Predicting carotid artery disease and plaque instability from cell-derived microparticles.

OBJECTIVES: Cell-derived microparticles (MPs) are small plasma membrane-derived vesicles shed from circulating blood cells and may act as novel biomarkers of vascular disease. We investigated the potential of circulating MPs to predict (a) carotid plaque instability and (b) the presence of advanced carotid disease. METHODS: This pilot study recruited carotid disease patients (aged 69.3 ± 1.2 years [mean ± SD], 69% male, 90% symptomatic) undergoing endarterectomy (n = 42) and age- and sex-matched controls (n = 73). Plaques were classified as stable (n = 25) or unstable (n = 16) post surgery using immunohistochemistry. Blood samples were analysed for MP subsets and molecular biomarkers. Odds ratios (OR) are expressed per standard deviation biomarker increase. RESULTS: Endothelial MP (EMP) subsets, but not any vascular, inflammatory, or proteolytic molecular biomarker, were higher (p < .05) in the unstable than the stable plaque patients. The area under the receiver operator characteristic curve for CD31(+)41(-) EMP in discriminating an unstable plaque was 0.73 (0.56-0.90, p < .05). CD31(+)41(-) EMP predicted plaque instability (OR = 2.19, 1.08-4.46, p < .05) and remained significant in a multivariable model that included transient ischaemic attack symptom status. Annexin V(+) MP, platelet MP (PMP) subsets, and C-reactive protein were higher (p < .05) in cases than controls. Annexin V(+) MP (OR = 3.15, 1.49-6.68), soluble vascular cell adhesion molecule-1 (OR = 1.64, 1.03-2.59), and previous smoking history (OR = 3.82, 1.38-10.60) independently (p < .05) predicted the presence of carotid disease in a multivariable model. CONCLUSIONS: EMP may have utility in predicting plaque instability in carotid patients and annexin V(+) MPs may predict the presence of advanced carotid disease in aging populations, independent of established biomarkers.

The APOL1 genotype of African American kidney transplant recipients does not impact 5-year allograft survival.

Apolipoprotein L-1 (APOL1) gene variants are associated with end-stage renal disease in African Americans (AAs). Here we investigate the impact of recipient APOL1 gene distributions on kidney allograft outcomes. We conducted a retrospective analysis of 119 AA kidney transplant recipients, and found that 58 (48.7%) carried two APOL1 kidney disease risk variants. Contrary to the association seen in native kidney disease, there is no difference in allograft survival at 5-year posttransplant for recipients with high-risk APOL1 genotypes. Thus, we were able to conclude that APOL1 genotypes do not increase risk of allograft loss after kidney transplantations, and carrying 2 APOL1 risk alleles should not be an impediment to transplantation.

A novel SNP in COMT is associated with alcohol dependence but not opiate or nicotine dependence: a case control study.

BACKGROUND: It is well established that COMT is a strong candidate gene for substance use disorder and schizophrenia. Recently we identified two SNPs in COMT (rs4680 and rs165774) that are associated with schizophrenia in an Australian cohort. Individuals with schizophrenia were more than twice as likely to carry the GG genotype compared to the AA genotype for both the rs165774 and rs4680 SNPs. Association of both rs4680 and rs165774 with substance dependence, a common comorbidity of schizophrenia has not been investigated. METHODS: To determine whether COMT is important in substance dependence, rs165774 and rs4680 were genotyped and haplotyped in patients with nicotine, alcohol and opiate dependence. RESULTS: The rs165774 SNP was associated with alcohol dependence. However, it was not associated with nicotine or opiate dependence. Individuals with alcohol dependence were more than twice as likely to carry the GG or AG genotypes compared to the AA genotype, indicating a dominant mode of inheritance. The rs4680 SNP showed a weak association with alcohol dependence at the allele level that did not reach significance at the genotype level but it was not associated with nicotine or opiate dependence. Analysis of rs165774/rs4680 haplotypes also revealed association with alcohol dependence with the G/G haplotype being almost 1.5 times more common in alcohol-dependent cases. CONCLUSIONS: Our study provides further support for the importance of the COMT in alcohol dependence in addition to schizophrenia. It is possible that the rs165774 SNP, in combination with rs4680, results in a common molecular variant of COMT that contributes to schizophrenia and alcohol dependence susceptibility. This is potentially important for future studies of comorbidity. As our participant numbers are limited our observations should be viewed with caution until they are independently replicated.

