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Abstract Background Next generation sequencing technologies often require numerous primer designs that require good target coverage that can be financially costly. We aimed to develop a system that would implement primer reuse to design degenerate primers that could be designed around SNPs, thus find the fewest necessary primers and the lowest cost whilst maintaining an acceptable coverage and provide a cost effective solution. We have implemented Metropolis-Hastings Markov Chain Monte Carlo for optimizing primer reuse. We call it the Markov Chain Monte Carlo Optimized Degenerate Primer Reuse (MCMC-ODPR) algorithm. Results After repeating the program 1020 times to assess the variance, an average of 17.14% fewer primers were found to be necessary using MCMC-ODPR for an equivalent coverage without implementing primer reuse. The algorithm was able to reuse primers up to five times. We compared MCMC-ODPR with single sequence primer design programs Primer3 and Primer-BLAST and achieved a lower primer cost per amplicon base covered of 0.21 and 0.19 and 0.18 primer nucleotides on three separate gene sequences, respectively. With multiple sequences, MCMC-ODPR achieved a lower cost per base covered of 0.19 than programs BatchPrimer3 and PAMPS, which achieved 0.25 and 0.64 primer nucleotides, respectively. Conclusions MCMC-ODPR is a useful tool for designing primers at various melting temperatures at good target coverage. By combining degeneracy with optimal primer reuse the user may increase coverage of sequences amplified by the designed primers at significantly lower costs. Our analyses showed that overall MCMC-ODPR outperformed the other primer-design programs in our study in terms of cost per covered base.

Abstract Background To study the dynamics of wild-type and drug-resistant HIV-1 RT variants, we developed a methodology that follows the fates of individual genomes over time within the viral quasispecies. Single genome sequences were obtained from 3 pigtail macaques infected with a recombinant simian immunodeficiency virus containing the RT coding region from HIV-1 (RT-SHIV) and treated with short-course efavirenz monotherapy 13 weeks post-infection followed by daily combination antiretroviral therapy (ART) beginning at week 17. Bioinformatics tools were constructed to trace individual genomes from the beginning of infection to the end of the treatment. Results A well characterized challenge RT-SHIV inoculum was used to infect three monkeys. The RT-SHIV inoculum had 9 variant subpopulations and the dominant subpopulation accounted for 80% of the total genomes. In two of the three monkeys, the inoculated wild-type virus was rapidly replaced by new wild type variants. By week 13, the original dominant subpopulation in the inoculum was replaced by new dominant subpopulations, followed by emergence of variants carrying known NNRTI resistance mutations. However, during ART, virus subpopulations containing resistance mutations did not outgrow the wide-type subpopulations until a minor subpopulation carrying linked drug resistance mutations (K103N/M184I) emerged. We observed that persistent viremia during ART is primarily made up of wild type subpopulations. We also found that subpopulations carrying the V75L mutation, not known to be associated with NNRTI resistance, emerged initially in week 13 in two macaques. Eventually, all subpopulations from these two macaques carried the V75L mutation. Conclusion This study quantitatively describes virus evolution and population dynamics patterns in an animal model. The fact that wild type subpopulations remained as dominant subpopulations during ART treatment suggests that the presence or absence of at least some known drug resistant mutations may not greatly affect virus replication capacity in vivo. Additionally, the emergence and prevalence of V75L indicates that this mutation may provide the virus a selective advantage, perhaps escaping the host immure system surveillance. Our new method to quantitatively analyze viral population dynamics enabled us to observe the relative competitiveness and adaption of different viral variants and provided a valuable tool for studying HIV subpopulation emergence, persistence, and decline during ART.

