Inhibition of C3 with pegcetacoplan results in normalization of hemolysis markers in paroxysmal nocturnal hemoglobinuria.

Paroxysmal nocturnal hemoglobinuria (PNH) is a rare, acquired hematologic disorder characterized by complement-mediated hemolysis. C5 inhibitors (eculizumab/ravulizumab) control intravascular hemolysis but do not prevent residual extravascular hemolysis. The newly approved complement inhibitor, pegcetacoplan, inhibits C3, upstream of C5, and has the potential to improve control of complement-mediated hemolysis. The PADDOCK and PALOMINO clinical trials assessed the safety and efficacy of pegcetacoplan in complement inhibitor-naïve adults (≥ 18 years) diagnosed with PNH. Patients in PADDOCK (phase 1b open-label, pilot trial) received daily subcutaneous pegcetacoplan (cohort 1: 180 mg up to day 28 [n = 3]; cohort 2: 270-360 mg up to day 365 [n = 20]). PALOMINO (phase 2a, open-label trial) used the same dosing protocol as PADDOCK cohort 2 (n = 4). Primary endpoints in both trials were mean change from baseline in hemoglobin, lactate dehydrogenase, haptoglobin, and the number and severity of treatment-emergent adverse events. Mean baseline hemoglobin levels were below the lower limit of normal in both trials (PADDOCK: 8.38 g/dL; PALOMINO: 7.73 g/dL; normal range: 11.90-18.00 g/dL), increased to within normal range by day 85, and were sustained through day 365 (PADDOCK: 12.14 g/dL; PALOMINO: 13.00 g/dL). In PADDOCK, 3 serious adverse events (SAE) led to study drug discontinuation, 1 of which was deemed likely related to pegcetacoplan and 1 SAE, not deemed related to study drug, led to death. No SAE led to discontinuation/death in PALOMINO. Pegcetacoplan was generally well tolerated and improved hematological parameters by controlling hemolysis, while also improving other clinical PNH indicators in both trials. These trials were registered at www.clinicaltrials.gov (NCT02588833 and NCT03593200).

Antimicrobial resistance in New Zealand: the evidence and a call for action.

AMR has been cited as the most significant health issue of the 21st century with potentially serious consequences for the health of global populations, including New Zealand, and its health system. Proactive approaches to combating AMR through better understanding of the causes will inform measures required to reduce potential threats. The Royal Australasian College of Physicians (RACP) identifies three pathogens where increased resistance is of concern and recommends collaborative responses to prevent emerging threats to New Zealand populations. An international best practice AMR programme would include antimicrobial stewardship (AMS) building on evidence, policy, organisational support, multidisciplinary teams and patient experience. The planned Ministry of Health-led collaborative approach to developing a national strategy and programme will provide sector direction. Implementation will require extensive engagement with the health sector and communities to develop joint solutions that prevent further increases in AMR.

Methods for inducing neuronal loss in preweanling rats using intracerebroventricular infusion of kainic acid.

Excitotoxins, such as kainic acid (KA), have been shown to produce neuronal degeneration in the adult rat brain. While preweanling rats have been shown to be relatively resistant to the neurotoxicity of lower doses of KA, the presence of neuronal loss at higher doses (of KA) has only begun to be investigated in such animals. A reliable method of producing neuronal loss in preweanling rats is to administer nmol concentrations of KA via intracerebroventricular (i.c.v.) injections on postnatal day 7 (P7). Using a three-dimensional, non-biased cell counting technique, we have shown that neuronal loss is observed in the CA3 subfield of the hippocampal formation at P45 and P75. Further, immunohistochemical studies of markers for cell death may be useful to examine the types of cellular processes associated with such neuronal loss. Data from our own experiments suggest the activation of immediate-early genes in the neuronal loss produced by KA administration at P7. This developmental animal model of neuronal loss may be useful in studying neurodevelopmental disorders where the onset of symptoms or cognitive deficits is thought to follow an early developmental insult.

Direct current electrical stimulation increases the fusion rate of spinal fusion cages.

