Allozyme variation in a natural population of Stryphnodendron adstringens in the Rio Preto State Park, southeastern Brazil

Leaves and fruits from 63 Stryphnodendron adstringens trees were sampled in the Rio Preto State Park to analyze allozyme segregation, tissue specific expression of allozyme loci, and their genetic parameters. The enzyme systems ADH, EST, ACP, PGM, PGI, GDH, G6PDH, GOT, IDH, LAP, MDH, PER and SKDH were assessed by means of starch-gel electrophoresis. The polymorphic systems PGI, IDH, MDH and GOT demonstrated a dimeric quaternary structure, while EST and PER were monomeric. The total expected genetic diversity (H E) for leaves and seeds were 0.325 and 0.244 respectively. The effective number of alleles per locus (A E) was 1.58 in leaves and 1.42 in seeds. The values of H E and A E observed in S. adstringens were comparatively higher than the average values seen in allozyme studies of other woody plants. The values of the fixation indices for the population, considering leaves (f = 0.070) and seeds (f = 0.107), were not significant. The high values of genetic diversity and of effective number of alleles per locus, as well as the non-significant fixation index and the adjustments of the Hardy-Weinberg proportions between generations for the pgi-1, mdh-2 and idh-1 loci, indicated random mating in this population. The enzyme systems EST and PER demonstrated their best resolution in leaf tissues, while the MDH, IDH, PGI and GOT systems demonstrated their best resolution in seed tissues.

Estimates of genetic and phenotypic parameters in a segregant population of rice irrigated by continuous flooding

Estimates of genetic and phenotypic parameters were obtained by using data from families of a recurrent selection program in rice. An experiment using population CNA-IRAT 4ME/1/1 was conducted at two locations (Lambari and Cambuquira) in the State of Minas Gerais, Brazil. At Lambari, families S0:2 and S0:3 were assessed during crop seasons 1992/1993 and 1993/1994, respectively. In the Cambuquira trial, only S0:3 families were tested in 1993/1994. The experimental design was a 10 x 10 lattice with three replications. The following traits were assessed: grain yield (GY), mean number of days to flowering (FL), plant height (PH), and the incidence of neck blast (NB) caused by Pyricularia grisea and grain staining (GS) caused by Drechslera oryzae. This population proved to be promising for recurrent selection, as it had high average yield and genetic variability. Heritability estimates obtained using variance components were generally greater than estimates of realized heritability, and heritability obtained by parent-offspring regression

Predicting variety composite means without diallel crossing

Prediction of variety composite means was shown to be feasible without diallel crossing the parental varieties. Thus, the predicted mean for a quantitative trait of a composite is given by: Yk = a1 sigmaVj + a2sigmaTj + a3 - a4, with coefficients a1 = (n - 2k)/k²(n - 2); a2 = 2n(k - 1)/k²(n - 2); a3 = n(k - 1)/k(n - 1)(n - 2); and a4 = n²(k - 1)/k(n - 1)(n - 2); summation is for j = 1 to k, where k is the size of the composite (number of parental varieties of a particular composite) and n is the total number of parent varieties. Vj is the mean of varieties and Tj is the mean of topcrosses (pool of varieties as tester), and and are the respective average values in the whole set. Yield data from a 7 x 7 variety diallel cross were used for the variety means and for the "simulated" topcross means to illustrate the proposed procedure. The proposed prediction procedure was as effective as the prediction based on Yk = - ( -)/k, where and refer to the mean of hybrids (F1) and parental varieties, respectively, in a variety diallel cross. It was also shown in the analysis of variance that the total sum of squares due to treatments (varieties and topcrosses) can be orthogonally partitioned following the reduced model Yjj’ = mu + ½(v j + v j’) + + h j+ h j’, thus making possible an F test for varieties, average heterosis and variety heterosis. Least square estimates of these effects are also given

MHC-restricted antigen presentation and recognition: constraints on gene, recombinant and peptide vaccines in humans

