Reverse actin sliding triggers strong myosin binding that moves tropomyosin.

Actin/myosin interactions in vertebrate striated muscles are believed to be regulated by the "steric blocking" mechanism whereby the binding of calcium to the troponin complex allows tropomyosin (TM) to change position on actin, acting as a molecular switch that blocks or allows myosin heads to interact with actin. Movement of TM during activation is initiated by interaction of Ca(2+) with troponin, then completed by further displacement by strong binding cross-bridges. We report x-ray evidence that TM in insect flight muscle (IFM) moves in a manner consistent with the steric blocking mechanism. We find that both isometric contraction, at high [Ca(2+)], and stretch activation, at lower [Ca(2+)], develop similarly high x-ray intensities on the IFM fourth actin layer line because of TM movement, coinciding with x-ray signals of strong-binding cross-bridge attachment to helically favored "actin target zones." Vanadate (Vi), a phosphate analog that inhibits active cross-bridge cycling, abolishes all active force in IFM, allowing high [Ca(2+)] to elicit initial TM movement without cross-bridge attachment or other changes from relaxed structure. However, when stretched in high [Ca(2+)], Vi-"paralyzed" fibers produce force substantially above passive response at pCa approximately 9, concurrent with full conversion from resting to active x-ray pattern, including x-ray signals of cross-bridge strong-binding and TM movement. This argues that myosin heads can be recruited as strong-binding "brakes" by backward-sliding, calcium-activated thin filaments, and are as effective in moving TM as actively force-producing cross-bridges. Such recruitment of myosin as brakes may be the major mechanism resisting extension during lengthening contractions.

Anthelmintic efficacy of albendazole against inhibited larvae of Ostertagia ostertagi.

Albendazole was evaluated for its efficacy against inhibited early 4th-stage larvae of Ostertagia ostertagi for the 3rd consecutive year (April 1979) in naturally infected yearling beef cattle. Mean numbers of O ostertagi developmental stages in 10 nontreated controls were: adults, 11,485; developing 4th-stage larvae, 1,370; and early 4th-stage larvae, 51,500. The mean proportion of inhibited early 4th-stage larvae was 76.8%. Small numbers of Haemonchus sp and Trichostrongylus axei were uniformly present in the controls. At a dosage level of 7.5 mg/kg of body weight in a group of 10 cattle, the percentage of reduction in comparison with controls was: O ostertagi adults, 93.5; developing 4th-stage larvae, 53.4; developing 4th-stage larvae, 30.8. Percentage of reduction against Haemonchus sp and T axei adults was 94.5 and 100.0, respectively. At the larger dosage of 10.0 mg/kg in 10 cattle, percentage of reduction was: O ostertagi adults, 94.8; developing 4th-stage larvae, 18.6. Percentage of reduction against Haemonchus sp and T axei adults was 100.0 for both species. Considerable variability of efficacy against inhibited larvae, as well as developing 4th-stage larvae in half of the animals of each treatment group, was responsible for the poor reductions. Clinical signs of toxicosis were not observed after administration of albendazole.

Efficacy of ivermectin against inhibited larvae of Ostertagia ostertagi.

The efficacy of ivermectin against inhibited early 4th-stage larvae of ostertagia ostertagi and other nematodes of the abomasum and intestinal tract was determined in naturally infected yearling beef cattle. The time when large numbers of inhibited larvae were acquired was determined by monthly slaughter of monitor cattle, beginning in January. In April, 12 animals were removed from pasture and maintained free of further helminth exposure until slaughter (21 days). At 9 days after the cattle were removed from pasture, ivermectin was administered to the principals by subcutaneous injection (200 micrograms/kg); the other 6 animals were given subcutaneous injections of the ivermectin vehicle. both groups were klled and necropsied at 12 days after treatment. Mean numbers of O ostertagi in the 6 controls were: adults, 41,906; developing 4th stage, 73,813; and early 4th stage, 334,965. The mean proportion of early 4th-stage larvae was 73.7%. In the 6 principals (treated with ivermectin), the following reductions were observed: O ostertagi adults, 100%; developing 4th stage, 99.8%; and early 4th stage, 99,9%. Small numbers of dead and degenerated O ostertagi of all developmental stages were recovered from abomasal washings before fixation; few viable worms were recovered.

Further studies on the efficacy of fenbendazole against inhibited larvae of Ostertagia ostertagi.

Fenbendazole, at low dosage levels, was evaluated for its efficacy against inhibited early fourth stage larvae of Ostertagia ostertagi in naturally infected, yearling beef cattle in April. The mean number of inhibited larvae in 10 untreated control cattle was 51,500. At a dosage level of 1 mg per kg (pelleted fenbendazole) per day in feed over five days in 10 cattle, the percentage reduction in comparison to controls was: adults, 85.9 per cent; developing stages, 79.0 per cent; inhibited early fourth stage, 47.6 per cent; other abomasal genera (Haemonchus species and Trichostrongylus axei, 100 per cent). At a dosage level of 5 mg per kg (10 per cent liquid suspension) administered as an oral drench to 10 cattle, the percentage reduction in comparison to controls was: adults, 96.9 per cent; developing stages, 82.9 per cent; inhibited early fourth stage, 74.7 per cent; other genera, 98.6 per cent. Efficacy against parasite genera of the intestinal tract (primarily Cooperia species) was in excess of 94 per cent at both dosage levels. Some variability of efficacy was observed, in that large numbers of surviving inhibited O ostertagi larvae were recovered from two or three animals in treated groups.