STUDY DESIGN: A randomized experimental evaluation of direct current stimulation in a validated animal model with an experimental control group, using blinded radiographic, biomechanical, histologic, and statistical measures. OBJECTIVES: To evaluate the efficacy of the adjunctive use of direct current stimulation on the fusion rate and speed of healing of titanium interbody fusion cages packed with autograft in a sheep lumbar interbody fusion model. SUMMARY OF BACKGROUND DATA: Titanium lumbar interbody spinal fusion cages have been reported to be 90% effective for single-level lumbar interbody fusion. However, fusion rates are reported to be between 70% and 80% in patients with multilevel fusions or with risk factors such as obesity, tobacco use, or metabolic disorders. The authors hypothesized that direct current stimulation would increase the fusion rate of titanium interbody fusion cages packed with autograft in a sheep lumbar interbody fusion model. METHODS: Twenty-two sheep underwent lumbar discectomy and fusion at L4-L5 with an 11- x 20-mm Bagby and Kuslich (BAK) cage packed with autograft. Seven sheep received a BAK cage and no current. Seven sheep had a cage and a 40-microA current applied with a direct current stimulator. Eight sheep had a BAK cage and a 100-microA current applied. All sheep were killed 4 months after surgery. The efficacy of electrical stimulation in promoting interbody fusion was assessed by performing radiographic, biomechanical, and histologic analyses in a blinded fashion. RESULTS: The histologic fusion rate increased as the direct current dose increased from 0 microA to 40 microA to 100 microA (P < 0.009). Histologically, all animals in the 100-microA group had fusions in both the right and left sides of the cage. Direct current stimulation had a significant effect on increasing the stiffness of the treated motion segment in right lateral bending (P < 0.120), left lateral bending (P < 0.017), right axial rotation (P < 0.004), left axial rotation (P < 0.073), extension (P < 0.078), and flexion (P < 0.029) over nonstimulated levels. CONCLUSION: Direct current stimulation increased the histologic and biomechanical fusion rate and the speed of healing of lumbar interbody spinal fusion cages in an ovine model at 4 months.

Adult ontogeny of rat working memory of social interactions.

BACKGROUND: Social memory has features that may make it uniquely appropriate for studying normal aging. We used a cross-section experimental design with an animal model to survey the lifetime adult ontogeny of memory of a brief social interaction. METHODS: Groups of healthy adult male rats representing young adulthood (5 months), middle age (10 months), or old age (19-27 months) were tested weekly over a month in two paradigms. Basic social memory and social interference memory were quantified by differences between investigation times of a juvenile rat during a 5-min interaction (acquisition trial) and a second exposure (recall testing), with interexposure intervals (IEI) ranging from 15 min to 24 h. RESULTS: Although basic social memory of all age groups was similar at the brief or longer IEI, there were memory declines at intermediate IEI delays in older males, especially the oldest groups. Decrements to working memory appeared as early as middle-age when an unfamiliar juvenile was inserted between acquisition and recall testing. Nonetheless, our healthy old animals retained a robust ability to recall identity of a conspecific that a minute by minute comparison suggested involves similar behavioral means of gathering social information. CONCLUSIONS: Normative aging of working social memory in male rats can be characterized as being more fragile, beginning at middle age but without significant further decline until near the end of the life span. Functional impact of these age-dependent changes in social memory, on the other hand, may be minimal for all but the very oldest animals in the social group.

Arterial uptake of biodegradable nanoparticles: effect of surface modifications.