The target of any immunization is to activate and expand lymphocyte clones with the desired recognition specificity and the necessary effector functions. In gene, recombinant and peptide vaccines, the immunogen is a single protein or a small assembly of epitopes from antigenic proteins. Since most immune responses against protein and peptide antigens are T-cell dependent, the molecular target of such vaccines is to generate at least 50-100 complexes between MHC molecule and the antigenic peptide per antigen-presenting cell, sensitizing a T cell population of appropriate clonal size and effector characteristics. Thus, the immunobiology of antigen recognition by T cells must be taken into account when designing new generation peptide- or gene-based vaccines. Since T cell recognition is MHC-restricted, and given the wide polymorphism of the different MHC molecules, distinct epitopes may be recognized by different individuals in the population. Therefore, the issue of whether immunization will be effective in inducing a protective immune response, covering the entire target population, becomes an important question. Many pathogens have evolved molecular mechanisms to escape recognition by the immune system by variation of antigenic protein sequences. In this short review, we will discuss the several concepts related to selection of amino acid sequences to be included in DNA and peptide vaccines.

Plasma metanephrines: a novel and cost-effective test for pheochromocytoma

Pheochromocytomas are rare chromaffin cell tumors that nevertheless must be excluded in large numbers of patients who develop sustained or episodic hypertension as well as in many others with suggestive symptoms or with a familial history of pheochromocytoma. Diagnosis of pheochromocytoma depends importantly on biochemical evidence of excess catecholamine production by a tumor. Imperfect sensitivity and specificity of commonly available biochemical tests and the low incidence of the tumor among the tested population mean that considerable time and effort can be expended in confirming or ruling out pheochromocytoma in patients where the tumor is suspected. Measurements of plasma free metanephrines provide a superior test compared to other available tests for diagnosis of pheochromocytoma. In particular, the high sensitivity of plasma free metanephrines means that a normal test result reliably excludes all but the smallest of pheochromocytomas so that no other tests are necessary. Measurements of plasma free metanephrines, when systematically combined with other diagnostic procedures outlined in this review, provide a more efficient, reliable and cost-effective approach for diagnosis of pheochromocytoma than offered by previously available approaches.

Effect of race, genetic population structure, and genetic models in two-locus association studies: clustering of functional renin-angiotensin system gene variants in hypertension association studies

Previous genetic association studies have overlooked the potential for biased results when analyzing different population structures in ethnically diverse populations. The purpose of the present study was to quantify this bias in two-locus association studies conducted on an admixtured urban population. We studied the genetic structure distribution of angiotensin-converting enzyme insertion/deletion (ACE I/D) and angiotensinogen methionine/threonine (M/T) polymorphisms in 382 subjects from three subgroups in a highly admixtured urban population. Group I included 150 white subjects; group II, 142 mulatto subjects, and group III, 90 black subjects. We conducted sample size simulation studies using these data in different genetic models of gene action and interaction and used genetic distance calculation algorithms to help determine the population structure for the studied loci. Our results showed a statistically different population structure distribution of both ACE I/D (P = 0.02, OR = 1.56, 95% CI = 1.05-2.33 for the D allele, white versus black subgroup) and angiotensinogen M/T polymorphism (P = 0.007, OR = 1.71, 95% CI = 1.14-2.58 for the T allele, white versus black subgroup). Different sample sizes are predicted to be determinant of the power to detect a given genotypic association with a particular phenotype when conducting two-locus association studies in admixtured populations. In addition, the postulated genetic model is also a major determinant of the power to detect any association in a given sample size. The present simulation study helped to demonstrate the complex interrelation among ethnicity, power of the association, and the postulated genetic model of action of a particular allele in the context of clustering studies. This information is essential for the correct planning and interpretation of future association studies conducted on this population.