Interrelated striated elements in vestibular hair cells of the rat.

Unusual fixation procedures revealed a series of interrelated striated organelles in type I and type II vestibular hair cells of the rat; these organelles seemed to be less well developed in cochlear hair cells. The findings suggest that contractile elements may play a role in sensory transduction in the inner ear, particularly in the vestibular system.

Development of Squamous Cell Carcinoma After Pulmonary Aspergillosis in the Native Lung of a Lung Transplant Recipient: A Case Report.

Lung transplant recipients have a significant incidence of posttransplant lung nodules. Such nodules can occur from various etiologies, both in the lung allograft or in the native lung. They most commonly originate from infections, such as Pseudomonas or Aspergillus species, or from posttransplant lymphoproliferative disorder. Lung cancer is challenging to diagnose in a native lung, especially with an underlying fibrotic disease. We present a case of a 75-year-old woman who presented with classic clinical features of pulmonary aspergillosis in the native right lung with idiopathic pulmonary fibrosis 5 years after left-sided single-lung transplant. She required a right lower lobectomy and antifungal treatment with isavuconazonium sulfate and inhaled amphotericin. A persistent right upper lobe lung nodule was noted during surveillance imaging and was initially presumed to be recurrent Aspergillus infection; however, growth of the nodule and change in its characteristics prompted additional examination. A navigational bronchoscopic biopsy was positive for squamous cell carcinoma. Her options for stage IIIA squamous cell carcinoma were limited to chemotherapy with paclitaxel and carboplatin plus radiation. Although initial surveillance scans showed adequate tumor response, metastatic squamous cell carcinoma was found in the liver 6 months later. She was eventually transitioned to palliative care. This case highlights the importance of a high index of suspicion for examination of nodules in the native lung of lung transplant recipients, even in cases of a known diagnosis, owing to the high morbidity and mortality associated with primary lung cancer in this population.

Selection with gene-cytoplasm interactions. I. Maintenance of cytoplasm polymorphisms.

General conditions for the protectedness of gene-cytoplasm polymorphisms are considered for a biallelic model with two cytoplasm types and under the assumption that nuclear polymorphisms cannot be maintained in the presence of only one cytoplasm type. Analytical results involving male fertilities, female fertilities, viabilities and selfing rates are obtained, and numerical results show spiral and cyclic behavior of population trajectories. It is shown that a maternally inherited cytoplasmic polymorphism cannot be maintained in the absence of a nuclear polymorphism, and that a gene-cytoplasm polymorphism can only be maintained if the population shows sexual asymmetry, i.e. , if the ratio of male to female fertility varies among genotypes. Thus, the classical viability selection model does not allow gene-cytoplasm polymorphisms.

Selection with Gene-Cytoplasm Interactions. II. Maintenance of Gynodioecy.

Gynodioecy is apparently frequently inherited through gene-cytoplasm interactions. General conditions for the protectedness of gene-cytoplasm polymorphisms for a biallelic model with two cytoplasm types were obtained previously, and these are applied to seven special cases of gene-cytoplasm interactions controlling gynodioecy and involving dominance. It is assumed that nuclear polymorphisms cannot be maintained in one cytoplasm type only. It is held that pure cytoplasmic inheritance of gynodioecy without nuclear interactions is unlikely, and it is shown that gynodioecy with gene-cytoplasm interactions is easier to establish than purely nuclear gynodioecy, for monogenic biallelic dominant or recessive inheritance. For three special cases, a resource-allocation model with simple assumptions always leads to conditions for protectedness of gynodioecy.

Selection in plant populations of effectively infinite size. V. Biallelic models of trioecy.