Restenosis is the reobstruction of an artery following interventional procedures such as balloon angioplasty or stenting. Local pharmacotherapeutic approaches using controlled release systems are under investigation to inhibit the regional pathophysiologic process of restenosis. We have been investigating biodegradable nanoparticles (100 +/- 39 nm in diameter, mean +/- sd) for the local intra-arterial drug delivery. The purpose of this study was to investigate nanoparticle surface modifications (see Table 1) to enhance their arterial uptake. The PLGA (polylactic polyglycolic acid copolymer) nanoparticles were formulated by an oil-in-water emulsion solvent evaporation technique using a 2-aminochromone (U-86983, Upjohn and Pharmacia) (U-86) as a model antiproliferative agent. The various formulations of nanoparticles were evaluated for the arterial wall uptake by using an ex-vivo dog femoral artery model. The selected formulations were then tested in vivo in acute dog femoral artery and pig coronary artery models. The nanoparticles surface modified with a cationic compound, didodecyldimethylammonium bromide (DMAB), demonstrated 7-10-fold greater arterial U-86 levels compared to the unmodified nanoparticles in different ex-vivo and in-vivo studies. The mean U-86 levels were 10.7 +/- 1.7 microg/10 mg (dog) and 6.6 +/- 0.6 microg/10 mg (pig) in the artery segments ( approximately 2 cm) which were infused with the nanoparticles. The pig coronary studies further demonstrated that the infusion of nanoparticles with higher U-86 loading reduced the arterial U-86 levels, whereas increasing the nanoparticle concentration in the infusion solutions increased the arterial U-86 levels. The biodistribution studies in pigs following coronary arterial administration of nanoparticles demonstrated disposition of U-86 in the myocardium and distally in the liver and the lung. The mechanism of enhanced arterial uptake of the DMAB surface modified nanoparticles seems to be due to the alteration in the nanoparticle surface charge. The unmodified nanoparticles had a zeta potential of -27.8 +/- 0.5 mV (mean +/- sem, n = 5), whereas the DMAB modified nanoparticles demonstrated a zeta potential of +22.1 +/- 3.2 mV (mean +/- sem, n = 5). The adsorption of DMAB to the nanoparticle surface followed the Freundlich isotherm with binding capacity k = 28.1 microg/mg and affinity constant p = 2. 33. In conclusion, surface modified nanoparticles have potential applications for intra-arterial drug delivery to localize therapeutic agents in the arterial wall to inhibit restenosis.

Three electronic state model of the primary phototransformation of bacteriorhodopsin.

The primary all-trans --> 13-cis photoisomerization of retinal in bacteriorhodopsin has been investigated by means of quantum chemical and combined classical/quantum mechanical simulations employing the density matrix evolution method. Ab initio calculations on an analog of a protonated Schiff base of retinal in vacuo reveal two excited states S1 and S2, the potential surfaces of which intersect along the reaction coordinate through an avoided crossing, and then exhibit a second, weakly avoided, crossing or a conical intersection with the ground state surface. The dynamics governed by the three potential surfaces, scaled to match the in situ level spacings and represented through analytical functions, are described by a combined classical/quantum mechanical simulation. For a choice of nonadiabatic coupling constants close to the quantum chemistry calculation results, the simulations reproduce the observed photoisomerization quantum yield and predict the time needed to pass the avoided crossing region between S1 and S2 states at tau1 = 330 fs and the S1 --> ground state crossing at tau2 = 460 fs after light absorption. The first crossing follows after a 30 degrees torsion on a flat S1 surface, and the second crossing follows after a rapid torsion by a further 60 degrees. tau1 matches the observed fluorescence lifetime of S1. Adjusting the three energy levels to the spectral shift of D85N and D212N mutants of bacteriorhodospin changes the crossing region of S1 and S2 and leads to an increase in tau1 by factors 17 and 10, respectively, in qualitative agreement with the observed increase in fluorescent lifetimes.

Contrast radiography in small bowel obstruction: a prospective, randomized trial.

Small bowel obstruction is a common cause of acute abdominal distress, accounting for up to 20% of emergency admissions to surgical services. Although the majority of obstructions will resolve with conservative therapy alone, there are currently no reliable tests for identifying the patients who will require operation. Barium contrast studies have the potential to rapidly identify patients with complete small bowel obstruction, but many surgeons are hesitant to use them for fear of inducing complications. We report the results of a randomized, prospective trial comparing immediate oral barium contrast studies with plain abdominal X-rays in patients presenting with signs and symptoms of small bowel obstruction. End points included time to resolution of the symptoms or operation, total number of hospital days, and morbidity. Sixty-four patients completed the study; of these, 23 received contrast studies and 41 had plain radiographs only. Six of the contrast group (26%) and 11 of the plain radiograph group (27%) ultimately went to operation. Barium contrast studies had a sensitivity of 100% for diagnosing complete obstruction, whereas the sensitivity of serial plain radiographs was only 82%. Among those going to operation, the time from admission to operation was 8.2 hours in the contrast group and 12.4 hours in the plain radiograph group, but this result did not reach statistical significance (p = 0.25). Total hospital days were similar between the two groups (8 vs. 12 days, p = 0.40). There were no complications resulting from the oral administration of barium. Small bowel contrast studies using barium are safe and may shorten the time to operation in patients presenting with signs and symptoms of small bowel obstruction.

