Evolutionary and structural analyses uncover a role for solvent interactions in the diversification of cocoonases in butterflies.

Multi-omic approaches promise to supply the power to detect genes underlying disease and fitness-related phenotypes. Optimal use of the resulting profusion of data requires detailed investigation of individual candidate genes, a challenging proposition. Here, we combine transcriptomic and genomic data with molecular modelling of candidate enzymes to characterize the evolutionary history and function of the serine protease cocoonase. Heliconius butterflies possess the unique ability to feed on pollen; recent work has identified cocoonase as a candidate gene in pollen digestion. Cocoonase was first described in moths, where it aids in eclosure from the cocoon and is present as a single copy gene. In heliconiine butterflies it is duplicated and highly expressed in the mouthparts of adults. At least six copies of cocoonase are present in Heliconius melpomene and copy number varies across H. melpomene sub-populations. Most cocoonase genes are under purifying selection, however branch-site analyses suggest cocoonase 3 genes may have evolved under episodic diversifying selection. Molecular modelling of cocoonase proteins and examination of their predicted structures revealed that the active site region of each type has a similar structure to trypsin, with the same predicted substrate specificity across types. Variation among heliconiine cocoonases instead lies in the outward-facing residues involved in solvent interaction. Thus, the neofunctionalization of cocoonase duplicates appears to have resulted from the need for these serine proteases to operate in diverse biochemical environments. We suggest that cocoonase may have played a buffering role in feeding during the diversification of Heliconius across the neotropics by enabling these butterflies to digest protein from a range of biochemical milieux.

Activation of purified cardiac ryanodine receptors by dihydropyridine agonists.

Prior observations have raised the possibility that dihydropyridine (DHP) agonists directly affect the sarcoplasmic reticulum (SR) cardiac Ca(2+) release channel [i.e., ryanodine receptor (RyR)]. In single-channel recordings of purified canine cardiac RyR, both DHP agonists (-)-BAY K 8644 and (+)-SDZ202-791 increased the open probability of the RyR when added to the cytoplasmic face of the channel. Importantly, the DHP antagonists nifedipine and (-)-SDZ202-791 had no competitive blocking effects either alone or after channel activation with agonist. Thus there is a stereospecific effect of SDZ202-791, such that the agonist activates the channel, whereas the antagonist has little effect on channel activity. Further experiments showed that DHP agonists changed RyR activation by suppressing Ca(2+)-induced inactivation of the channel. We concluded that DHP agonists can also influence RyR single-channel activity directly at a unique allosteric site located on the cytoplasmic face of the channel. Similar results were obtained in human purified cardiac RyR. An implication of these data is that RyR activation by DHP agonists is likely to cause a loss of Ca(2+) from the SR and to contribute to the negative inotropic effects of these agents reported by other investigators. Our results support this notion that the negative inotropic effects of DHP agonists result in part from direct alteration in the activity of RyRs.

Giant fundic polyp complicating attenuated familial adenomatous polyposis.

This case details the development of a rapidly growing polypoid mass in the proximal stomach in a patient with known attenuated familial adenomatous polyposis. Surgical resection was required and histology showed hyperplasia with extensive areas of dysplastic adenomatous change. This case illustrates that patients with the attenuated form of familial adenomatous polyposis are at risk for multiple neoplasia distinct from those patients with the classic form of familial adenomatous polyposis.

Modification of cardiac Na(+) current by RWJ 24517 and its enantiomers in guinea pig ventricular myocytes.

We examined the effects of the cardiotonic agent RWJ 24517 (Carsatrin, racemate) and its (S)- and (R)-enantiomers on action potential duration, Na(+) current (I(Na)), and delayed rectifier K(+) current (I(K)) of guinea pig ventricular myocytes. RWJ 24517 (0. 1 and 1 microM) prolongation of action potential duration could not be accounted for by suppression of either the rapid (I(Kr)) or slow (I(Ks),) component of I(K), although RWJ 24517 did reduce I(Kr) at concentrations of 1 microM. A more dramatic effect of RWJ 24517 (0.1-1 microM) and the (S)-enantiomer of RWJ 24517 (0.1-3 microM) was an increase in peak I(Na) and slowing of the rate of I(Na) decay, eliciting a large steady-state current. Neither RWJ 24517 nor the (S)-enantiomer affected the fast time constant for I(Na) decay, but both significantly increased the slow time constant, in addition to increasing the proportion of I(Na) decaying at the slow rate. Both agents elicited a use-dependent decrease of peak I(Na) (3-10 microM), which probably resulted from a slowing of both fast and slow rates of recovery from inactivation. In contrast, the (R)-enantiomer of RWJ 24517 did not induce a steady-state component I(Na) or increase peak I(Na) up to 10 microM, but it decreased peak I(Na) at 30 microM. The (R)-enantiomer displayed little use-dependent reduction of I(Na) during trains of repetitive pulses and had no effect on rates of inactivation or recovery from inactivation. These actions of the racemate and the (S)-stereoisomer to slow inactivation and to prolong both Na(+) influx and action potential duration may contribute to the positive inotropic actions of these agents because the resulting accumulation of intracellular Na(+) would increase intracellular Ca(2+) via Na(+)/Ca(2+) exchange.

