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Purposes: The aim of our study is to determine the frequency of vaccine hesitancy and the associated factors among parents with preschool children in a city center., Design and Methods: We conducted a cross-sectional study among parents of preschool children in the central district of Edirne. The cluster sampling method was applied. The data collection tools consisted of a questionnaire designed by the researchers and The Parent Attitudes about Childhood Vaccines (PACV) Scale. The correlations between vaccine hesitancy among parents factos such as the vaccine's content, adverse effects, religious beliefs, media coverage, reliance on community leaders, and the usage of complementary and alternative medical methods were examined., Results: The study group consisted of 315 individuals and the prevalence of vaccine hesitancy among participants was determined to be 13%. The mean PACV scale score of the parents was 29.76 ± 16.62. According to multivariate analyses, the odds ratios (%95 CI) of variables that were statistically significantly associated with parental vaccine hesitancy were as follows: being a mother 8.77 (1.02-75.07), vaccine side effects 2.91 (0.93-9.16), low/equal income level 6.80 (2.45-18.86), low/high income level 5.74 (1.65-20.0), religious reasons 3.33 (1.34-8.25), and media influence 9.53 (3.75-24.18)., Conclusion: Overall, vaccine hesitancy was observed at one in every 7 parents in the central district of Edirne province, and the main determinants of hesitancy were low-income status, media influence and religious beliefs, advers effects and being mother. Community-based intervention programs should be established to combat vaccine hesitancy., Practical Implications: The PACV scale is recommended for detecting community-based vaccine hesitancy., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have potentially influenced the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

BACKGROUND An integrated nonprofit health care system with 13 North Carolina medical centers conducted a time-pressured quality improvement simulation of its plan to implement the "North Carolina Protocol for Allocating Scarce Inpatient Critical Care Resources in a Pandemic" attendant to pandemic scenario planning. Simulation objectives included assessing the plan in terms of a) efficiency and effectiveness; b) comorbidity scoring validity; c) impact by race/ethnicity, gender, age, and payer status; and d) simulation participant impressions of potential impact on clinicians. METHOD The simulation scenario involved scoring 14 patients with the constraint that only 10 could be afforded critical care resources. Also included were independent scoring validation by four clinicians, structured debriefs with simulation participants and observers, and tracking patient outcomes for 30 days. RESULTS Triage scoring was identical among four triage teams. Lack of concordance in clinician comorbidity scoring did not alter patient prioritization for withdrawal of treatment in this small cohort. Protocol scoring was not correlated with resource utilization or near-term mortality. LIMITATIONS The simulation sample was small and selected when COVID-19 census was temporarily waning. No protocol for pediatric patients was tested. CONCLUSIONS The simulation yielded resource allocation concordance using comorbidity scoring by attending physicians, which significantly accelerated triage team decision-making and did not result in notable disparities by race/ethnicity, gender, or advanced age. Qualitative findings surfaced tensions in balancing de-identified data with individualized assessment and in trusting the clinical judgments of other physicians. Additional research is needed to validate the protocol's predictive value related to patient outcomes., (©2021 by the North Carolina Institute of Medicine and The Duke Endowment. All rights reserved.)

Malignancies are often associated with autoimmune diseases, which are addressed by treating the underlying cancer. However, there are rare malignancies that can cause autoimmune diseases even after appropriate treatment. Our patient is a 39-year-old Hispanic man with a malignant thymoma recently treated with chemotherapy and radiation who presented with syncope and dyspnoea. He was found to be both anaemic and thrombocytopenic. His labs were consistent with autoimmune haemolytic anaemia (AIHA), except his reticulocyte count was unexpectedly low. Bone marrow biopsy supported a diagnosis of Evans syndrome, a rare autoimmune condition characterised by (AIHA) combined with immune thrombocytopenia. He was also found to have an acute parvovirus B19 infection. He was treated with steroids and RBC transfusion. His blood counts gradually returned to baseline, with improvement in symptoms. This patient's thymoma treatment and active parvovirus B19 infection likely both played a role in the development of Evans syndrome., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2020. No commercial re-use. See rights and permissions. Published by BMJ.)