Warm water immersion foot: still a threat to the soldier.

We report a significant incidence of warm water immersion foot (WWIF) in a light infantry battalion during a field exercise. Four hundred soldiers belonging to this battalion were surveyed to determine the prevalence of the WWIF syndrome. One hundred forty-nine soldiers surveyed developed the WWIF syndrome. The affected soldiers had a wide range of disability, ranging from mild discomfort to the inability to ambulate. All soldiers had full recovery within 2 weeks of the injuries. Although the syndrome is self-limited, the loss of these soldiers from combat critically impaired the battalion in its mission. We present this report as a reminder that effective preventive measures should be taken prior to field exercises and deployments.

Photoproducts of bacteriorhodopsin mutants: a molecular dynamics study.

Molecular dynamics simulations of wild-type bacteriorhodopsin (bR) and of its D85N, D85T, D212N, and Y57F mutants have been carried out to investigate possible differences in the photoproducts of these proteins. For each mutant, a series of 50 molecular dynamics simulations of the photoisomerization and subsequent relaxation process were completed. The photoproducts can be classified into four distinct classes: 1) 13-cis retinal, with the retinal N-H+ bond oriented toward Asp-96; 2) 13-cis retinal, with the N-H+ oriented toward Asp-85 and hydrogen-bonded to a water molecule; 3) 13,14-di-cis retinal; 4) all-trans retinal. Simulations of wild-type bR and of its Y57F mutant resulted mainly in class 1 and class 2 products; simulations of D85N, D85T, and D212N mutants resulted almost entirely in class 1 products. The results support the suggestion that only class 2 products initiate a functional pump cycle. The formation of class 1 products for the D85N, D85T, and D212N mutants can explain the reversal of proton pumping under illumination by blue and yellow light.

Clade B-based HIV-1 vaccines elicit cross-clade cytotoxic T lymphocyte reactivities in uninfected volunteers.

A fundamental goal of current strategies to develop an efficacious vaccine for AIDS is the elicitation of broadly reactive cytotoxic T lymphocyte (CTL) reactivities capable of destroying virally infected targets. Recent application of recombinant canarypox ALVAC/HIV-1 vectors as vaccine immunogens in HIV-1,-noninfected volunteers has produced CTL responses in a significant number of vaccinees. Using a newly developed targeting strategy, we examined the capacity of vaccine-induced CTL to lyse autologous targets infected with a diverse group of viral isolates. CTL derived from recipients of a canarypox ALVAC/HIV-1 gp160 (MN) vaccine were found capable of lysing autologous CD4+ lymphoblasts infected with the prototypic LAI strain of HIV-1. When tested against autologous targets infected with primary HIV-1 isolates representing genetically diverse viral clades, CTL from ALVAC/gp160 recipients showed both a broad pattern of cytolysis in which viruses from all clades tested were recognized as well as a highly restricted pattern in which no primary isolates, including clade B, were lysed. Differences in the HLA haplotypes of the volunteers immunized with the envelope vector might be a major determinant of the relative breadth of their CTL response. In contrast to ALVAC/gp160 vaccinees, recipients of the ALVAC/HIV-1 immunogen containing envelope as well as gag and protease genes consistently had CTL reactivities effective against a spectrum of primary isolate-infected targets. These studies demonstrate for the first time that clade B-based canarypox vaccines can elicit broad CTL reactivities capable of recognizing viruses belonging to genetically diverse HIV-1 clades. The results also reinforce the impact of viral core elements in the vaccine as well as the pattern of major histocompatibility complex class I allelic expression by the vaccine recipient in determining the relative breadth of the cellular response.

VMD: visual molecular dynamics.