Cost-effective analysis of different algorithms for the diagnosis of hepatitis C virus infection

We compared the cost-benefit of two algorithms, recently proposed by the Centers for Disease Control and Prevention, USA, with the conventional one, the most appropriate for the diagnosis of hepatitis C virus (HCV) infection in the Brazilian population. Serum samples were obtained from 517 ELISA-positive or -inconclusive blood donors who had returned to Fundação Pró-Sangue/Hemocentro de São Paulo to confirm previous results. Algorithm A was based on signal-to-cut-off (s/co) ratio of ELISA anti-HCV samples that show s/co ratio ³95% concordance with immunoblot (IB) positivity. For algorithm B, reflex nucleic acid amplification testing by PCR was required for ELISA-positive or -inconclusive samples and IB for PCR-negative samples. For algorithm C, all positive or inconclusive ELISA samples were submitted to IB. We observed a similar rate of positive results with the three algorithms: 287, 287, and 285 for A, B, and C, respectively, and 283 were concordant with one another. Indeterminate results from algorithms A and C were elucidated by PCR (expanded algorithm) which detected two more positive samples. The estimated cost of algorithms A and B was US$21,299.39 and US$32,397.40, respectively, which were 43.5 and 14.0% more economic than C (US$37,673.79). The cost can vary according to the technique used. We conclude that both algorithms A and B are suitable for diagnosing HCV infection in the Brazilian population. Furthermore, algorithm A is the more practical and economical one since it requires supplemental tests for only 54% of the samples. Algorithm B provides early information about the presence of viremia.

Synaptic vesicle pool size, release probability and synaptic depression are sensitive to Ca2+ buffering capacity in the developing rat calyx of Held

The calyx of Held, a specialized synaptic terminal in the medial nucleus of the trapezoid body, undergoes a series of changes during postnatal development that prepares this synapse for reliable high frequency firing. These changes reduce short-term synaptic depression during tetanic stimulation and thereby prevent action potential failures during a stimulus train. We measured presynaptic membrane capacitance changes in calyces from young postnatal day 5-7 (p5-7) or older (p10-12) rat pups to examine the effect of calcium buffer capacity on vesicle pool size and the efficiency of exocytosis. Vesicle pool size was sensitive to the choice and concentration of exogenous Ca2+ buffer, and this sensitivity was much stronger in younger animals. Pool size and exocytosis efficiency in p5-7 calyces were depressed by 0.2 mM EGTA to a greater extent than with 0.05 mM BAPTA, even though BAPTA is a 100-fold faster Ca2+ buffer. However, this was not the case for p10-12 calyces. With 5 mM EGTA, exocytosis efficiency was reduced to a much larger extent in young calyces compared to older calyces. Depression of exocytosis using pairs of 10-ms depolarizations was reduced by 0.2 mM EGTA compared to 0.05 mM BAPTA to a similar extent in both age groups. These results indicate a developmentally regulated heterogeneity in the sensitivity of different vesicle pools to Ca2+ buffer capacity. We propose that, during development, a population of vesicles that are tightly coupled to Ca2+ channels expands at the expense of vesicles more distant from Ca2+ channels.

Characterization of alpha thalassemic genotypes by multiplex ligation-dependent probe amplification in the Brazilian population

Alpha-thalassemia is the most common inherited disorder of hemoglobin synthesis. Genomic deletions involving the alpha-globin gene cluster on chromosome 16p13.3 are the most frequent molecular causes of the disease. Although common deletions can be detected by a single multiplex gap-PCR, the rare and novel deletions depend on more laborious techniques for their identification. The multiplex ligation-dependent probe amplification (MLPA) technique has recently been used for this purpose and was successfully used in the present study to detect the molecular alterations responsible for the alpha-thalassemic phenotypes in 8 unrelated individuals (3 males and 5 females; age, 4 months to 30 years) in whom the molecular basis of the disease could not be determined by conventional methods. A total of 44 probe pairs were used for MLPA, covering approximately 800 kb from the telomere to the MSLN gene in the 16p13.3 region. Eight deletions were detected. Four of these varied in size from 240 to 720 kb and affected a large region including the entire alpha-globin gene cluster and its upstream regulatory element (alpha-MRE), while the other four varied in size from 0.4 to 100 kb and were limited to a region containing this element. This study is the first in Brazil to use the MLPA method to determine the molecular basis of alpha-thalassemia. The variety of rearrangements identified highlights the need to investigate all cases presenting microcytosis and hypochromia, but without iron deficiency or elevated hemoglobin A2 levels and suggests that these rearrangements may be more frequent in our population than previously estimated.