A one-locus two-allele model of trioecy (presence of hermaphrodites, males and females in one population) is considered, in order to study the conditions for the persistence of this system. All possible assignments of the three sex types to the three genotypes are considered. This leads to three different modes of inheritance of trioecy, namely (a) females heterozygous, (b) males heterozygous and (c) hermaphrodites heterozygous, where in each mode each of the remaining two sex types is homozygous for one of the alleles. For mode (c) trioecy is always persistent, and the dependence of the sex ratio (for the three sex types) on the ovule and pollen fertilities and on the hermaphrodite selfing rate is specified. For the other two modes, (a) and (b), trioecy is not protected, i.e., it may not persist for any fertilities, viabilities or selfing rates. Thus, in this situation it is important to study the conditions under which the "marginal" systems of sexuality of trioecy, i.e., hermaphroditism, dioecy and gynodioecy in mode (a), and hermaphroditism, dioecy and androdioecy in mode (b), may become established. The results show that each marginal system may evolve from each other via trioecy. The evolution of dioecy is easier in mode (a) than in (b), so that female heterogamety would be expected to occur more often than male heterogamety in the present model. Under some conditions the breeding system obtained in equilibrium populations may depend on the initial genotype frequencies.-The necessity of considering modes of inheritance for sexual polymorphisms is demonstrated by comparing our results with those obtained from an evolutionary stable strategy (ESS) analysis of a purely phenotypic model.

Morphological evidence for local microcircuits in rat vestibular maculae.

Previous studies suggested that intramacular, unmyelinated segments of vestibular afferent nerve fibers and their large afferent endings (calyces) on type I hair cells branch. Many of the branches (processes) contain vesicles and are presynaptic to type II hair cells, other processes, intramacular nerve fibers, and calyces. This study used serial section transmission electron microscopy and three-dimensional reconstruction methods to document the origins and distributions of presynaptic processes of afferents in the medial part of the adult rat utricular macula. The ultrastructural research focused on presynaptic processes whose origin and termination could be observed in a single micrograph. Results showed that calyces had 1) vesiculated, spine-like processes that invaginated type I cells and 2) other, elongate processes that ended on type II cells pre- as well as postsynaptically. Intramacular, unmyelinated segments of afferent nerve fibers gave origin to branches that were presynaptic to type II cells, calyces, calyceal processes, and other nerve fibers in the macula. Synapses with type II cells occurred opposite subsynaptic cisternae (C synapses); all other synapses were asymmetric. Vesicles were pleomorphic but were differentially distributed according to process origin. Small, clear-centered vesicles, approximately 40-60 nm in diameter, predominated in processes originating from afferent nerve fibers and basal parts of calyces. Larger vesicles approximately 70-120 nm in diameter having approximately 40-80 nm electron-opaque cores were dominant in processes originating from the necks of calyces. Results are interpreted to indicate the existence of a complex system of intrinsic feedforward (postsynaptic)-feedback (presynaptic) connections in a network of direct and local microcircuits. The morphological findings support the concept that maculae dynamically preprocess linear acceleratory information before its transmission to the central nervous system.

Outcrossing and Sex Function in Hermaphrodites: A Resource-Allocation Model.

Genetic estimates of outcrossing rates in seed plants usually measure the ovule or seed outcrossing rate only. However, there is good evidence for population variation in pollen fertility, which may result in sharply different ovule and pollen outcrossing rates. Therefore, a new combined ovule and pollen outcrossing rate, which depends upon the concept of successful gametes, is proposed. This rate estimates the number of ovules and pollen grains which take part in crossing, as a proportion of all successful ovules and pollen grains (fitness value). Fertility variation also causes unequal male/female functioning of hermaphrodites, and the degree of female function (functional sex) is defined as the number of successful ovules as a proportion of all successful gametes. This is a number between 0 (for males) and 1 (for females), and seldom equals 0.5 for non-fully-selfed hermaphrodites. All of these quantities are frequency dependent, since a common component (the number of successful cross-fertilizing pollen grains) is frequency dependent because of the variation in ovule and pollen fertility. These quantities are studied by means of a one-locus two-allele model of male/female resource allocation, with dominance, in which all genotypes have the same reproductive resources. For 0 < σ < 1, in which σ is the ovule selfing rate, the phenotype with the greater pollen fertility has the greater combined outcrossing rate. For polymorphic populations and 0 < σ < 1, all genotypic combined outcrossing rates are increasing functions of the frequency of the more ovule-fertile phenotype. For σ < 1, functional sex is an increasing and fitness a decreasing function of the frequency of the more pollen-fertile phenotype. Optimal resource allocation for a genotype is defined as that which results in the fixation of that genotype. The genotypic proportion of resources devoted to seeds (Rij) is optimally ½ for σ = 0, and increases with σ. Optimal combined outcrossing and functional sex are similarly defined. Population dynamics can be described in terms of resource allocation as follows: If one genotypic R value is less than and the other greater than ½(1 + σ), then $$\hat{\bar{R}}$$ , the mean seed resource allocation in equilibrium populations, equals ½(1 + σ), whereas if both R's are less than or greater than ½(1 + σ), $$\hat{\bar{R}}$$ equals the genotypic value nearest to ½(1 + σ). Unlike the classical selection model, there may be a polymorphism despite dominance, and mean population fitness may decrease with selection, since polymorphic equilibrium mean fitness is an increasing function of σ. The mean combined outcrossing rate and functional sex equal the ovule outcrossing rate 1 - σ and ½, respectively, in equilibrium populations.