Plan and operation of the 1995 National Survey of Family Growth.

OBJECTIVES: This report describes how the 1995 National Survey of Family Growth (NSFG) was designed, planned, and implemented. The NSFG is a national survey of women 15-44 years of age designed to provide national estimates of factors affecting pregnancy and birth rates and the health of women and infants. Planning for the 1995 NSFG began in 1990 at a formal conference with the survey's data users. Suggestions for substantial changes and improvements in the survey were made there and carried out by NSFG staff and the NSFG contractor--the Research Triangle Institute (RTI). METHODS: The survey was converted from paper and pencil interviewing to Computer-Assisted Personal Interviewing (CAPI) to improve the quality, consistency, and timeliness of the data. At the same time, event histories of the respondent's work, education, family background, cohabitation, and sexual partners were added to lend explanatory power to the survey. These changes made the interview and the CAPI program long--average interview length was 103 minutes--and complex, but the CAPI program worked very well. RESULTS: About 260 female interviewers were trained for 7 days in January 1995. These interviewers completed a total of 10,847 interviews with women 15-44 years of age, for a response rate of 79 percent. This report describes how the survey was planned and designed and how the data were collected, edited, and processed for public use. This report may be of interest to NSFG data users and to those planning other computer-assisted surveys.

Characteristics of cocaine block of purified cardiac sarcoplasmic reticulum calcium release channels.

We have examined the effects of cocaine on the SR Ca2+ release channel purified from canine cardiac muscle. Cocaine induced a flicker block of the channel from the cytoplasmic side, which resulted in an apparent reduction in the single-channel current amplitude without a marked reduction in the single-channel open probability. This block was evident only at positive holding potentials. Analysis of the block revealed that cocaine binds to a single site with an effective valence of 0.93 and an apparent dissociation constant at 0 mV (Kd(0)) of 38 mM. The kinetics of cocaine block were analyzed by amplitude distribution analysis and showed that the voltage and concentration dependence lay exclusively in the blocking reaction, whereas the unblocking reaction was independent of both voltage and concentration. Modification of the channel by ryanodine dramatically attenuated the voltage and concentration dependence of the on rates of cocaine block while diminishing the off rates to a lesser extent. In addition, ryanodine modification changed the effective valence of cocaine block to 0.52 and the Kd(0) to 110 mM, suggesting that modification of the channel results in an alteration in the binding site and its affinity for cocaine. These results suggest that cocaine block of the SR Ca2+ release channel is due to the binding at a single site within the channel pore and that modification of the channel by ryanodine leads to profound changes in the kinetics of cocaine block.

Modification of cardiac Na+ channels by anthopleurin-A: effects on gating and kinetics.

We used the whole cell patch clamp technique to investigate the characteristics of modification of cardiac Na+ channel gating by the sea anemone polypeptide toxin anthopleurin-A (AP-A). Guinea pig ventricular myocytes were isolated enzymatically using a retrograde perfusion apparatus. Holding potential was -140 mV and test potentials ranged from -100 to +40 mV (pulse duration 100 or 1000 ms). AP-A (50-100 nM) markedly slowed the rate of decay of Na+ current (INa) and increased peak INa conductance (gNa) by 38 +/- 5.5% (mean +/- SEM, P < 0.001, n = 12) with little change in slope factor (n = 12) or voltage midpoint of the gNa/V relationship after correction for spontaneous shifts. The voltage dependence of steady-state INa availability (h infinity) demonstrated an increase in slope factor from 5.9 +/- 0.8 mV in control to 8.0 +/- 0.7 mV after modification by AP-A (P < 0.01, n = 14) whereas any shift in the voltage midpoint of this relationship could be accounted for by a spontaneous time-dependent shift. AP-A-modified INa showed a use-dependent decrease in peak current amplitude (interpulse interval 500 ms) when pulse duration was 100 ms (-15 +/- 2%, P < 0.01, n = 17) but showed no decline when pulse duration was 100 ms (-3 +/- 1%). This use-dependent effect was probably the result of a decrease in the recovery from inactivation caused by AP-A which had a small effect on the fast time constant of recovery (from 4.1 +/- 0.3 ms in control to 6.0 +/- 1.1 ms after AP-A, P < 0.05) but increased the slow time constant from 66.2 +/- 6.5 ms in control to 188.9 +/- 36.4 ms (P < 0.002, n = 19) after exposure to AP-A. Increasing external divalent cation concentration (either Ca2+ or Mg2+) to 10 mM abolished the effects of AP-A on the rate of INa decay. These results demonstrate that modification of cardiac Na+ channels by AP-A markedly slowed INa inactivation and altered the voltage dependence of activation; these alterations in gating characteristics, in turn, caused an increase in gNa presumably by increasing the number of channels open at peak INa. AP-A slows the rate of recovery of INa from inactivation which is probably the basis for a use-dependent decrease in peak amplitude. Finally, AP-A binding is sensitive to external divalent cation concentrations.(ABSTRACT TRUNCATED AT 400 WORDS)

Lipoid proteinosis: Urbach-Wiethe disease.