Abstract Background Recent pathological studies have suggested that thalamic degeneration may represent a site of non-dopaminergic degeneration in Parkinson's Disease (PD). Our objective was to determine if changes in the thalami could be non-invasively detected in structural MRI images obtained from subjects with Parkinson disease (PD), compared to age-matched controls. Results No significant differences in volume were detected in the thalami between eighteen normal subjects and eighteen PD subjects groups. However significant (p < 0.03) shape differences were detected between the Left vs. Right thalami in PD, between the left thalami in PD and controls, and between the right thalami in PD and controls using a recently-developed, spherical harmonic-based representation. Conclusion Systematic changes in thalamic shape can be non-invasively assessed in PD in vivo. Shape changes, in addition to volume changes, may represent a new avenue to assess the progress of neurodegenerative processes. Although not directly discernable at the resolution of standard MRI, previous pathological studies would suggest that the shape changes detected in this study represent degeneration in the centre median-parafascicular (CM-Pf) complex, an area known to represent selective non-dopaminergic degeneration in PD.

The family of pro-inflammatory and pro-angiogenic chemokines including Interleukin-8 (IL-8, aka CXCL8) and its homologues (CXCL1,2,3,5,6, and 7) exhibit promiscuous binding and activation of several G-protein-coupled receptors (i.e., CXCR2, CXCR1, and the atypical chemokine receptor (ACKR1)). A high proportion of their biological activity is attributed to CXCR2 activation, thus many CXCR2 inhibitors are in clinical trials for several chronic diseases. However, CXCR2 inhibition is often only investigated acutely in these trials or in Cxcr2 -/- mice grown in gnotobiotic conditions. Since humans do not live in germ-free environments, our first goal is to highlight novel retinal and systemic observations in Cxcr2 -/- mice grown in non-gnotobiotic conditions that suggest potential harmful consequences of long-term CXCR2 deficiency or blockade. Beyond confirmation of circulating blood/immune cell-related phenotypes, we report novel findings in Cxcr2 -/- mice including: (1) delayed dye transit to the retinal vasculature, (2) alterations in the density and distribution of retinal vessels, astrocytes and microglia, (3) decreased electroretinogram a- and b-wave amplitudes, (4) reduced visual acuity, and (5) increased polymorphonuclear cell accumulation in vascular lumina abutting venular walls in the retina and in vital non-ocular tissues (lung and liver). Furthermore, PheWAS of CXCR2 CXCR1, and ACKR1 gene variants using data from UK Biobank participants suggest clinical associations with both retinal and vascular disease phenotypes. We conclude that chronic CXCR2 deficiency in mice contributes to functional damage to the retina and that the long-term safety of CXCR1/2 inhibitors designed for chronic use in humans should be explored before clinical adoption to safeguard sight and overall vascular health., (© 2024. The Author(s).)

Purpose: The purpose of this study was to examine comparability between 2 editions of the Test of Gross Motor Development (TGMD-second and TGMD-third edition) on (1) how children's motor skills were categorized as average or below average, and (2) how children are screened for being at-risk for motor delay or with delayed motor skills., Methods: Participants were 226 children (Mage = 53.4 mo, 125 boys). All children completed full TGMD-2 and TGMD-3. Children were classified as average or above (>25th percentile) or below average (≤25th percentile) and, when applicable, as developmental delay (≤5th percentile) or at-risk for developmental delay (6-25th percentile). We compared children's classifications across TGMD editions using percent agreement and chi-squared tests., Results: The TGMD-2 and TGMD-3 had moderate agreement when categorizing children as below average (72.2% for total skills, 76.0% for locomotor skills, and 73% for ball skills). The TGMD-3 was significantly more likely to categorize children's motor skill performance as average or above (all P < .01)., Conclusion: TGMD-2 and TGMD-3 similarly screen children who demonstrate below average skills (≤25th percentile), but not for specific skill level classifications, including above average, at-risk for delays, and delayed.