Sites of recombinant adeno-associated virus integration.

Adeno-associated virus (AAV), a defective parvovirus, is considered a promising vector for the delivery of therapeutic genes to cells. Both wild-type and recombinant AAV display a wide tropism and integrate into the host genome, in the absence of helper virus, establishing a latent infection. A unique characteristic of wild-type AAV and a potential advantage for use as a delivery system for gene therapy is the site-specific integration of wild-type virus within a small region of chromosome 19, 19q13.3-qter (AAVS1), in up to 85% of cell lines infected with the virus. Although recombinant AAVs, containing only the inverted terminal repeats of wild-type virus, can integrate efficiently into the host genome, specificity for the AAVS1 site appears to be lost. To address this question, the integration characteristics of two recombinant AAVs lacking the rep and cap genes in HeLa cells were examined. Analysis of Southern blots indicated that none of twenty-six cell clones generated after infection with either one of the recombinant AAVs demonstrated integration within the AAVS1 locus on chromo-some 19. Analysis of five of the cell lines by fluorescent chromosome in situ hybridization confirmed the loss of chromosome 19 specificity. Each integration site mapped near a known fragile site and/or location of a proto-oncogene or growth regulatory gene. Retention of site-specific integration of wild-type AAV will require the inclusion of additional AAV-specific sequences within the recombinant vectors.

Changes in distribution of lung perfusion and ventilation at rest and during maximal exercise.

A new method for evaluation of changes in the distribution of pulmonary perfusion and ventilation during exercise was applied to normal male volunteers. Ventilation and perfusion scans were done with the subjects seated on a bicycle ergometer. The resting studies utilized krypton 81 (81mKr) for ventilation and technetium 99m (99mTc) macroaggregate albumin intravenously for perfusion. Exercise studies were done when 80 percent of maximum predicted heart rate was maintained for five minutes and utilized 81mKr for ventilation and a tenfold dose of 99mTc for perfusion. Higher dose of 99mTc would minimize the effect of radioactivity left over from the resting study. This method allowed us to assess changes in ventilation and perfusion in normal subjects induced by exercise, but may also be applicable in a variety of cardiopulmonary conditions that affect pulmonary ventilation and perfusion or both.

Evidence of a sensory processing unit in the mammalian macula.

We cut serial sections through the medial part of the rat vestibular macula for transmission electron microscopic (TEM) examination, computer-assisted 3-D reconstruction, and compartmental modeling. The ultrastructural research showed that many primary vestibular neurons have an unmyelinated segment, often branched, that extends between the heminode (putative site of the spike initiation zone) and the expanded terminal(s) (calyx, calyces). These segments, termed the neuron branches, and the calyces frequently have spine-like processes of various dimensions with bouton endings that morphologically are afferent, efferent, or reciprocal to other macular neural elements. The major questions posed by this study were whether small details of morphology, such as the size and location of neuronal processes or synapses, could influence the output of a vestibular afferent, and whether a knowledge of morphological details could guide the selection of values for simulation parameters. The conclusions from our simulations are (1) values of 5.0 k omega cm2 for membrane resistivity and 1.0 nS for synaptic conductance yield simulations that best match published physiological results; (2) process morphology has little effect on orthodromic spread of depolarization from the head (bouton) to the spike initiation zone (SIZ); (3) process morphology has no effect on antidromic spread of depolarization to the process head; (4) synapses do not sum linearly; (5) synapses are electrically close to the SIZ; and (6) all whole-cell simulations should be run with an active SIZ.

