Efficacy and longevity of nitenpyram against adult cat fleas (Siphonaptera: Pulicidae).

Nitenpyram (Capstar) is a fast acting, orally administered flea treatment that is absorbed into the blood of the host animal and is readily available for uptake by feeding fleas. We examined the efficacy of a single dose of nitenpyram against adult cat fleas, Ctenocephalides felis (Bouché), over several days. We recorded adult flea mortality and flea egg production on treated and untreated cats. Nitenpyram provided 100% kill of all fleas on the host at the time of treatment and for up to 24 h after treatment. Between 24 and 48 h after treatment, there was a 98.6% reduction in adult flea numbers. From 48 to 72 h, there was a 5% reduction in adult fleas. There was a 97% reduction and 95.2% reduction in the number of flea eggs collected from treated versus untreated animals during the first 48 h and from 48 to 72 h, respectively. In addition, we quantified three distinct behavioral responses of infested adult cats treated with nitenpyram to determine the extent of any immediate, overt behavioral responses in treated animals. A significant increase in scratching, biting, licking, and twitching occurred for 5 h. The biting and licking continued for 7 h after treatment. Administration of nitenpyram provides an effective mechanism to eliminate adult fleas from hosts for up to 48 h after treatment.

Response of dogs treated with ivermectin or milbemycin starting at various intervals after Dirofilaria immitis infection.

The response to heartworm infection before preventative programs were started was investigated in 56 dogs. Dogs were infected with third-stage larvae of Dirofilaria immitis and started on preventative programs (monthly treatment) with ivermectin/pyrantel pamoate (IVM/PP) or milbemycin oxime (MO) 3.5, 4.5, 5.5, or 6.5 months after infection. Each time period comprised a group of six dogs treated with IVM/PP and six treated with MO. Thoracic radiographs were obtained prior to infection, at the start of preventative treatment, and at regular intervals until dogs were necropsied 1 year after the preventative was started. All dogs developed radiographic signs of heartworm disease, and all had heartworm-related arterial changes at necropsy. From Day 210 to 330, interstitial lung disease was less severe in dogs started on MO 3.5 months after infection than in dogs given IVM/PP at the same time. Arterial surfaces were more severe at necropsy in the dogs started on MO at 4.5 months than in the dogs started on IVM/PP at the same time. There was increased caudal lobar arterial and interstitial disease in the dogs treated with IVM/PP compared with dogs treated with MO; this was attributed to the death of young worms within the caudal pulmonary arteries. Dogs started on either preventative at 5.5 and 6.5 months after infection had radiographic changes and necropsy evaluations that were similar to those of untreated controls. This study reinforces the recommendation of the American Heartworm Society that mature dogs be evaluated for infection prior to starting a monthly preventative and that any dog that tests positive by a heartworm antigen test receive treatment with an adulticide prior to starting a heartworm preventative program.

Effects of nitroscanate on adult Taenia pisiformis in dogs with experimentally induced infections.

Twenty-four specific-pathogen-free Beagles were each given 50 cysticerci of Taenia pisiformis that had been harvested from experimentally infected rabbits. Quantitative fecal egg counts and fecal screening for recovery of passed segments were performed on postinoculation days 56 through 70. Twenty-three of 24 dogs fed cysticerci developed patent infections. The 23 dogs with patent infections were assigned to 1 of 2 groups and treated with nitroscanate or a placebo 60 days after inoculation. Egg counts in the treated dogs had markedly decreased by the second day after treatment, and by the sixth day after treatment, segments were not found in the feces of any of the treated animals. The control dogs continued to pass eggs and segments in their feces throughout the 9 days after treatment. The dogs were euthanatized and necropsied 70 days after being inoculated. At necropsy, the mean number of scolices recovered from control dogs was 24.6, the mean number of scolices recovered from treated dogs was 0.25. Worms recovered from the control dogs were intact, gravid cestodes. Efficacy of treatment with nitroscanate at a mean dosage of 56 mg/kg of body weight was 98.9%.

Cyromazine pour-on for the prevention of cutaneous myiasis of sheep.