VMD is a molecular graphics program designed for the display and analysis of molecular assemblies, in particular biopolymers such as proteins and nucleic acids. VMD can simultaneously display any number of structures using a wide variety of rendering styles and coloring methods. Molecules are displayed as one or more "representations," in which each representation embodies a particular rendering method and coloring scheme for a selected subset of atoms. The atoms displayed in each representation are chosen using an extensive atom selection syntax, which includes Boolean operators and regular expressions. VMD provides a complete graphical user interface for program control, as well as a text interface using the Tcl embeddable parser to allow for complex scripts with variable substitution, control loops, and function calls. Full session logging is supported, which produces a VMD command script for later playback. High-resolution raster images of displayed molecules may be produced by generating input scripts for use by a number of photorealistic image-rendering applications. VMD has also been expressly designed with the ability to animate molecular dynamics (MD) simulation trajectories, imported either from files or from a direct connection to a running MD simulation. VMD is the visualization component of MDScope, a set of tools for interactive problem solving in structural biology, which also includes the parallel MD program NAMD, and the MDCOMM software used to connect the visualization and simulation programs. VMD is written in C++, using an object-oriented design; the program, including source code and extensive documentation, is freely available via anonymous ftp and through the World Wide Web.

Aging of the brain-testicular axis: reproductive systems of healthy old male rats with or without endocrine stimulation.

To test the hypothesis that endocrine declines in males are incidental to disease, 24 gonadally intact old (22-24 months) rats were selected on the basis of good general health and assigned to one of three groups. One group of aged males was left untreated for comparison with an untreated control group of young adult males. Results from multiple measures of sociosexual behavior and reproductive physiology indicated that endocrine declines in males are not simply a by-product of increased disease incidence with aging. The untreated old animals showed clear decrements on all 13 measures of hypothalamic-pituitary- testicular (HPT) activity. The other two groups of old males were used to compare responsiveness of the aging HPT axis in healthy males to supplements with a typical exogenous (ExT) androgen regimen (300 micrograms testosterone/kg body wt/SC/daily/6 weeks) or to social stimulation (brief daily exposure to an inaccessible estrous female) for additional episodes of endogenous (EnT) hormone. Neither treatment restored our disease-free old male rats to levels approximating those of untreated young adults. Nonetheless, both treatments activated the aging HPT axis. EnT males showed increases in sociosexual behaviors, growth of androgen-sensitive bulbospongiosus muscle, and elevation of epididymal sperm reserves. ExT males, on the other hand, experienced a more foreboding hypertrophy of the ventral prostate gland. Our conclusion is that endocrine aging in males is ubiquitous and inevitable. Still, aged androgen-sensitive systems of healthy old rats retain notable capacity, particularly, for endogenous activation. Evidence points to functional responses by healthy aged males to the presence of sexually receptive females that, although not quantitatively the same, are qualitatively similar to the responses of young adult males.

Inhibition of thrombus formation by endothelin-1 in canine models of arterial thrombosis.

The effect of endothelin-1 (ET-1) on thrombus formation in vivo was evaluated in two well-established canine models of coronary artery thrombosis. First, the possible antithrombotic effect of ET-1 was examined using the cyclic flow reduction (CFR) model of coronary artery stenosis, vascular endothelial cell and intimal smooth muscle cell injury, and periodic acute platelet thrombus formation. Using a rating system of 0 (no inhibition) to 3 (complete inhibition), ET-1 administration at 0.1, 0.5, and 1.0 microgram/kg, i.v. bolus, produced scores of 1.0 +/- 0.2 (n = 10), 1.8 +/- 0.4 (n = 8), and 2.1 +/- 0.3 (n = 7), respectively. ET-1 injection inhibited ex vivo platelet aggregation induced by ADP and U-46619 by 30-60%. When aspirin was administered at 5 mg/kg prior to ET-1 administration at 0.5 microgramoff, ET-1 produced a CFR rating of 2.7 +/- 0.2 (n = 6). However, higher dose aspirin (30 mg/kg, i.v.) significantly inhibited the antithrombotic effect of ET-1 (0.5 +/- 0.5, n = 4). The antithrombotic effect of ET-1 was also examined using an electrolytic injury model of arterial thrombosis. The time required to produce an occlusive thrombus during the experiments in which ET-1 was administered at 10 and 20 ng.kg-1.min-1 was 77 +/- 15 (p < 0.08) and 105 +/- 16 min (p < 0.05), respectively, compared to 44 +/- 5 min when vehicle was infused. Cardiovascular changes following occlusion were not significantly different between dogs given ET-1 and those given vehicle, suggesting that elevated plasma levels of ET-1 did not exacerbate the adverse effects of coronary occlusion. In addition, plasma ET-1 levels were elevated significantly after occlusion in the dogs given vehicle (from 7.4 to 12.4 pg/ml). Taken together, these date provide further evidence to support the notion that ET-1 release during ischemia may be involved in a protective mechanism that impeded thrombus formation in the stenosed coronary artery.