A morphometric study of the lumbar spinous process in the Chinese population

Our goal was to analyze the anatomical parameters of the lumbar spine spinous process for an interspinous stabilization device designed for the Chinese population and to offer an anatomical basis for its clinical application. The posterior lumbar spines (T12-S1) of 52 adult cadavers were used for measuring the following: distance between two adjacent spinous processes (DB), distance across two adjacent spinous processes (DA), thickness of the central spinous processes (TC), thickness of the superior margin of the spinous processes (TS), thickness of the inferior margin of the spinous processes (TI), and height of the spinous processes (H). Variance and correlation analyses were conducted for these data, and the data met the normal distribution and homogeneity of variance. DB decreased gradually from L1-2 to L5-S1. DA increased from T12-L1 to L2-3 and then decreased from L2-3 to L4-5. The largest H in males was noted at L3 (25.45±5.96 mm), whereas for females the largest H was noted at L4 (18.71±4.50 mm). Usually, TS of the adjacent spinous process was lower than TI. Based on the anatomical parameters of the lumbar spinous processes obtained in this study, an “H”-shaped coronal plane (posterior view) was proposed as an interspinous stabilization device for the Chinese population. This study reports morphometric data of the lumbar spinous processes in the Chinese population, which provides an anatomical basis for future clinical applications.

Exploring the experiences of parents with children who have communication delays, as a population at risk for the development of behavioural concerns

Existing research identifies preschoolers with communication delays as a population at risk for the development of behavioural concerns. This risk increases when additional environmental factors such as parental stress and family conflict are also present. Research has also shown that behavioural concerns can be stable over time when they develop early. However, early intervention has been shown to be effective in addressing these concerns. The effectiveness of early intervention in addressing both child and family outcomes increases when interventions are delivered in a family-centred approach. This research project made use of data related to child behaviour and parenting, gathered through the Family Resource Project which explored the parenting experiences and resource access and allocation decisions of families who have preschool children with and without communication delays. Cluster analysis was used to explore whether there were identifiable clusters of children and families within each sample. Interview data fi"om each identified family cluster was then explored further, to identify how parents described their child's behaviour and their experience of parenting. Results show that, within this sample, parents of preschoolers with communication delays described their child's behaviour and their experience of parenting differently than did parents of children without communication delays. Results also showed that within this sample parents experiencing parental stress and/or family conflict described their child's behaviour and their experience of parenting differently than did parents from other clusters. Results suggest support for early intervention and the use of family-centred intervention, particularly for families of children with communication delays.

The effectiveness of a health versus appearance message on pregnant women's intentions to exercise postpartum : the moderating role of self-monitoring

