fitzroy Crossing samples free from Giardia, was 91%. The assay did not cross-react with Giardia-free stool samples containing Hymenolepis nana, Entamoeba coli, E. hartmanni, Chilomastix mesnili or Ancylostoma duodenale. Giardia cysts and/or trophozoites were also observed in 11 of 32 dog faecal samples (34%) in Perth and 11 of 29 dog samples (38%) in Fitzroy Crossing, after one zinc sulphate examination. The sensitivity of the ELISA for dogs was 64% and 55% for Perth and Fitzroy Crossing specimens respectively. The specificity was 95% when Fitzroy Crossing samples were used. Other parasites observed in Giardia-free faecal samples from dogs which did not produce a positive reaction with the kit were Ancylostoma caninum, Sarcocystis sp. and Isospora sp.(ABSTRACT TRUNCATED AT 250 WORDS)

Differences in antigen expression within and between 10 isolates of Giardia duodenalis.

In this study indirect immunofluorescence was performed on both live and fixed trophozoites to determine the level of variability in surface antigen expression between 14 Giardia duodenalis isolates, using a monoclonal antibody raised against the Portland 1 isolate (ATCC No. 30888). Subsets of antigen positive cells were detected in 13 isolates ranging in number from < 1% to 50% of the total population. The differences in antigen expression between 10 uncloned isolates did not correlate with genetic differences determined using isoenzyme analysis. Examination of four clones of the Portland 1 isolate showed that all of the progeny expressed surface antigen at significantly different levels to the parent.

Efficacy of albendazole against Giardia and hookworm in a remote Aboriginal community in the north of Western Australia.

The parasitological, clinical efficacy and tolerability of albendazole in the treatment for both giardiasis and hookworm infection in a remote Aboriginal population was investigated. Albendazole at a dose rate of 400 mg daily for 5 days was highly effective in reducing hookworm egg numbers and both Giardia antigen and cysts. The 36.6% prevalence of Giardia prior to treatment fell to 12% between days 6 and 9, 15% for days 10-17 and rose to 28% between days 18 and 30. Tolerability and clinical efficacy were excellent. The effect of albendazole on hookworm was longer lasting than that on Giardia, reducing percent infection from over 76-2% on days 6-9 and zero by day 18-30 despite conditions highly conducive to rapid re-infection. We conclude that albendazole is highly efficacious against both parasites when used as described but that long term community benefit may require additional education programmes to avoid re-infection with Giardia although treatment strategies would seem appropriate for hookworm.

Multilocus enzyme electrophoresis for identification and typing of Treponema hyodysenteriae and related spirochaetes.

An assessment was made of multilocus enzyme electrophoresis as a means of identifying and typing spirochaetes isolated from pigs. Using five enzyme systems, 36 isolates from Australia, the U.K. and the U.S.A. were divided into 12 electrophoretic types or multilocus genotypes, comprising four major, genetically distinct groups. All 26 isolates of Treponema hyodysenteriae fell into one group, members of which showed relatively little genetic diversity. Ten isolates of non-pathogenic spirochaetes fell into three genetically different groups. Although the technique was capable of typing organisms within the groups, it was not always as discriminatory as DNA-restriction endonuclease analysis. Examination of additional enzyme loci should increase the sensitivity of the method for typing and for overall assessment of genetic relationships between spirochaetes.

Modulation of immunocompetence by cyclosporin A, cyclophosphamide or protein malnutrition potentiates murine cytomegalovirus pneumonitis.

Following intranasal infection with murine cytomegalovirus (MCMV), the levels of viral replication in the lungs of susceptible BALB/c mice were enhanced by treatment with cyclophosphamide (CY), or to a greater extent cyclosporin A (CsA) or the Nu/Nu genotype. Focal inflammation was seen 2-4 days after infection in all groups. This was followed by diffuse interstitial pneumonitis which cleared 12-20 days later in the absence of immunosuppression. Although the initial foci of inflammation were less prominent in infected mice treated with CY or CsA, the most severe interstitial pneumonitis was seen 7 days p.i. in mice given CY, whilst CsA-treatment produced focal and disseminated pneumonitis 7-14 days p.i. and Nu/Nu mice exhibited only the focal response. MCMV-infected mice maintained from weaning on a low protein (4% casein) diet also retained higher titres of virus in their lungs than did normally-fed controls, and displayed more prominent focal pneumonitis.

Ribosomal RNA sequencing reveals differences between the genotypes of Giardia isolates recovered from humans and dogs living in the same locality.