Protection of ischemic/reperfused canine myocardium by CL18/6, a monoclonal antibody to adhesion molecule ICAM-1.

OBJECTIVE: A blocking monoclonal antibody to intercellular adhesion molecule-1 (ICAM-1), CL18/6, previously has been demonstrated to inhibit neutrophil attachment to isolated vascular endothelium and cardiomyocytes. Due to the well known participation of ICAM-1 in the inflammatory responses associated with myocardial ischemia/reperfusion injury, we investigated if CL18/6 could attenuate myocardial ischemia/reperfusion injury in vivo. METHODS: Saline (3-5 ml, i.v., n = 6), non-blocking control MAb CL18/1D8 or CL18/6 (both 0.5 mg kg-1, i.v., n = 4) were administered prior to coronary occlusion (1 h) and subsequent reperfusion (5 h) produced by inflation of a coronary balloon angioplasty catheter in isoflurane-anesthetized, closed-chest dogs. Heart rate and arterial pressure were measured, and regional myocardial blood flow (rMBF), and myeloperoxidase activity (MPO) to index local neutrophil sequestration, were determined. Myocardial infarct size (IS) was evaluated using the tetrazolium staining technique and expressed as a percent of area at risk (AR). RESULTS: Changes in heart rate and arterial pressure were insignificant throughout the experiment. rMBF (mean +/- s.e.m.) in the ischemic subendocardium for each treatment group was: Saline (0.07 +/- 0.02 ml min-1 g-1); CL18/1D8 (0.04 +/- 0.02); CL18/6 (0.06 +/- 0.02). IS/AR% was: saline (37 +/- 3%); CL18/1D8 (39 +/- 9%); CL18/6 (15 +/- 4%*); * = significantly different from CL18/1D8 and saline, P < 0.05. MPO assayed from AR immediately adjacent to the infarct was significantly reduced below infarct MPO only in the CL18/6 treated group-36%). CONCLUSIONS: The results indicate that CL18/6 antagonism of ICAM-1 provided cardioprotection associated with reduced neutrophil activity in vulnerable myocardium, and suggest that ICAM-1 mediated neutrophil sequestration in endangered cardiac tissue is an important mechanism of myocardial ischemia/reperfusion injury.

Molecular dynamics study of the 13-cis form (bR548) of bacteriorhodopsin and its photocycle.

The structure and the photocycle of bacteriorhodopsin (bR) containing 13-cis,15-syn retinal, so-called bR548, has been studied by means of molecular dynamics simulations performed on the complete protein. The simulated structure of bR548 was obtained through isomerization of in situ retinal around both its C13-C14 and its C15-N bond starting from the simulated structure of bR568 described previously, containing all-trans,15-anti retinal. After a 50-ps equilibration, the resulting structure of bR548 was examined by replacing retinal by analogues with modified beta-ionone rings and comparing with respective observations. The photocycle of bR548 was simulated by inducing a rapid 13-cis,15-anti-->all-trans,15-syn isomerization through a 1-ps application of a potential that destabilizes the 13-cis isomer. The simulation resulted in structures consistent with the J, K, and L intermediates observed in the photocycle of bR548. The results offer an explanation of why an unprotonated retinal Schiff base intermediate, i.e., an M state, is not formed in the bR548 photocycle. The Schiff base nitrogen after photoisomerization of bR548 points to the intracellular rather than to the extracellular site. The simulations suggest also that leakage from the bR548 to the bR568 cycle arises due to an initial 13-cis,15-anti-->all-trans,15-anti photoisomerization.

Molecular dynamics study of bacteriorhodopsin and artificial pigments.