The results of trials on eight farms to assess the efficacy of two pour-on formulations containing cyromazine for the prevention of cutaneous myiasis of sheep are presented; data from trials on sheep with larval implants and on sheep kept in cages with adult flies are also reported. The incidence of cutaneous myiasis was reduced by between 87 per cent and 100 per cent for eight weeks when a formulation containing 6 per cent w/v cyromazine was used at an application rate of 60 to 85 mg of active ingredient/kg bodyweight. When a formulation containing 10 per cent w/v was used at an application rate of 50 to 100 mg/kg, the incidence of the condition was reduced by between 90 per cent and 100 per cent for eight weeks. Studies of sheep with larval implants, using the formulation containing 10 per cent w/v cyromazine at 50, 100 and 150 mg/kg bodyweight gave variable results with some animals at each dose rate having lost protection by the seventh week. When sheep were treated with the formulation containing 10 per cent w/v cyromazine at 50 or 100 mg/kg and exposed to adult flies in fly-proof cages they were completely protected for nine and eight weeks, respectively.

Luteal progesterone synthesis and high-density lipoprotein. Evidence implicating a pronase-sensitive HDL-binding site in the mechanism by which HDL supports luteal progesterone synthesis.

We have shown previously that corpus luteum cells isolated from the superovulated ovaries of rats treated with 4-amino-pyrazolo[3,4-d]pyrimidine constitute a suitable experimental system by which to investigate the mechanism in which plasma high-density lipoprotein (HDL) plays a role in luteal cellular progesterone synthesis. In the present study, the rate of luteal cellular progesterone synthesis was shown to be stimulated by 125I-labelled HDL up to about 70% of the rate achieved in the presence of native HDL. The concentration of HDL needed for half-maximal stimulation of progesterone synthesis in the presence of lutropin was not significantly different irrespective of whether radioiodinated HDL or unlabelled HDL was used. Experimental conditions for studying the binding of 125I-labelled HDL to isolated luteal cells have been defined and cellular binding affinity and binding capacity have been measured. Exposure of the luteal cells to pronase virtually abolished their capacity to bind 125I-HDL and made them unable to respond to added HDL by increasing their rate of progesterone synthesis in the presence of lutropin. Control experiments showed this effect of pronase on cellular progesterone synthesis not to be due to destruction of cellular lutropin receptors, nor to general cellular damage. This evidence supports the view that luteal cellular binding of HDL is part of the mechanism by which HDL acts in luteal progesterone synthesis. Cellular binding capacity and affinity for 125I-labelled HDL were the same irrespective of whether or not lutropin was present during incubation. Furthermore, the binding capacity and affinity of cells from the ovaries of rats not treated with 4-amino-pyrazolo[3,4-d]pyrimidine were the same as in luteal cells isolated from rats that had been treated.

Stimulation of progesterone synthesis, in corpus luteum cells isolated from 4-amino-3,4-d-pyrazolopyrimidine-treated rats, by cholesterol presented in non-lipoprotein form.

We have previously shown that the administration of 4-amino-3,4-d-pyrazolopyrimidine (4-APP) to rats reduces progesterone synthesis by cells subsequently isolated from their corpora lutea, and that plasma high density lipoprotein (HDL) can restore this progesterone synthesis to normal. In this paper we demonstrate that a dispersion of phospholipid and cholesterol, but not other sterols, can enhance this 4-APP-disabled progesterone synthesis to the same level as can HDL, thus providing the first direct evidence that cholesterol is the component of HDL upon which rat corpus luteum depends for its ability to synthesize progesterone.

Philadelphia-positive metaphases in the marrow after bone marrow transplantation for chronic granulocytic leukemia.

A 28-year-old man with Ph-positive chronic granulocytic leukemia (CGL) was treated by high-dose chemoradiotherapy and transplantation of marrow cells harvested from his HLA-identical brother. One year after bone marrow transplantation (BMT) examination of his marrow showed a minority population of Ph-positive cells; their proportion subsequently fell such that 2 years after transplant analysis of marrow cells showed only cytogenetically normal cells. The patient remains clinically normal with a persisting mild lymphocytosis but without hematological evidence of leukemia. We cannot in this patient distinguish between persisting leukemia that later could no longer be recognized and relapse of leukemia that is now suppressed, perhaps only temporarily. This case emphasizes the need for caution in interpreting chromosomal finding after BMT for CGL.

Activation of adenylate cyclase in bovine corpus-luteum membranes by human choriogonadotropin, guanine nucleotides and NaF.