Ocular levels of IL-1β, TNFα, IL-8, and IL-6 correlate with progression of diabetic retinopathy (DR). Müller cells (MC), which are crucial to maintaining retinal homeostasis, are targets and sources of these cytokines. We explored the relative capacities of these four DR-associated cytokines to amplify inflammatory signal expression both in and between human MC (hMC) and retinal microvascular endothelial cells (hRMEC) and in the mouse retina. Of the four cytokines, IL-1β was the most potent stimulus of transcriptomic alterations in hMC and hRMEC in vitro, as well as in the mouse retina after intravitreal injection in vivo. Stimulation with IL-1β significantly induced expression of all four transcripts in hMC and hRMEC. TNFα significantly induced expression of some, but not all, of the four transcripts in each cell, while neither IL-8 nor IL-6 showed significant induction in either cell. Similarly, conditioned media (CM) derived from hMC or hRMEC treated with IL-1β, but not TNFα, upregulated inflammatory cytokine transcripts in the reciprocal cell type. hRMEC responses to hMC-derived CM were dependent on IL-1R activation. In addition, we observed a correlation between cytokine expression changes following direct and CM stimulation and NFκB-p65 nuclear translocation in both hMC and hRMEC. Finally, in mice, intravitreal injections of IL-1β, but not TNFα, induced retinal expression of Il1b and CXCL8 homologues Cxcl1, Cxcl2, Cxcl3, and Cxcl5, encoding pro-angiogenic chemokines. Our results suggest that expression of IL-1β, TNFα, IL-8, and IL-6 may be initiated, propagated, and sustained by autocrine and paracrine signals in hRMEC and hMC through a process involving IL-1β and NFκB. Targeting these signals may help thwart inflammatory amplification, preventing progression to vision-threatening stages and preserving sight., Competing Interests: Declaration of competing interest There are no financial or non-financial competing interests declared by any author., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Regulatory T cells (T regs ) are protective in atherosclerosis but reduced during disease progression due to cell death and loss of stability. However, the mechanisms of T reg dysfunction remain unknown. Oxidized phospholipids are abundant in atherosclerosis and can activate innate immune cells, but little is known regarding their impact on T cells. Given T reg loss during atherosclerosis progression and oxidized phospholipid levels in the plaque microenvironment, we investigated whether oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (oxPAPC), an oxidized phospholipid associated with atherosclerotic plaques, alters T reg differentiation and function., Methods: CD4 + T cells were polarized to T reg , T helper (Th) 1, and Th17 cells with or without oxPAPC and assessed by flow cytometry. Gene expression in oxPAPC-treated T regs was analyzed by bulk RNA sequencing. Functional studies of oxPAPC-induced T regs were performed by coculturing T regs with CellTrace Violet-labeled cells in vitro, and by adoptively transferring T regs to hyperlipidemic Ldlr -/- mice to measure atherosclerosis progression., Results: Compared with controls, oxPAPC-treated T regs were less viable, but surviving cells expressed higher levels of the Th1-associated markers T-bet, CXCR3, and IFN (interferon)-γ. Th1 and Th17 skewing cultures were unaltered by oxPAPC. IFN-γ is linked to T reg instability, thus T reg polarization experiments were repeated using Ifngr1 -/- CD4 + T cells. IFNγR1 (INF gamma receptor 1) deficiency did not improve cell viability in oxPAPC-treated T regs ; however, T-bet and IFN-γ expression was not increased in surviving cells suggesting a role for IFN-γsignaling. OxPAPC-treated T regs were less suppressive in vitro, and adoptive transfer studies in hyperlipidemic Ldlr -/- mice showed that oxPAPC-induced T regs possessed altered tissue homing and were insufficient to inhibit atherosclerosis progression., Conclusions: OxPAPC elicits T reg -specific changes altering T reg differentiation and inducing a Th1-like phenotype in surviving cells partially through IFN-γ signaling. This is biologically relevant as oxPAPC-treated T regs do not reduce atherosclerosis progression in Ldlr -/- mice. This study supports the role of oxidized phospholipids in negatively impacting T reg differentiation and atheroprotective function., Competing Interests: Disclosures None.