An otherwise healthy patient with Urbach-Wiethe disease required surgical removal of two 3rd molar teeth. In this multisystem disorder infiltration of the buccal, pharyngeal and laryngeal mucosa may cause difficulties with tracheal intubation and increase the likelihood of trauma. The anaesthetic implications and management are described.

Inputs to the pontine A5 noradrenergic cell group: a horseradish peroxidase study.

The noradrenergic A5 cell group of the caudal ventrolateral pons has been implicated in regulation of cardiovascular activity. The efferent fibers from this cell group have been established, but the sources of afferents into the region have not. Horseradish peroxidase (HRP) was injected into the A5 region in rabbits, or into regions surrounding A5. The pattern of retrograde labeling indicated that several areas known to have a role in the control of cardiovascular function by the brain send projections to and/or through the A5 region.

Temperature elevations in orthopaedic cutting operations.

Temperature measurements were made in bovine cortical bone using two cutting burst at 20,000 and 100,000 rpm and two reciprocating saw blades at 20,000 strokes per minute. A combination of feed rates and depths of cuts were investigated for the cutting burs. Cortical temperatures decreased as a result of increases in feed rate, but increased as the depth of cut increased. Temperature increase due to an increase in rotational speed was dependent upon the particular bur. The cutting forces were observed to increase with feed rate and depth of cut, but to decrease with increased rotational speed. The effect of saw design and irrigation had a significant effect on the cortical temperature. Without irrigation, the bone temperature exceeded temperatures reported to produce thermal necrosis. Clinical tests during total joint replacement indicated saw temperatures exceeding 200 degrees C when irrigation was not used.

Decayed,Missing,And filled teeth among children, United States.

In December 1965 the Division of Health Examination Statistics successfully concluded a survey of the health of the Nation's children aged 6-11 years. The survey, which began in July 1963, was the second of the Health Examination Survey programs, or "cycles ," which, launched successively, produce statistical information about the health of specific segments of the U.S. population. The conduct and operation of the children's cycle closely followed a blueprint prepared for the preceding adult cycle. Examinations were conducted at 40 randomly selected locations in 25 States by means of mobile examination centers manned by physicians, dentists, psychologists, nurses, and technicians. The target population totaled approximately 24 million children (table III, appendix III). It was defined as all noninstitutional U.S. children aged 6-11 living in the United States (including Alaska and Hawaii) except those living on lands reserved for the use of American Indians. To obtain statistically valid estimates about the health of so many people, a probability sample was designed and selected by a complex, scientific procedure (appendix III). The sample consisted of approximately 7,400 children, or about 185 at each location.

Selected examination findings related to periodontal disease among adults, United States - 1960-1962.

In a previous report the Health Examination Survey estimated the prevalence among U.S. adults of inflammatory disease of the anatomic structures that support teeth. Based on examinations conducted during 1960-62, the estimates underscore not only the high prevalence of periodontal disease but also its wide range of distribution throughout the United States. About 44 million adults aged 18-79 years had gingivitis without obvious pocket formation, the report estimated, and about 23 million had chronic destructive disease with one or more pockets diagnostic of advanced periodontal disease. Distinct patterns of distribution prevailed: more disease was generally present in older people than in younger ones, in men than in women, and in Negro adults than in white. In addition, men and women who were economically and educationally more advantaged usually had less periodontal disease than others. Oral hygiene among sample men and women who had one or more of six specified teeth was also evaluated. As foreseen, both the prevalence and severity of periodontal disease increased as oral hygiene worsened. Demographic differences in the distribution of periodontal disease were either largely or fully leveled when allowance was made for variations in oral hygiene. However, among people with equivalent levels of oral hygiene, older men and women had more periodontal disease than younger men and women. Briefly, both age and oral hygiene emerged as important factors, each of which was related independently of the other, in the prevalence and severity of periodontal disease.

An Assessment of the Occlusion of the Teeth of Children 6-11Years, United States.

In December 1965 the Division of Health Examination Statistics successfully concluded a survey of the health of the Nation's children aged 6-11 years. The survey began in June 1963 and was the second of the Health Examination Survey programs, or "cycles," which, launched successively, produce statistical information about the health of specific segments of the United States population. The conduct and operation of the children's cycle closely followed a blueprint prepared for the preceding adult cycle. Examinations were conducted at 40 randomly selected locations in 25 States by means of mobile examination centers manned by physicians, dentists, psychologists, nurses, and technicians. Before a child was examined, information was obtained from the parent of the child, including demographic and socioeconomic data on the household members as well as medical history, behavioral, and related data on the child to be examined. The target population totaled approximately 23.8 million children (table III, appendix III). It was defined as all noninstitutionalized children aged 6-11 living in the United States (including Alaska and Hawaii) except those living on lands reserved for the use of American Indians. To obtain statistically valid estimates about the health of so many people, a probability sample was designed and selected by a complex scientific procedure (appendix III). The sample consisted of approximately 7,400 children, or about 185 at each location.