Despite the confimied health benefits of exercise during the postpartum period, many new mothers are not sufficiently active. The present research aimed to examine the effectiveness of 2 types of messages on intention to exercise after giving birth on 2 groups of pregnant women (low and high self-monitors) using the Theory of Planned Behavior as a theoretical basis. Participants were 2 1 8 pregnant women 1 8 years of age and older (Mean age = 27.9 years, SD = 5.47), and in their second or third trimester. Women completed a demographics questionnaire, a self-monitoring (SM) scale and the Godin Leisure Time Exercise Questionnaire for current and pre-pregnancy exercise levels. They then read one of two brochures, describing either the health or appearance benefits of exercise for postpartum women. Women's attitudes, social norms, perceived behavioral control, and intentions to exercise postpartum were then assessed to determine whether one type of message (health or appearance) was more effective for each group. A MANOVA found no significant effect (p>0.05) for message type, SM, or their interaction. Possible reasons include the fact that the two messages may have been too similar, reading any message about exercise may result in intentions to exercise, or lack of attention given to the brochure. Given the lack of research in this area, more studies are necessary to confirm the present results. Two additional exploratory analyses were conducted. Pearson correlations found higher levels of pre-pregnancy exercise and current exercise to be associated with more positive attitudes, more positive subjective norms, higher perceived behavioral control, and higher intention to exercise postpartum. A hierarchical regression was conducted to determine the predictive utility of attitudes, subjective norms, and perceived behavioral control on intention for each self-monitoring group. Results of the analysis demonstrated the three independent variables significantly predicted intention (p < .001) in both groups, accounting for 58-62% of the variance in intention. For low self-monitors, attitude was the strongest predictor of intention, followed by perceived behavioral control and subjective norm. For high self-monitors, perceived behavioral control was the strongest predictors, followed by attitudes and subjective norm. The present study has practical and real world implications by contributing to our understanding of what types of messages, in a brochure format, are most effective in changing pregnant women's attitudes, subjective norm, perceived behavioral control and intention to exercise postpartum and provides ftirther support for the use of the Theory of Planned Behavior with this population.

Differentiation of morphology and reproductive traits in the house finch (Carpodacus mexicanus): a comparison between native and introduced populations

House Finches (CarpQdacqs mexiCAnuS) were introduced to Long Island, New York from southern'California in 1940. Apparently, an initial sample of less than 100 birds has given rise to a population that now occupies much of the eastern United States. This study was to determine if morphological and reproductive changes have taken place in introduced eastern birds, which have colonized a novel environment. A study area in Goleta, California (CAL) represented the parental population whereas for comparison, House Finches in St. Catharines, Ontario (ONT) represented the introduced population. Interlocality variation in 25 morphometric characters of 100 adult House Finches was examined statistically. Singleclassification analysis of variance revealed significant interlocality differentiation in seven characters of males and nine of females. Females showed differentiation in more limb elements than males. Analysis of character variation using discriminant and principal component analysis distinguished samples on the basis of variation in shape. Compared to CAL, aNT birds (especially females) had smaller extremities relative to certain core parts and weight. Females showed similar patterns of character covariation in each locality on the second principal component, which suggests that differentiation of the ONT population may not be solely environmentally induced. Sexual dimorphism was evident in four charaoters in aNT and five in CAL. Disoriminant analysis distinguished sex on the basis of variation in shape. Males possessed a relatively larger flying apparatus and small.er hind limbs than females. The dearee of sexual dimorphism did not vary sicnifioantly between looalities. 3 Data on reproduotive parameters were oolleoted in 1983 and 1984 in ONT, and 1984 in CAL. In 1984, Bouse Finohes began breedina approximately three months earlier in CAL than in ONT. In ONT, there was no sianifioant differenoe in mean olutoh initiation date between 1983 and 1984. In both looalities most nests oontained either four or five ea",s, and olutoh size differenoes between looalites were not signifioant. Seasonal deolines in olutch size were evident in ONT but not in CAL. Intralooality variation in e.g weight and size was not related to clutch size. E",g weiaht showed no seasonal trend in ONT, but inoreased sianifioantly with breed ina season in OAL. In both looalities e8'''' weiaht increased sipifioantly with order of layina in olutohes of four but not in clutohes of five. Eag's in ONT in 1983 and 1984 were sip.ificantly larser than in CAL in 1984. The modal inoubation period was 13 days and did not vary sip.ifioantly between localites. In both looalities nestling weiaht on the day of hatohing was oorrelated to fresh ega welaht. For muoh of the period between hatohing and 14 days post-hatoh, ONT nestlinas were signifioantly laraer than CAL nestlings in terms of weiaht. bill length, bill depth, and manus length.