Computer-assisted three-dimensional reconstruction and simulations of vestibular macular neural connectivities.

The macular neuroepithelium is morphologically organized as a weighted neural network for parallel distributed processing of information. The network is continuous across the striola, where some type II hair cells synapse with calyces containing type I cells with tufts of opposite directional polarities. Whether other hair cell to calyx appositions that lack synapses interact because of intercellular potassium accumulation remains an open question. A functionally important inference of macular organization is that just as arrays of hair cells communicate an entire piece of information to a nerve fiber, so do macular subarrays of nerve fibers (not single units) carry the whole coded message to the brain stem. Moreover, the size of the network subarray can expand or become more limited depending upon the strength and/or duration of the input. It is the functioning of the network and its subarrays that must be understood if we are to learn how maculas carry out their work and adapt to new environments. Simulations of functioning maculas, or subparts, based on precise morphology and known physiology are useful tools to gain insights into macular information processing. The current simulations of afferent collateral electrical activity are a prelude to development of a 3-D model. The simulations demonstrate a relationship between geometry and function, with the diameter of the stem apparently being a major determinant of electrical activity transmitted to the base in the case of collaterals with short stems. Thus, while changes in synaptic number and/or size may be an important adaptive mechanism in an altered g environment, changes in diameter of the stem is another means of altering outflow. Research on the effects of microgravity should be extremely useful in examining the validity of this and other concepts of neural adaptation, since maculas are biological linear accelerometers ideally suited to the task. Maculas are also extremely interesting to study in detail because of the richness of connectivities and submicroscopic organization they present. Many of their features are common with more complex parts of the brain. It seems possible that knowledge of the three-dimensional geometric relationships operative in a functioning macula will contribute much to the understanding of the dynamics underlying more complex behavior. Computerized approaches greatly facilitate this task and provide an objective method of analysis. It is likely that, in the end, simple rules will be found to govern optimal neural architectural organization, even at higher cognitive levels. The architecture only appears complex because we do not yet grasp its meaning.(ABSTRACT TRUNCATED AT 400 WORDS)

Distribution and extent of schistosomiasis in female pelvic organs, with special reference to the genital tract, as determined at autopsy.

PIP: Tissue samples were taken in a series of 64 consecutive autopsies of African women aged 16-70 years. These were block-dissected and studied to determine the frequency of schistosome eggs in tissues of the uterus, and its adnexa, and in the vagina, as well as to determine the numbers of eggs in these tissues. In addition, by histological examination, it was hoped that the frequency and degree of inflammation accompanying the eggs' presence could be learned. 37 of 64 autopsy specimens showed evidence of schistosomiasis. In terms of the mean number of eggs per gram of tissue, the heaviest deposition of eggs was in the bladder. The cervix, uteri, fallopian tubes, and ovaries had eggs to much the same degree, leaving the myometrium and parametrium with the fewest eggs resident. Rectal tissue was relatively clean, surprizingly. Tissues of pelvic organs were positive for schistosome eggs in a total of 37: 33 by digestion technique, 24 by histology, and 20 by both. Inflammatory changes associated with schistosome eggs were found in 12 of 24 infections discovered histologically. Inflammation in the gential tract was only found in 3 cervixes and 2 vaginal walls; in bladders for example, infectious inflammation was seen in 7. Eggs of S. haematobium were identified in 19 of 24 histologically discovered infections. Eggs of S. haematobium were found in all 33 instances identified by the digestion technique, and in 9 of these instances, eggs of S. mansoni were also found. 7 of 9 infections with S. mansoni were of the rectum and 2 were of the bladder.^ieng

The effects of ethylene glycol monomethyl ether on testicular histology in F344 rats.