This study examined the effects of a motor-skill intervention on children's perceived motor competence (PMC; object control, locomotor, and combined [total]) and explored if effects differed between the sexes. Preschoolers (N = 274; 47.96 months) completed either a motor-skill intervention (the Children's Health Activity Motor Program [CHAMP]) or recess. PMC was measured with the Digital Scale of PMC before and after each condition. Controlling for pretest scores, recess girls had lower posttest object-control PMC scores than CHAMP boys, CHAMP girls, and recess boys (all p < .05). CHAMP children had significantly higher posttest locomotor and total PMC (all p < .001) compared with children who engaged in recess. CHAMP partially eliminates sex differences in PMC, particularly for object-control skills. Girls who participated in recess did not increase PMC like children in CHAMP and boys who engaged in outdoor recess.

It is well-documented that intelligence quotient (IQ) is a poor predictor of adaptive behavior scores in autism, with autistic children having lower adaptive behavior scores than would be predicted based on their IQ scores. Differences in motor skills may explain the variability in their adaptive behavior scores. The current study examined how motor skills might explain autistic individuals' low adaptive behavior scores and which individual components of IQ (i.e., verbal comprehension and perceptual reasoning) and motor skills (i.e., manual dexterity, aiming and catching, and balance) may drive this effect. We examined the associations between IQ, motor skills, calibrated severity, and adaptive behavior scores in 45 autistic children and adolescents. Using a t-test, we found a significant difference (p <0.001) between full-scale IQ and adaptive behavior scores, indicating that our participants' adaptive behavior scores were lower than would be expected given their full-scale IQ. Using a linear regression, we investigated whether motor skills predicted adaptive behavior in autistic children and adolescents and found that motor skills scores were associated with adaptive behavior scores (p = 0.022). To further investigate these associations, we used another linear regression to examine how individual components of IQ and motor skills predicted adaptive behavior scores in autistic children and adolescents. Our results indicated that manual dexterity scores were associated with adaptive behavior scores (p = 0.036). These findings clearly illustrate the need for further understanding of autistic individuals' difficulties with adaptive behavior and the potential role of motor skill difficulties that may underlie these difficulties. LAY SUMMARY: Autistic children have lower adaptive behavior scores (e.g., daily living skills, social skills, communication) than intelligence scores (e.g., verbal and perceptual skills) along with difficulties with motor skills. Motor skills may explain the gap between adaptive behavior and intelligence. We found motor skills were associated with adaptive behavior in autistic children and adolescents. In particular, hand coordination was associated with adaptive behavior. We need to better understand how autistic individuals' motor skills impact their adaptive behavior to provide effective supports., (© 2022 The Authors. Autism Research published by International Society for Autism Research and Wiley Periodicals LLC.)

The recovery of ancient RNA from archeological material could enable the direct study of microevolutionary processes. Small RNAs are a rich source of information because their small size is compatible with biomolecular preservation, and their roles in gene regulation make them likely foci of evolutionary change. We present here the small RNA fraction from a sample of archeological barley generated using high-throughput sequencing that has previously been associated with localized adaptation to drought. Its microRNA profile is broadly similar to 19 globally distributed modern barley samples with the exception of three microRNAs (miRNA159, miRNA319, and miR396), all of which are known to have variable expression under stress conditions. We also found retrotransposon activity to be significantly reduced in the archeological barley compared with the controls, where one would expect the opposite under stress conditions. We suggest that the archeological barley's conflicting stress signals could be the result of long-term adaptation to its local environment., (© The Author 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.)