Size regulation of double and half embryos in the mice musculus

Genetic chimeras made by aggregating early mouse embryos have many uses in developmental biology and have also provided insights into embryonic growth regulation. There is an indication that the embryo can regulate for an increase in size because although aggregation chimeras are twice as big as normal embryos when made, they are born of normal size. Upward regula..... tion of size reduced embryos is also possible. Half embryos made by the isolation or destruction of one of the blastomeres of a 2-cell embryo are also born of normal size. Little is known about the timing or the mechanism of this size regulation. In this study, the timing of size regulation in double and half embryos was clearly established by comparison of cell numbers derived from serial reconstruction of light microscope sections of control and experimental embryos. It was shown that size regulation in double embryos occurred around 6dl6h and in half embryos by 7dOh. Size regulation occurred in all tissues at the same time indicating a single control mechanism for the entire embryo. More detailed examination of the growth of double embryos revealed that size regulation occurred by alteration in cell cycle length~ No excessive cell death was found in double embryos compared to the controls and continuous labelling with [3H] thymidine showed no large non-dividing cell population in double embryos. However, a comparison of the mitotic index of double and control embryos after colcemid treatment, revealed a large difference between the two around 5dl6h to 6d16h. During this period, control embryos underwent a proliferative burst not shown by the double embryos. The mechanism for cell cycle control is not clear; it may be intrinsic to the embryo or determined by the uterine environment. Evidence was found suggesting that differentiation in the postimplantation embryo was cell number dependent. The timing of differentiative events was examined in half, double and control embryos. Proamnion formation, which occurs prior to size regulation, occurs at the same cell number but at different times in the three groups of embryos. However mesoderm which appears after size regulation was seen at the same time in all grollps of embryos.

Indices and Implications of Emotional Underarousal for Persons with a History of Head Trauma

We examined the role of altered emotional functioning across the spectrum of injury severity (mild head injury [MHI], moderate/severe traumatic brain injury [TBI]), its implications for social behaviours, and the effect of modifying arousal and its relation to cognitive performance. In the first study (N = 230), students with self-reported MHI endorsed engaging in socially unacceptable and erratic behaviours significantly more often than did those with no MHI. We did not find significant differences between the groups in the measure of emotional intelligence (EI); however, for students who reported a MHI, scores on the EI measure significantly predicted reports of socially unacceptable behaviours such that lower scores predicted poorer social functioning, accounting for approximately 20% of the variance. Also, the experience of postconcussive symptoms was found to be significantly greater for students with MHI relative to their peers. In the second study (N = 85), we further examined emotional underarousal in terms of physiological (i.e., electrodermal activation [EDA]) and self-reported responsivity to emotionally-evocative picture stimuli. Although the valence ratings of the stimuli did not differ between students with and without MHI as we had expected, we found evidence of reduced and/or indiscriminate emotional responding to the stimuli for those with MHI which mimics that observed in other studies for persons with moderate/severe TBI. We also found that emotional underarousal followed a gradient of injury severity despite reporting a pattern of experiencing more life stressors. In the third study (N = 81), we replicated our findings of emotional underarousal for those with head trauma and also uniquely explored neuroendocrine aspects (salivary cortisol; cortisol awakening response [CAR]) and autonomic indices (EDA) of emotional dysregulation in terms of stress responsivity across the spectrum of injury severity (MHI [n = 32], moderate/severe TBI [n = 9], and age and education matched controls [n = 40]). Although the manipulation was effective in modifying arousal state in terms of autonomic and self-reported indices, we did not support our hypothesis that increased arousal would be related to improved performance on cognitive measures for those with prior injury. To our knowledge, this is the only study to examine the CAR with this population. Repeated measure analysis revealed that, upon awakening, students with no reported head trauma illustrated the typical CAR increase 45 minutes after waking, whereas, students who had a history of either mild head trauma or moderate/severe TBI demonstrated a blunted CAR. Thus, across the three studies we have provided evidence of emotional underarousal, its potential implications for social interactions, and also have identified potentially useful indices of dysregulated stress responsivity regardless of injury severity.