Microsomal oxidase activities in relation to age and chlorcyclizine induction in American cockroach,Periplaneta americana,fat body,midgut and hindgut

Female adult American cockroaches, Periplaneta americana L., showed definite age-dependent changes in levels of activity of the microsomal mixed-function oxidases. Cytochrome P-450 levels, EPN-detoxication, and p-nitroanisole O-demethylation activities were very low in young adult insects but increased steadily reaching a natural peak at about 100 days in fat body and at about 90 days in midgut and hindgut. The activities then declined rapidly reaching levels of young insects at about 130 to 140 days of age. NADPH-neotetrazolium-reductase activity was high in young insects, declined later in adult life, and returned to a peak at about 100 days.Injections of chlorcyclizine, a known microsomal enzyme inducer, significantly increased levels of cytochrome P-450, EPN-detoxication, p-nitroanisole O-demethylation, and NADPH-NT-reductase activities in young cockroaches. The drug injections were effective, however, only before the natural activity peak was reached. Beyond this point the injections had no inductive effect indicating that the microsomal oxidases in this insect are uninducible when normal enzyme levels are falling.NADPH-NT-reductase activity in male cockroaches, while being somewhat higher than in females, showed a similar age-dependent curve with the peak occurring at about 120 days.Age-dependent carbaryl resistance in male and female insects tended to follow levels of the microsomal oxidase activities. Fifty to 60-day-old insects, however, tended to be more resistant to the insecticide than microsomal enzyme levels would indicate.RNA levels of normal female insects showed age-dependent curves similar to those of the microsomal enzyme activities, being low in young adults and reaching a peak at about 100 days. Chlorcyclizine injections had little or no effect on total microsomal RNA levels.     

Behavior of 14C oryzalin in soil and plants

Radiochemical studies of field soil treated with ¹⁴C oryzalin (3,5-dinitro-N⁴,N⁴-dipropylsulfanilamide) indicated that the compound was readily degradable. One year after soil treatment with oryzalin, 45% of the original radioactivity had dissipated, 25% was extractable, and 30% was “soil bound”. The extractable fraction contained oryzalin and several degradation products, some of which were isolated and identified. No single degradation product accounted for more than 3% of the applied oryzalin. The “soil-bound” radioactivity was extractable with hot alkali. No significant radioactive residues were detectable in either seed or forage of soybean and wheat plants. No specific metabolites of oryzalin were identified in soybean plants. Trace amounts of radioactivity found in plant tissue appeared to be associated with the various plant constituents.     

Effect of 2,4,5-T and picloram on the regeneration of blackberry (Rubus procerus P.J. Muell) from root segments

The formation of roots and shoots on root segments of Rubus procerus P.J. Muell was prevented by soaking the segments for 24 h in a 10^?4M solution of 2,4,5-T or a 10^?5M solution of picloram. Shoot numbers were significantly increased after treatment with 10^?9M and 10^?10M 2,4,5-T, but picloram did not cause a significant increase in shoot numbers. Measurement of the concentration of 2,4,5-T in the extracambial tissue showed that roots treated with 10^?4M 2,4,5-T contained 5× 10^?8 mmole 2,4,5-T per mg dry weight, and by extrapolation, roots treated with 10^?9M 2,4,5-T contained 2× 10^?10 mmole/mg dry weight. Action du 2,4,5-T et du piclorame sur la régénération de la ronce (Rubus procerus P.J. Muell) è partir de fragments de racines La formation de racines et de tiges è partir de fragments de racines dc Ruhus procerus P.J. Muell a été supprimée par trempage des fragmenls pendant 24 heures dans une solution a 10^?4M et de 2.4,5-T, ou dans une solution 10^?5M de piclorame. Le nombre de pousses s'est accru significativement après traitement avec le 2,4,5.-T è 10^?9M et 10^?10M, mais le piclorame n'a pas provoqué d'accroissemcnt significatif du nombre de pousses. La mesure de la concentration de 2,4,5-T dans le tissu extra-cambial a montré que les racines trailées avec du 2,4,5-T è 10^?4M contenaient 5×10^?8 mmole de 2.4,5-T par mg de poids sec et par extrapolation, quc les racines traitées avec du 2,4,5-T k 10^?9M devaient contenir 2 × 10^?12 mmole/mg de poids sec. Die Wirkiing von 2,4,5-T und Picloram auf den Wuchs der Wurzehegmenten von Bromheeren (Rubus procerus P.J. Muell). Die Bildung von Wurzeln und Sprossen aus Wurzelsegmen-ten von Ruhu.i procerus P.J. Muell wurde durch 24-stündiges Einlegen der Wurzelstücke in 10^?4M 2,4,5-T bzw 10^?5M Picloram verhindert. Die Anzahl neugebiideter Sprosse wurde nach Einlegen in 10^?9M und 10^?10M 2,4,5-T, nicht jedoch durch Picloram, signifikant erhöht. Im extracambialen Gewebe von Wurzeln, die mit 10^?4M 2,4,5-T behand-elt worden waren, wurden 5×10^?8mMol 2,4,5-T je mg Trockengewiclu bestimmt. Durch Extrapolation wurde ermittclt. dass mit 10^?9M 2,4,5-T behandelte Wurzeln 2× 10^?12mMol/mg Trockengewicht cnthielten.     