DDT (dicophane) and postmenopausal breast cancer in Europe: case-control study.

OBJECTIVE: To examine any possible links between exposure to DDE (1,1-dichloro-2,2-bis (p-chlorophenyl)ethylene), the persistent metabolite of the pesticide dicophane (DDT), and breast cancer. DESIGN: Multicentre study of exposure to DDE by measurement of adipose tissue aspirated from the buttocks. Laboratory measurements were conducted in a single laboratory. Additional data on risk factors for breast cancer were obtained by standard questionnaires. SETTING: Centres in Germany, the Netherlands, Northern Ireland, Switzerland, and Spain. SUBJECTS: 265 postmenopausal women with breast cancer and 341 controls matched for age and centre. MAIN OUTCOME MEASURE: Adipose DDE concentrations. RESULTS: Women with breast cancer had adipose DDE concentrations 9.2% lower than control women. No increased risk of breast cancer was found at higher concentrations. The odds ratio of breast cancer, adjusted for age and centre, for the highest versus the lowest fourth of DDE distribution was 0.73 (95% confidence interval 0.44 to 1.21) and decreased to 0.48 (0.25 to 0.95; P for trend = 0.02) after adjustment for body mass index, age at first birth, and current alcohol drinking. Adjustment for other risk factors did not materially affect these estimates. CONCLUSIONS: The lower DDE concentrations observed among the women with breast cancer may be secondary to disease inception. This study does not support the hypothesis that DDE increases risk of breast cancer in postmenopausal women in Europe.

The regulatory domain of the myosin head behaves as a rigid lever.

The regulatory domain of the myosin head is believed to serve as a lever arm that amplifies force generated in the catalytic domain and transmits this strain to the thick filament. The lever arm itself either can be passive or may have a more active role storing some of the energy created by hydrolysis of ATP. A structural correlate which might distinguish between these two possibilities (a passive or an active role) is the stiffness of the domain in question. To this effect we have examined the motion of the proximal (ELC) and distal (RLC) subdomains of the regulatory domain in reconstituted myosin filaments. Each subdomain was labeled with a spin label at a unique cysteine residue, Cys-136 of ELC or Cys-154 of mutant RLC, and its mobility was determined using saturation transfer electron paramagnetic resonance spectroscopy. The mobility of the two domains was similar; the effective correlation time (tau(eff)) for ELC was 17 micros and that for RLC was 22 micros. Additionally, following a 2-fold change of the global dynamics of the myosin head, effected by decreasing the interactions with the filament surface (or the other myosin head), the coupling of the intradomain dynamics remained unchanged. These data suggest that the regulatory domain of the myosin head acts as a single mechanically rigid body, consistent with the regulatory domain serving as a passive lever.

The epidemiology of Ostertagia ostertagi and other gastrointestinal nematodes of cattle in Louisiana.

The epidemiology of Ostertagia ostertagi and other gastrointestinal nematodes in yearling beef cattle was examined in each of 2 successive years. During each year 50 head of newly weaned beef calves were given a single dose of thiabendazole and then placed on experimental pastures. Twenty-four of the animals were designated for monthly slaughter (n = 2) and analysis of worm population characteristics and 25 were designated for blood and faecal collection and for weighing. Parasite-free tracer calves were grazed alongside the yearling cattle each month (n = 2) throughout the 2 years and were also slaughtered for analysis of worm populations. Faecal egg counts, plasma pepsinogen determinations, herbage larval counts, and animal liveweight changes were recorded monthly. Results of this work substantiated previous observations on seasonal changes of populations of the different nematode genera, but greater continuity and definition of patterns was possible in the present work. Ostertagia ostertagi was the predominant nematode present. While minimal numbers of inhibited larvae were observed from autumn into winter, most of the population was adult at this time. Acquisition of inhibition-prone larvae begins in late winter and peak numbers are acquired between March and June. Little transmission of O. ostertagi occurs between June and September. Trichostrongylus axei had a similar seasonal prevalence to O. ostertagi. A major difference in the 2 years was in the pattern of maturation of inhibited larvae. This occurred early, during May 1981, following below average temperatures in May and above average rainfall in May and June. Overt type II disease was not associated with a smaller level of maturation in autumn 1982. In contrast, maturation of large burdens of inhibited larvae did not occur until autumn 1982 following a protracted period of dry and hot weather from spring. Acute type II disease was observed in autumn 1982. Reasons for the differences in maturation pattern, based on worm counts from yearling cattle and tracer calves and association of these with faecal egg counts, herbage larval counts, clinical condition, and liveweight changes are discussed.