Ethylene glycol monomethyl ether (EGME) has been found to produce testicular atrophy in experimental rodents. The studies that follow were designed to determine the testicular cell type(s) most susceptible to EGME administration. For histologic studies, F344 rats were gavaged with 150 mg/kg/day of EGME 5 days per week, and serially sacrificed. In sections from perfusion-fixed tissue, necrotic changes were observed in some meiotic and premeiotic spermatocytes 24 hours after a single dose. Also, nuclear condensation was seen in occasional early pachytene spermatocytes. These effects were magnified after two doses; there were more necrotic pachytene and meiotic spermatocytes than necrotic stage I pachytene spermatocytes. By day 4, testes from all treated animals were affected; there was a pronounced maturation-depletion effect, seen as the absence of round spermatids from tubules in stages I to III. These effects continued to develop at days 7 and 10, leaving only Sertoli cells, spermatogonia, and late stage spermatids populating the epithelium. Other animals were treated similarly, but subject to efferent duct ligation 16 hours prior to sacrifice. Fluid production, as judged by weight gain in the testes after efferent duct ligation, was unaffected by EGME treatment. Analysis of the fluid collected at the rete testis indicated that there was no treatment-related change in the relative amounts of androgen binding protein. The data indicate that the spermatocyte is the primary target cell for the histologic effects of EGME in the testis of F344 rats.

Utricular otoconia of some amphibians have calcitic morphology.

This report concerns the morphological features of otoconia removed from the inner ear of four amphibian species. Results from scanning electron microscopic examination are compared based on the site of origin. These results show that utricular otoconia have a mineral structure that mimics calcite, rather than the widely accepted idea that they are mineralized by calcium carbonate of the aragonite polymorph.

Distribution and significance of norepinephrine in the lateral cochlear wall of pigmented and albino rats.

Picogram quantities of norepinephrine were found in cochlear regions of pigmented and non-pigmented rats. These regions of the cochlea were the modiolus, organ of Corti--osseous spiral lamina and the lateral cochlear wall. The content of norepinephrine in the modiolus and lateral cochlear wall of the pigmented rat was significantly greater than that in areas of the non-pigmented rat. In contrast, there was no statistical difference between the norepinephrine content of the organ of Corti--osseous spiral lamina region of the pigmented rat and that of the albino rat. Since a major difference between the pigmented and albino rats is the presence of melanin-containing melanocytes in the modiolus and lateral cochlear wall region of the pigmented animals, it is possible that norepinephrine is stored in cochlear melanocytes.

Quantitative evidence for cochlear, non-neuronal norepinephrine.

Endogenous norepinephrine was quantitatively measured in cochlear tissues of pigmented and nonpigmented animals by high-performance liquid chromatography and electrochemical detection. Epinephrine, dopamine and serotonin were not detected. The cochlear norepinephrine content of the pigmented animals was found to be more than double that present in corresponding albinos. Cochlear norepinephrine was only minimally depleted 48 h after surgical removal of the superior cervical ganglion. 12 h after administration of reserpine (5 mg/kg), cochlear norepinephrine was partially depleted. These results indicate that the norepinephrine located in the cochlea is probably not confined entirely to noradrenergic nerve terminals.

Synaptic changes in rat maculae in space and medical imaging: the link.

Two different space life sciences missions (SLS-1 and SLS-2) have demonstrated that the synapses of the hair cells of rat vestibular maculae increase significantly in microgravity. The results also indicate that macular synapses are sensitive to stress. These findings argue that vestibular maculae exhibit neuroplasticity to macroenvironmental and microenvironmental changes. This capability should be clinically relevant to rehabilitative training and/or pharmacological treatments for vestibular disease. The results of this ultrastructural research also demonstrated that type I and type II hair cells are integrated into the same neuronal circuitry. The findings were the basis for development of three-dimensional reconstruction software to learn details of macular wiring. This software, produced for scientific research, has now been adapted to reconstruct the face and skull directly from computerized tomography scans. In collaboration with craniofacial reconstructive surgeons at Stanford University Medical Center, an effort is under way to produce a virtual environment workbench for complex craniofacial surgery. When completed, the workbench will help surgeons train for and simulate surgery. The methods are patient specific. This research illustrates the value of basic research in leading to unanticipated medical applications.