Mercapturic acid formation in the metabolism of propachlor,CDAA, and fluorodifen in the rat

When [¹⁴C]F₃-fluorodifen (2,4′-dinitro-4-trifluoromethyl diphenylether), carbonyl-[¹⁴C]CDAA (N,N-diallyl-2-chloroacetamide), and carbonyl-¹⁴C-propachlor (2-chloro-N-isopropylacetanilide) were fed to rats, 57 to 86% of the ¹⁴C was excreted via the urine within 48 hr. Although very little radioactivity was excreted in the feces of CDAA-treated rats, 15–22% of the ¹⁴C was excreted in the feces of propachlor- of fluorodifentreated rats and an average of 8% of the ¹⁴C remained in these rats 48 hr after treatment. Oxidation of the ¹⁴C label to [¹⁴C]O₂ was not a major process in the metabolism of these herbicides. The only major radioactive metabolite present in the 24-h urine of fluorodifen-treated rats, 2-nitro-4-trifluoromethylphenyl mercapturic acid, accounted for 41% of the administered dose of ¹⁴C. In the metabolism of CDAA, the corresponding mercapturic acid accounted for 76% of the dose; it was the only major metabolite present in the 24-h urine. In contrast, three major metabolites were detected in the 24-h urine of propachlortreated rats, and the mercapturic acid accounted for only 20% of the dose. The mercapturic acid of each herbicide was identified by mass spectrometry.     

Antifungal mode of action of triforine

Rapidly growing mycelia of Aspergillus fumigatus treated with 10 μg/ml triforine (N,N′-bis-(1-formamido-2,2,2-trichloroethyl)-piperazine) showed little or no inhibition in dry weight increase prior to 2 h. By 2.5–3 h, triforine inhibited dry weight increase by 85%. The effects of triforine on protein, DNA, and RNA syntheses corresponded to the effect on dry weight increase both in time of onset and magnitude. Neither glucose nor acetate oxidation were inhibited by triforine.Ergosterol synthesis was almost completely inhibited by triforine even in the first hour after treatment. Inhibition of ergosterol synthesis was accompanied by an accumulation of the ergosterol precursors 24-methylenedihydrolanosterol, obtusifoliol, and 14α-methyl-Δ^{8, 24 (28)}-ergostadienol. Mycelia treated with 5 μg/ml of triarimol (α-(2,4-dichlorophenyl)-α-phenyl-5-pyrimidinemethanol) also accumulated the same sterols as well as a fourth sterol believed to be Δ^{5, 7}-ergostadienol.Identification of 4,4-dimethyl-Δ^{8, 24 (28)}-ergostadienol in untreated mycelia indicates that the C-14 methyl group is the first methyl group removed in the biosynthesis of ergosterol by A. fumigatus. The lack of detectable quantities of 4,4-dimethyl-Δ^{8, 24 (28)}-ergostadienol in triforine or triarimol-treated mycelia and the accumulation of C-14 methylated sterols in treated mycelia suggests that both fungicides inhibit sterol C-14 demethylation. The accumulation of Δ^{5, 7}-ergostadienol in triarimol-treated mycelia further implies that triarimol also inhibits the introduction of the sterol C-22(23) double bond.Two strains of Cladosporium cucumerinum tolerant to triforine and triarimol were also tolerant to the fungicide S-1358 (N-3-pyridyl-S-n-butyl-S′-p-t-butylbenzyl imidodithiocarbonate).     