The structure of bacteriorhodopsin based on electron microscopy (EM) studies, as provided in Henderson et al. (1990), is refined using molecular dynamics simulations. The work is based on a previously refined and simulated structure which had added the interhelical loops to the EM model of bR. The present study applies an all-atom description to this structure and constraints to the original Henderson model, albeit with helix D shifted. Sixteen waters are then added to the protein, six in the retinal Schiff base region, four in the retinal-Asp-96 interstitial space, and six near the extracellular side. The root mean square deviation between the resulting structure and the Henderson et al. (1990) model measures only 1.8 A. Further simulations of retinal analogues for substitutions at the 2- and 4-positions of retinal and an analogue without a beta-ionone ring agree well with observed spectra. The resulting structure is characterized in view of bacteriorhodopsin's function; key features are (1) a retinal Schiff base-counterion complex which is formed by a hydrogen bridge network involving six water molecules, Asp-85, Asp-212, Tyr-185, Tyr-57, Arg-82, and Thr-89, and which exhibits Schiff base nitrogen-Asp-85 and -Asp-212 distances of 6 and 4.6 A; (2) retinal assumes a corkscrew twist as one views retinal along its backbone; and (3) a deviation from the usual alpha-helical structure of the cytoplasmic side of helix G.

Induction of neointimal hyperplasia by coronary angioplasty balloon overinflation: comparison of feeder pigs to Yucatan minipigs.

We evaluated the use of simple balloon overinflation to induce neointimal hyperplasia in a porcine model of coronary artery restenosis. By using standard percutaneous transluminal coronary angioplasty techniques, left anterior descending (LAD) and/or left circumflex (LCX) coronary arteries of either juvenile feeder pigs or adult Yucatan minipigs were intentionally overinflated. Four weeks later, resultant neointimal hyperplastic responses (neointima/media area; NI/M) were quantitated morphometrically. At all ballooned sites neointimal hyperplasia occurred only when the internal elastic lamina (IEL) was ruptured; the degree of hyperplasia correlated directly with the injury index, that is, the percentage of IEL circumference that fractured (r = 0.74; n = 25; p < 0.05). Despite similar injury indexes in the LAD bed, there was a trend (p = 0.07; analysis of variance) toward greater NI/M ratios in the Yucatan minipig versus the feeder pig group (1.14 +/- 0.21 vs 0.73 +/- 0.09, n = 7/group). We found no such trend in the LCX bed, where the injury index (25.7% +/- 3.5%) was significantly greater than that of the LAD (18.2% +/- 1.2%, p < 0.05). If variations in balloon-induced vascular injury are accounted for, the technique of balloon overinflation of coronary arteries should prove useful in testing potential antirestenotic agents in either adult or juvenile pigs.

Effects of diets containing gossypol on ovarian histology, function and fertility in prepubertal beef heifers.

Forty-five crossbred beef heifers (weight = 268.3 +/- 5.7 kg) were used to determine the effects of dietary gossypol on ovarian morphology, erythrocyte fragility and fertility. Heifers were randomly assigned to 1 of 3 isonitrogenous dietary treatments. The diet consisted of rice mill feed and milo supplemented with soybean meal (n = 13; control), cottonseed meal (n = 16; low supplementation) which supplied 6.1 g free gossypol animal(-1) day(-1), or whole cottonseed (n = 16; high supplementation) which supplied 13.7 g free gossypol animal(-1) day(-1). The heifers were group-fed each diet for 64 days and were maintained on similar but separate fescue pastures overseeded with wheat. After 64 days, 4 heifers from each diet were confined and fed their respective diets. On Day 10 following estrus, each animal was unilaterally ovariectomized, and the ovary containing the corpus luteum was removed. The remaining ovary was removed 6 to 12 hours after detection of estrus in the next cycle. Erythrocyte fragility increased (P < 0.02) in heifers receiving gossypol compared with that of the controls. Cyclicity in the heifers was 81.3, 68.8 and 38.4% for high, low and control diets, respectively, at the end of the 64-day treatment period. First service conception rate, as determined by palpation per rectum, was similar among treatments (58.3, 33.3, 33.3% for high, low and control groups, respectively). Weight gain increased (P < 0.03) in control heifers compared with that of heifers receiving gossypol. Gross ovarian morphology and histology were similar for all heifers. Although gossypol produced mild toxicosis in heifers, no adverse reproductive effects could be detected from gossypol intake.