A polymerase chain reaction-based method for genotyping Giardia duodenalis isolates using a polymorphic region near the 5' end of the small subunit ribosomal (SSU) RNA gene is described. Analysis was performed using Giardia cysts purified directly from feces. Isolates were collected from humans and dogs living in isolated Aboriginal communities where Giardia infections are highly endemic. This is the first report of the genetic characterization of Giardia from dogs and humans living in the same locality. Comparison of the SSU-rRNA sequences from 13 human and 9 dog isolates revealed 4 different genetic groups. Groups 1 and 2 contained all of the human isolates, whereas groups 3 and 4 consisted entirely of Giardia samples recovered from dogs. One dog sample contained templates from both groups 2 and 3. These results suggest that zoonotic transmission of Giardia infections between humans and dogs does not occur frequently in these communities. The dog-associated SSU-rRNA sequences have not been reported before, suggesting a new G. duodenalis subgroup. A genetic basis for the differences observed between the groups was supported by sequence analysis of 9 in vitro cultured isolates that were placed into the same genetic groups established by enzyme electrophoresis.

Nomenclature and genetic groupings of Giardia infecting mammals.

Giardia is a ubiquitous and well-known enteric parasite affecting humans and a range of domestic and wild mammals. It is one of the most common parasites of domestic dogs and dairy cattle and a frequently recognized waterborne pathogen. Giardiasis is considered to be a re-emerging infection because of its association with outbreaks of diarrhoea in child-care centres. Although only a single species has been recognized as causing disease in humans and most other mammals, molecular characterization of morphologically identical isolates from humans and numerous other species of mammals has confirmed the heterogeneity of this parasite and provided a basis for a clearer understanding of the taxonomy and zoonotic potential of Giardia.

The prevalence of Giardia and other intestinal parasites in children, dogs and cats from aboriginal communities in the Kimberley.

OBJECTIVE: To determine the prevalence of Giardia duodenalis and other intestinal parasites in children, dogs and cats from Aboriginal communities in the west Kimberley region of Western Australia. DESIGN: A four-year parasitological survey of faecal specimens from humans and faecal and intestinal specimens from dogs and cats. SETTING: Local hospital servicing Aboriginal communities surveyed in this study and the Veterinary School, Murdoch University. POPULATION: Children (under 14 years) and adults, as well as dogs and cats, from five Aboriginal communities. RESULTS: G. duodenalis was the most prevalent parasite in children and adults (32.1% in children, n = 361; 12.5% in adults, n = 24). Human infections with Hymenolepis nana (20.5%) and Entamoeba coli (13.0%) were also common. Ancylostoma duodenale (1.3%), Pentatrichomonas hominis (1.0%), Chilomastix mesnili (0.52%), Entamoeba hartmanni (0.52%), Sarcocystis sp. (0.52%), Trichuris trichiura (0.26%), Enterobius vermicularis (0.26%), Strongyloides stercoralis (0.26%) and Isospora belli (0.26%) were present at low rates. Dogs were most commonly infected with Ancylostoma caninum (51.1%) and G. duodenalis (17.0%). Cats were found to have a high prevalence of Ancylostoma tubaeforme (18.2%), Toxoplasma gondii (18.2%), Isospora felis (15.1%) and Spirometra erinacei (15.1%). CONCLUSIONS: This study has shown that children from Aboriginal communities in the west Kimberley region of Western Australia, particularly in the age group one to five years, are commonly infected with intestinal parasites. The dogs and cats in these communities are also infected. The high prevalence rates of Giardia and other enteric parasites in this survey are indicative of poor living conditions and low levels of hygiene. In addition, the high prevalence of hookworm and Giardia infection in dogs and hookworm and Toxoplasma infection in cats is of potential zoonotic significance for humans in these communities.

The prevalence of hookworm infection, iron deficiency and anaemia in an aboriginal community in north-west Australia.

OBJECTIVE: To determine the prevalence of hookworm infections, iron deficiency and anaemia in an Aboriginal community in the north of Western Australia. DESIGN AND SETTING: A cross-sectional survey conducted in 1992, examining faecal specimens and blood samples from Aboriginals and non-Aboriginals in a remote coastal community in the north of Western Australia. PARTICIPANTS: All those living in the community at the time of the survey. MAIN OUTCOME MEASURES: Parasite status and haematological values for haemoglobin, serum iron, ferritin, transferrin and mean red cell volume. RESULTS: Infections with hookworm were present throughout the Aboriginal population (77%; n = 243), with the highest prevalence in children aged 5-14 years (93%; n = 74). Hookworm was not detected in non-Aboriginals (n = 24). Iron deficiency was common throughout the Aboriginal population, especially in children aged 5-14 years (79%; n = 68) and women aged over 14 years (72%; n = 65). Anaemia was highly prevalent among Aboriginal children aged 5-14 years (84% in hookworm-positive children, 75% in hookworm-negative) and women aged over 14 years (63% in hookworm-positive women, 31% in hookworm-negative). Aboriginals over 14 years of age who had hookworm (n = 82) had significantly lower levels of haemoglobin, serum iron and serum ferritin, a lower mean red cell volume and significantly higher transferrin levels than uninfected Aboriginals (n = 38) and non-Aboriginals (n = 19) in the same age group. Hookworm infections were associated with anaemia (P < 0.01) and iron deficiency (P < 0.01) in people over 14 years of age. The species of hookworm, determined after examining 13 larval cultures and two adult worms, was found to be Ancylostoma duodenale. CONCLUSION: Infections with A. duodenale are endemic in Aboriginals in this community, and are likely to contribute to the high prevalence of iron deficiency and anaemia observed in the Aboriginal population, particularly in children and women.