Apoplastic redox metabolism: Synergistic phenolic oxidation and a novel oxidative burst

The plant apoplast is an important mediator of communication between the cell cytoplasm and its surroundings. Plant cell suspensions offer a convenient model system to gain insight into apoplastic physiology. Here, we describe a novel phenomenon that took place when two naturally occurring phenolics were added together to either soybean or tobacco cell suspensions. Acetosyringone (AS) and/or hydroxyacetophenone (HAP), phenolics found in the extracellular/apoplast of tobacco cells, were added to soybean or tobacco cell suspensions undergoing an oxidative burst. Individually, AS appeared to be utilized as a typical peroxidase substrate to scavenge hydrogen peroxide, while HAP was utilized at a much lower rate. However, when added together the rate of utilization of both phenolics increased and surprisingly resulted in the production of hydrogen peroxide. We have further characterized this novel phenomenon in suspension cells. This study demonstrates that certain phenolics in plants can cause co-oxidation which, as in animals, could alter the structure and bioactivity of surrounding phenolics.     

In ovo embryotoxicity of α-endosulfan adversely influences liver and brain metabolism and the immune system in chickens

There is a lack of laboratory-based embryonic chicken toxicity studies with the ecologically relevant low dose/s of endosulfan that utilizes a more practical approach such as the chorioallontoic membrane (CAM) injection. In this investigation, 2μg AR grade α-endosulfan/egg (40% of LD₅₀ for embryos) was injected through the CAM in 12-day-old chicken embryos and the activities of glucose-6-phosphatase (G6Pase, EC 3.1.3.9), fructose 1,6-diphosphatase (FDPase, EC 3.1.3.11), adenosine triphosphatase (ATPase, EC 3.6.1.3) and succinic dehydrogenase (SDH, EC 1.3.99.1) and DNA and RNA content in liver and brain tissues and acetyl cholinesterase (AChE, EC 3.1.1.7) in the latter were determined at 24, 48, and 72 h post-exposure. The wet weight of the embryos did not differ between groups. Following endosulfan exposure, except increase in the hepatic ATPase activity (P < 0.01), there was a significant decrease in the following parameters: G6Pase activity in both the liver and brain (P < 0.01), SDH activity in the brain (P < 0.01), brain overall DNA and RNA concentration (P < 0.05), brain AChE activity (P < 0.01). Exposure of 18-day-old embryos to 2-μg endosulfan for 24 h caused decrease (P < 0.01) in the lymphocyte count and IgG content. Histopathology of thymus and bursa of Fabricius revealed a reduction in the population of thymic follicles, smaller thymocytes with the clear vacuoles in cytoplasm and fewer bursocytes accompanied by infiltration of erythrocytes in lymphoid follicles of the endosulfan-treated embryos. It was inferred that in ovo injection of 0.041 μg/g egg weight of α-endosulfan suppress gluconeogenesis (main energy source in embryonic life), nerve transmission, and immunity.     