Our understanding of the evolution of domestication has changed radically in the past 10 years, from a relatively simplistic rapid origin scenario to a protracted complex process in which plants adapted to the human environment. The adaptation of plants continued as the human environment changed with the expansion of agriculture from its centres of origin. Using archaeogenomics and computational models, we can observe genome evolution directly and understand how plants adapted to the human environment and the regional conditions to which agriculture expanded. We have applied various archaeogenomics approaches as exemplars to study local adaptation of barley to drought resistance at Qasr Ibrim, Egypt. We show the utility of DNA capture, ancient RNA, methylation patterns and DNA from charred remains of archaeobotanical samples from low latitudes where preservation conditions restrict ancient DNA research to within a Holocene timescale. The genomic level of analyses that is now possible, and the complexity of the evolutionary process of local adaptation means that plant studies are set to move to the genome level, and account for the interaction of genes under selection in systems-level approaches. This way we can understand how plants adapted during the expansion of agriculture across many latitudes with rapidity.

The purpose of this study was to perform a systematic review to determine if respiratory muscle training (RMT) improves sport performance and respiratory muscle strength and endurance. Methodology followed the Cochrane Collaboration protocol. MEDLINE, CINAHL, SPORTDiscus, PEDro, EMBASE, EBM reviews, and COCHRANE electronic databases were searched until July 2011. Articles were included if: (a) participants were athletes; (b) RMT was compared with sham or control in a randomized controlled design and included outcomes of respiratory muscle and sport performance; and (d) published in English. Quality assessment using PEDro and data abstraction was performed by 2 authors. Outcomes evaluated were measures of sport performance, exercise capacity, spirometry, and respiratory muscle strength and endurance. Meta-analyses were performed on outcomes reported in 2 or more papers. Results of this systematic review revealed that of the 6,923 citations retrieved from the search strategy, 21 met the inclusion criteria. Meta-analyses demonstrated a significant positive effect of RMT on sport performance outcomes of time trials, exercise endurance time, and repetitions on Yo-Yo tests. Inspiratory muscle strength and endurance improved in most studies, which in part, was dependent on the type of RMT employed. Determination of the type of athlete that may benefit most from RMT was limited by small sample sizes, differing RMT protocols, and differences in outcome measures across studies. In conclusion, RMT can improve sport performance. Closer attention to matching the ventilatory demands of RMT to those required during athletic competition and more aggressive progression of training intensity may show greater improvements in future studies.

Transposable elements (TEs) are drivers of evolution resulting in episodic surges of genetic innovation and genomic reorganization (Oliver KR, Greene WK. 2009. TEs: powerful facilitators of evolution. Bioessays 31:703-714.), but there is little evidence of the timescale in which this process has occurred (Gingerich PD. 2009. Rates of evolution. Ann Rev Ecol Evol Syst. 40:657-675.). The paleontological and archaeological records provide direct evidence for how evolution has proceeded in the past, which can be accessed through ancient DNA to examine genomes using high-throughput sequencing technologies (Palmer SA, Smith O, Allaby RG. 2011. The blossoming of plant archaeogenetics. Ann Anat. 194:146-156.). In this study, we report shotgun sequencing of four archaeological samples of cotton using the GS 454 FLX platform, which enabled reconstruction of the TE composition of these past genomes and species identification. From this, a picture of lineage specific evolutionary patterns emerged, even over the relatively short timescale of a few thousand years. Genomic stability was observed between South American Gossypium barbadense samples separated by over 2,000 miles and 3,000 years. In contrast, the TE composition of ancient Nubian cotton, identified as G. herbaceum, differed dramatically from that of modern G. herbaceum and resembled closely the A genome of the New World tetraploids. Our analysis has directly shown that considerable genomic reorganization has occurred within the history of a domesticated plant species while genomic stability has occurred in closely related species. A pattern of episodes of rapid change and periods of stability is expected of punctuated evolution. This observation is important to understanding the process of evolution under domestication.