PCR-based DNA fingerprinting of Giardia duodenalis isolates using the intergenic rDNA spacer.

The potential for the non-coding intergenic rDNA spacer (IGS) to DNA fingerprint Giardia duodenalis isolates was investigated. Conserved PCR primers, specific for the flanking large and small rDNA genes, were used to amplify the IGS from 52 in vitro-cultured Giardia isolates. Four distinct IGS-PCR size groups (1.35-1.6 kb) were observed, which correlated closely with the major genetic assemblages established previously for the same isolates using isoenzyme analysis. IGS-PCR size groups A (1.42 kb) C (1.4 kb) and D (1.35 kb) corresponded to isoenzyme assemblage A, and IGS-PCR group B (1.6 kb) to isoenzyme assemblage B. Amplified products from IGS-PCR size groups A and B, which contained 50/52 isolates, were subsequently digested with 8 different restriction enzymes and their profiles compared. Analysis separated isolates within each IGS-PCR size group into 2 distinct clusters which correlated almost exactly with the same genetic groups established previously using isoenzyme electrophoresis. Within each cluster, both methods exhibited a similar capacity to distinguish between Giardia genotypes although they established different genetic relationships between individual isolates. Much of the variability associated with the IGS was attributed to isolates harbouring multiple IGS-sequence types. Restriction analysis of IGS-PCR products amplified from cloned and parent lines of a human isolate BAH 39, which contains multiple IGS variants, showed that trophozoite populations are homogeneous with respect to the types of IGS-variants they maintain. Furthermore, in vitro culture of the cloned isolate BAH39c9 over a 6-year period also failed to reveal variation in IGS-PCR digestion profiles. These results suggest that IGS-PCR RFLP profiles are inherently stable. IGS-PCR analysis was successfully applied to 11 Giardia cyst samples highlighting the potential for this approach to genotype Giardia isolates without the need for in vitro culture.

Ioglucomide: a new nonionic myelographic agent. Preclinical studies.

Ioglucomide, a new iodinated nonionic contrast medium directed primarily toward myelographic use, was subjected to an extensive toxicological examination in animals. In the majority of studies, ioglucomide was compared directly with metrizamide. In some respects, including freedom from production of arachnoiditis, ioglucomide and metrizamide were comparable. However, acute toxicity after intravenous injection or instillation into cerebrospinal fluid was significantly less for ioglucomide. Also, in contrast to metrizamide, ioglucomide produced no evidence of any type of convulsive activity after subarachnoid administration. The improved safety of ioglucomide could not be related to osmolality; therefore, the importance of osmolality for nonionic myelographic agent safety is questioned.

Biodistribution study of Ethiodized Oil Emulsion 13 for computed tomography of the liver and spleen.

Biodistribution studies were conducted with a new intravenous lipoid contrast material currently undergoing clinical trials in four hospitals. The contrast material selectively opacifies the liver and spleen for computed tomographic examination. The experiments were performed on rats with 125I-labeled ethiodized oil emulsion. The study showed that the liver accumulates nearly 80% of the injected iodine within 15 min of the injection and retains a high concentration over 3 h. The second highest concentration was found in the spleen. More than 99% of the iodine is eliminated from the liver and spleen within 48 h, primarily through the kidneys.

Usefulness of liposomes carrying losefamate for CT opacification of liver and spleen.

Losefamate, a hepatobiliary contrast agent, was encapsulated into liposomes to increase its ability to opacify the liver and spleen on computed tomographic (CT) images. Multilamellar lipid vesicles (lecithin, cholesterol, and stearylamine, in 4:1:1 molar ratio) containing iosefamate in their aqueous phase were prepared. Seven dogs received intravenous injections of 100-300 mg I/kg in one of three forms: encapsulated, unencapsulated, or a mixture of the two in equal parts. Animals that received the opaque vesicles had marked opacification of their livers, bile ducts, gallbladders, spleens (maximum 106 H enhancement), and gastrointestinal tracts. Spleen CT values (an indicator of encapsulated material uptake) were always higher in these dogs than in the animals receiving equivalent amounts of unencapsulated iosefamate alone. At the high-dose level, liver uptake of the encapsulated materials was also greater. Liposome-encapsulated hepatobiliary contrast agents are effective liver and spleen opacifiers for CT imaging in the dog.