1. Preincubation of luteal membranes with human choriogonadotropin results in the formation of an activated state of adenylate cyclase which is not reversed by washing and which is limited only by the absence of guanine nucleotides, whereas preincubation with GTP yields only a partially activated adenylate cyclase which requires the presence of both GTP and human choriogonadotropin during assay to demonstrate maximal activity. 2. Preincubation of luteal membranes with GTP and human choriogonadotropin does not lead to a synergistic increase in wash-resistant activity. 3. Luteal membranes that had been preincubated with GTP and hormone exhibited a decreasing rate of cyclic AMP synthesis during the adenylate cyclase assay incubation; addition of GTP during the assay incubation reversed the decrease. 4. Membranes that had been preincubated in the absence of guanine nucleotide and hormone showed a ;burst' phase of cyclic AMP synthesis when GTP was present in the assay incubation and a ;lag' phase with p[NH]ppG (guanosine 5'-[beta,gamma-imido]triphosphate) present in the assay. The presence of human choriogonadotropin with either nucleotide in the assay incubation eliminated the curvatures in plots observed with guanine nucleotides alone. 5. Luteal adenylate cyclase was persistently activated by preincubation with p[NH]ppG alone or in combination with human choriogonadotropin; the activation caused by p[NH]ppG alone was still increasing after 70min of preincubation, whereas that caused by p[NH]ppG in the presence of hormone was essentially complete within 10min of preincubation. 6. Luteal adenylate cyclase that had been partially preactivated by preincubation with p[NH]ppG was slightly increased in activity by the inclusion of further p[NH]ppG in the adenylate cyclase assay incubation, but more so with p[NH]ppG and hormone. Human choriogonadotropin alone caused no further increase in the activity of the partially stimulated preparation unless p[NH]ppG was also added to the assay incubation. 7. GTP decreased the activity of adenylate cyclase in membranes that had been partially preactivated in the presence of p[NH]ppG; the decrease in activity was greater when GTP and hormone were present simultaneously in the assay. 8. The results indicate that stable activation states of adenylate cyclase can be induced by preincubation of luteal membranes in vitro with human choriogonadotropin or p[NH]ppG, and that in the presence of p[NH]ppG the hormone may accelerate events subsequent to guanine nucleotide binding. Stable activation of luteal adenylate cyclase by prior exposure to GTP is not achieved. The involvement of GTPase activity and of hormone-promoted guanine nucleotide exchange in the modulation of luteal adenylate cyclase activity is discussed.

The preparation and purification of isolated rat corpus-luteum cells and their use in studying the relationship between cholesterol biosynthesis and the lutropin-stimulated formation of progesterone.

Isolated luteal cells, prepared from superovulated rat ovaries by digestion with collagenase, were subjected to density-gradient centrifugation on Percoll to give a more highly purified preparation of luteal cells than has been reported previously. The cells formed progesterone when incubated in vitro; lutropin stimulated this steroidogenesis. Progesterone formation was linear for at least 2 h; a minimal lutropin concentration of 1.0 ng/ml was needed for stimulation and concentrations of 3.0 and 100 ng/ml gave half-maximal and maximal responses respectively. The cells were unresponsive towards hormones other than lutropin. Exposure to lutropin raised the cellular cyclic AMP concentration, and dibutyryl cyclic AMP, but not dibutyryl cyclic GMP, was as effective in stimulating steroidogenesis as was lutropin. Aminoglutethimide, an inhibitor of cholesterol side-chain cleavage, completely blocked progesterone formation by the cells, showing cholesterol side-chain cleavage to be an obligatory step in steroidogenesis by these cells. Neither the activity of 3-hydroxy-3-methylglutaryl-CoA reductase nor the incorporation of radioactively labelled acetate or mevalonate into cholesterol by cells incubated in vitro were detectable unless the rats had been treated previously with 4-aminopyrazolo[3,4-d]pyrimidine. In cells from rats so treated, compactin was found to block almost completely the incorporation of radioactively labelled acetate, but not of mevalonate, into cholesterol, indicating that this inhibitor acts in corpus luteum in the same way as it does in other tissues. In cells from rats not treated with 4-aminopyrazolo[3,4-d]pyrimidine compactin had no effect on progesterone formation in vitro, showing cholesterol biosynthesis to be unnecessary for the rapid steroidogenic response by luteal cells to lutropin.

Cyclic AMP levels in blood cells from a short-cycle manic-depressive subject.

Erythrocyte and leucocyte numbers and cyclic AMP content were monitored weekly in a mentally defective short-cycle manic-depressive woman. The erythrocyte count remained relatively constant but cyclic AMP content when manic was significantly higher than when depressed, and the mood and biochemical data appeared to have the same periodicity, with coincident peaks and troughs. Total leucocytes were related to mood, with peaks in the manic phase. Mononuclear cyclic AMP content varied without relation to manic or depressive phases.