Recent years have seen a broadening range of studies undertaken in the field of plant archaeogenetics as the field has entered a new stage of maturity. There has been a movement towards a more functional understanding of plant evolution that has been facilitated by sequencing technologies and increasingly powerful genomic annotation. Studies have progressed to consideration of genes of pertinent function, metagenomics and investigation of the genetic basis of domestication traits coupled with human agricultural selection. Recent applications of high-throughput sequencing have facilitated archaeogenomic investigations and allowed increasingly fragmented DNA to be retrieved and characterised. While these developments are leading to the selective resequencing of targeted genomic regions in archaeobotanical samples, they have also opened up new avenues of opportunity in terms of the accessibility of biomolecules. Material once thought to be void of useful quantities of ancient DNA is now being accessed in the light of diagenetic information thrown up by HTS. The amount of archaeobotanical material that can be utilised and the work that is possible with that material are both increasing making it likely that a fertile period of plant archaeogenetics is underway., (Copyright © 2011. Published by Elsevier GmbH.)

Background: Archaeobotanical samples of barley (Hordeum vulgare L.) found at Qasr Ibrim display a two-row phenotype that is unique to the region of archaeological sites upriver of the first cataract of the Nile, characterised by the development of distinctive lateral bracts. The phenotype occurs throughout all strata at Qasr Ibrim, which range in age from 3000 to a few hundred years., Methodology and Findings: We extracted ancient DNA from barley samples from the entire range of occupancy of the site, and studied the Vrs1 gene responsible for row number in extant barley. Surprisingly, we found a discord between the genotype and phenotype in all samples; all the barley had a genotype consistent with the six-row condition. These results indicate a six-row ancestry for the Qasr Ibrim barley, followed by a reassertion of the two-row condition. Modelling demonstrates that this sequence of evolutionary events requires a strong selection pressure., Conclusions: The two-row phenotype at Qasr Ibrim is caused by a different mechanism to that in extant barley. The strength of selection required for this mechanism to prevail indicates that the barley became locally adapted in the region in response to a local selection pressure. The consistency of the genotype/phenotype discord over time supports a scenario of adoption of this barley type by successive cultures, rather than the importation of new barley varieties associated with individual cultures.

The reference sequence for each human chromosome provides the framework for understanding genome function, variation and evolution. Here we report the finished sequence and biological annotation of human chromosome 1. Chromosome 1 is gene-dense, with 3,141 genes and 991 pseudogenes, and many coding sequences overlap. Rearrangements and mutations of chromosome 1 are prevalent in cancer and many other diseases. Patterns of sequence variation reveal signals of recent selection in specific genes that may contribute to human fitness, and also in regions where no function is evident. Fine-scale recombination occurs in hotspots of varying intensity along the sequence, and is enriched near genes. These and other studies of human biology and disease encoded within chromosome 1 are made possible with the highly accurate annotated sequence, as part of the completed set of chromosome sequences that comprise the reference human genome.

The finished sequence of human chromosome 10 comprises a total of 131,666,441 base pairs. It represents 99.4% of the euchromatic DNA and includes one megabase of heterochromatic sequence within the pericentromeric region of the short and long arm of the chromosome. Sequence annotation revealed 1,357 genes, of which 816 are protein coding, and 430 are pseudogenes. We observed widespread occurrence of overlapping coding genes (either strand) and identified 67 antisense transcripts. Our analysis suggests that both inter- and intrachromosomal segmental duplications have impacted on the gene count on chromosome 10. Multispecies comparative analysis indicated that we can readily annotate the protein-coding genes with current resources. We estimate that over 95% of all coding exons were identified in this study. Assessment of single base changes between the human chromosome 10 and chimpanzee sequence revealed nonsense mutations in only 21 coding genes with respect to the human sequence.

Chromosome 13 is the largest acrocentric human chromosome. It carries genes involved in cancer including the breast cancer type 2 (BRCA2) and retinoblastoma (RB1) genes, is frequently rearranged in B-cell chronic lymphocytic leukaemia, and contains the DAOA locus associated with bipolar disorder and schizophrenia. We describe completion and analysis of 95.5 megabases (Mb) of sequence from chromosome 13, which contains 633 genes and 296 pseudogenes. We estimate that more than 95.4% of the protein-coding genes of this chromosome have been identified, on the basis of comparison with other vertebrate genome sequences. Additionally, 105 putative non-coding RNA genes were found. Chromosome 13 has one of the lowest gene densities (6.5 genes per Mb) among human chromosomes, and contains a central region of 38 Mb where the gene density drops to only 3.1 genes per Mb.