Comparison of a baiting method and PCR for the detection of Phytophthora fragariae var. rubi in certified raspberry stocks*

In winter 2002/2003, a total of 136 root samples from 57 different raspberry stocks in Scotland were examined for the presence of raspberry root rot caused by the fungus‐like pathogen Phytophthora fragariae var. rubi. All stocks had been planted as propagation material entered at different grades in the Scottish certification scheme or had applied for plant passports. For detection, a modified ‘Duncan bait test’ was compared to a nested PCR method. The two tests identified the same infected stocks: PCR detected 10 positive samples from four different stocks, while the bait test picked up two additional positive samples coming from the same four stocks. The two tests had a similar level of reliability in this examination and a recommendation for one or the other depends mainly on the technical equipment and skills available in the laboratory.     

Cyclooxygenase ‐2 Expression in Mammary Tumors in Dogs and its Correlation to Histologic and Biologic Behavior

Introduction:  Cyclooxygenase‐2 (Cox‐2) is the inducible form and the rate‐limiting enzyme, for conversion of arachidonic acid to prostaglandins. Cox‐2 overexpression, common in carcinomas, is associated with increased growth rate, resistance to apoptosis, angiogenesis, and overall, both local and distant aggressive behavior. Cox‐2 overexpression has been detected in human and canine mammary tumors (MTs). Histopathology of canine MT is not always predictive of biologic behavior, and anecdotally, only 50% of the malignant MTs are expected to metastasize. We hypothesize that Cox‐2 expression correlates with aggressive behavior.
Methods:  This retrospective study evaluated 48 bitches, presented for excision of MT between 2000 and 2003 at FMVZ de Botucatu‐UNESP, Brasil. Follow‐up varied from 18 months to 24 months and included physical examination and thoracic radiographs. Histopathologic examination was performed in all tumors, as well as in metastatic lesions when detected in the follow‐up period. Immunohistochemistry was used to detect expression of Cox‐2 in paraffin blocks (Rabbit polyclonal anti‐PGHS‐2. Oxford Biomedical). 10 adenomas, 10 carcinomas, 10 benign mixed tumors, 10 malignant mixed tumors and 8 cases of primary carcinomas and their metastatic lesions.
Results:  Expression of Cox‐2 varied among groups. Adenomas (32.1%), mixed benign tumors (38%), carcinomas (60.3%), malignant mixed tumors (65.8%), and metastatic carcinomas (81.25%) and their metastatic lesions (84.35%). Statistically significant differences (p < 0.05) were observed between the benign and malignant counterparts and between carcinomas and metastatic carcinomas.
Conclusions:  Cox‐2 expression correlates with both histologic and biologic behavior in mammary carcinomas, and may serve as a predictor of metastatic potential.     

Long‐Term Totally Implantable Venous Access Ports in Dogs and Cats Receiving Chemotherapy

Introduction:  We evaluated the totally implantable subcutaneous vascular access port (VAP) in 16 cancer patients undergoing intermittent chemotherapy for more than 30 months.
Methods:  Ports were surgically placed (The CompanionPort, Norfolk Vet Products, Skokie, Illinois 60076) in the jugular vein of 12 dogs and 4 cats between 1/2002 and 7/2004. Body weight determined polyurethane catheter size (4, 5, 7 fr.). The polysulfone port, surrounded by titanium, was anchored to subcutaneous tissue in the dorsolateral neck and confirmed with C‐arm fluoroscopy. All blood samples were obtained via VAP. Nine anticancer agents, other medications, crystalloids/colloids, and whole blood were administered. Ports were flushed every 4–5 weeks with heparinized saline solution (100 IU/ml). Removed catheters were submitted for bacteriology.
Results:  Seven of 16 animals are still alive. VAP were used for 1.5 to more than 30 months with 4–60 injections/port. Catheter tips were visualized from the left atrium distally into the caudal vena cava. Adverse events included post‐operative subcutaneous bruising and/or hematoma (4/16), difficult aspiration (4/16), catheter malposition (1/16), positional flushing (1/16), and occlusion requiring replacement (1/16). No thrombus formation or extravasation was evident. Bacterial colonization without signs of septicemia was observed in 3/4 catheters.
Conclusions:  VAP are an effective way of achieving long‐term venous access in the dog and cat. Complications are typically minor and infrequent.