Chromosome 6 is a metacentric chromosome that constitutes about 6% of the human genome. The finished sequence comprises 166,880,988 base pairs, representing the largest chromosome sequenced so far. The entire sequence has been subjected to high-quality manual annotation, resulting in the evidence-supported identification of 1,557 genes and 633 pseudogenes. Here we report that at least 96% of the protein-coding genes have been identified, as assessed by multi-species comparative sequence analysis, and provide evidence for the presence of further, otherwise unsupported exons/genes. Among these are genes directly implicated in cancer, schizophrenia, autoimmunity and many other diseases. Chromosome 6 harbours the largest transfer RNA gene cluster in the genome; we show that this cluster co-localizes with a region of high transcriptional activity. Within the essential immune loci of the major histocompatibility complex, we find HLA-B to be the most polymorphic gene on chromosome 6 and in the human genome.

The myosin heavy chain (MHC) and myosin light chain (MLC) isoforms in skeletal muscle of Rana pipiens have been well characterized. We measured the force-velocity (F-V) properties of single intact fast-twitch fibers from R. pipiens that contained MHC types 1 or 2 (MHC1 or MHC2) or coexpressed MHC1 and MHC2 isoforms. Velocities were measured between two surface markers that spanned most of the fiber length. MHC and MLC isoform content was quantified after mechanics analysis by SDS-PAGE. Maximal shortening velocity (V(max)) and velocity at half-maximal tension (V(P 50)) increased with percentage of MHC1 (%MHC1). Maximal specific tension (P(o)/CSA, where P(o) is isometric tension and CSA is fiber cross-sectional area) and maximal mechanical power (W(max)) also increased with %MHC1. MHC concentration was not significantly correlated with %MHC1, indicating that the influence of %MHC1 on P(o)/CSA and W(max) was due to intrinsic differences between MHC isoforms and not to concentration. The MLC3-to-MLC1 ratio was not significantly correlated with V(max), V(P 50), P(o)/CSA, or W(max). These data demonstrate the powerful relationship between MHC isoforms and F-V properties of the two most common R. pipiens fiber types.

Two open-label, randomized studies determined the bioequivalence of a test preparation (Prefest) of micronized 17 beta-estradiol (E2, CAS 50-28-2) tablets as compared with a reference preparation of micronized E2 tablets in healthy postmenopausal women. In Study 1, 36 fasting subjects received 4 test preparation 0.5-mg E2 tablets in one period and 4 reference preparation 0.5-mg E2 tablets in the other period. In Study 2, 36 fasting subjects received 1 test preparation 2-mg E2 tablet in one period and 1 reference preparation 2-mg E2 tablet in the other period. Blood samples were collected before and after dosing to determine serum concentrations of E2, estrone, and estrone sulfate. The 90% confidence intervals for the ratios of mean Cmax and AUC values (test preparation/reference preparation) for all three analytes were within the prescribed 80%-125% range of bioequivalence. In conclusion, the test preparation 0.5-mg and 2-mg micronized E2 tablets are bioequivalent to the respective strength reference preparation micronized E2 tablets.

The MMPI was administered during the patient-perceived best time of the menstrual cycle and during the patient-perceived worst time of the cycle in order to examine the stability of MMPI profile configurations. Subjects were 214 women who were referred to two metropolitan outpatient premenstrual syndrome (PMS) clinics for moderate to severe premenstrual complaints. This sample was selected from 1,849 intake files after screening by strict selection criteria for PMS. The results indicate that there are wide fluctuations in profile patterns between the best and worst times of the menstrual cycle for a large number of patients. Caution in using the MMPI is strongly advised.