Background: Measuring factors influencing time to presentation is important in developing and evaluating interventions to promote timely cancer diagnosis, yet there is a lack of validated, culturally relevant measurement tools. This study aimed to develop and validate the African Women Awareness of CANcer (AWACAN) tool to measure awareness of breast and cervical cancer in Sub-Saharan Africa (SSA)., Methods: Development of the AWACAN tool followed 4 steps: 1) Item generation based on existing measures and relevant literature. 2) Refinement of items via assessment of content and face validity using cancer experts' ratings and think aloud interviews with community participants in Uganda and South Africa. 3) Administration of the tool to community participants, university staff and cancer experts for assessment of validity using test-retest reliability (using Intra-Class Correlation (ICC) and adjusted Kappa coefficients), construct validity (comparing expert and community participant responses using t-tests) and internal reliability (using the Kuder-Richarson (KR-20) coefficient). 4) Translation of the final AWACAN tool into isiXhosa and Acholi., Results: ICC scores indicated good test-retest reliability (≥ 0.7) for all breast cancer knowledge domains and cervical cancer risk factor and lay belief domains. Experts had higher knowledge of breast cancer risk factors (p < 0.001), and cervical cancer risk factors (p = 0.003) and symptoms (p = 0.001) than community participants, but similar knowledge of breast cancer symptoms (p = 0.066). Internal reliability for breast cancer risk factors, lay beliefs and symptom and cervical cancer symptom subscales was good with KR-20 values > 0.7, and lower (0.6) for the cervical cancer risk subscale., Conclusion: The final AWACAN tool includes items on socio-demographic details; breast and cervical cancer symptom awareness, risk factor awareness, lay beliefs, anticipated help-seeking behaviour; and barriers to seeking care. The tools showed evidence of content, face, construct and internal validity and test-retrest reliability and are available for use in SSA in three languages., Competing Interests: We have the following interests: This study was funded in part by GlaxoSmithKline (GSK) Africa Non-Communicable Disease Open Lab (via a supporting grant Project Number: 023). GSK provided in-kind scientific and statistical support as part of capacity strengthening. There are no patents, products in development or marketed products to declare. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials.

Background: Because cell-free DNA (cfDNA) analysis facilitates the noninvasive genomic profiling of metastatic castration-resistant prostate cancer (mCRPC), the authors evaluated the association between cfDNA alterations and outcomes and evolution with therapy., Methods: Patients with mCRPC underwent cfDNA genomic profiling using Guardant360, which examines major cancer-associated genes. Clinical factors, therapy information, failure-free survival, and overall survival (OS) were obtained for select patients. The association between genomic alterations and outcomes was investigated., Results: Of 514 men with mCRPC, 482 (94%) had ≥1 circulating tumor DNA (ctDNA) alteration. The most common recurrent somatic mutations were in TP53 (36%), androgen receptor (AR) (22%), adenomatous polyposis coli (APC) (10%), neurofibromin 1 (NF1) (9%), epidermal growth factor receptor (EGFR), catenin beta-1 (CTNNB1), and AT-rich interactive domain-containing protein 1A (ARID1A) (6% each); and BRCA1, BRCA2, and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) (5% each) The most common genes with increased copy numbers were AR (30%), MYC (20%), and BRAF (18%). Clinical outcomes were available for 163 patients, 46 of whom (28.8%) were untreated for mCRPC. A higher number of ctDNA alterations, AR alterations, and amplifications of MYC and BRAF were associated with worse failure-free survival and/or OS. On multivariable analysis, MYC amplification remained significantly associated with OS. Prior therapy and serial profiling demonstrated the evolution of alterations in AR and other genes., Conclusions: ctDNA frequently was detected in this large cohort of "real-world" patients with mCRPC, and the alterations appeared to be similar to previously reported tumor tissue alterations. A higher number of alterations, and AR and MYC alterations, appear to compromise clinical outcomes, suggesting a role for immune checkpoint inhibitors and novel AR and BET inhibitors in selected patients., (© 2019 American Cancer Society.)

Navel injuries caused by friction against the pasture can promote infection, reproductive problems and costly treatments in beef cattle raised in extensive systems. A haplotype-based genome-wide association study (GWAS) was performed for visual scores of navel length at yearling in Nellore cattle ( Bos indicus ) using data from 2,016 animals and 503,088 single nucleotide polymorphism (SNP) markers. The strongest signal ( p = 1.01 × 10 -9 ) was found on chromosome 5 spanning positions 47.9-48.2 Mbp. This region contains introns 3 and 4 and exons 4 and 5 of the high mobility group AT-hook 2 gene ( HMGA2 ). Further inspection of the region with whole genome sequence data of 21 Nellore bulls revealed correlations between counts of the significant haplotype and copy number gains of a ∼6.2 kbp segment of intron 3 of HMGA2 . Analysis of genome sequences from five African B. indicus and four European Bos taurus breeds revealed that the copy number variant (CNV) is indicine-specific. This intronic CNV was then validated through quantitative polymerase chain reaction (qPCR) using Angus animals as copy neutral controls. Importantly, the CNV was not detectable by means of conventional SNP-based GWAS or SNP probe intensity analyses. Given that HMGA2 affects the expression of the insulin-like growth factor 2 gene ( IGF2 ) together with the pleomorphic adenoma gene 1 ( PLAG1 ), and that the latter has been repeatedly shown to be associated with quantitative traits of economic importance in cattle, these findings highlight the emerging role of variants impacting the insulin-like growth factor pathway to cattle breeding.

East Coast fever (ECF) is a fatal sickness affecting cattle populations of eastern, central, and southern Africa. The disease is transmitted by the tick Rhipicephalus appendiculatus , and caused by the protozoan Theileria parva parva , which invades host lymphocytes and promotes their clonal expansion. Importantly, indigenous cattle show tolerance to infection in ECF-endemically stable areas. Here, the putative genetic bases underlying ECF-tolerance were investigated using molecular data and epidemiological information from 823 indigenous cattle from Uganda. Vector distribution and host infection risk were estimated over the study area and subsequently tested as triggers of local adaptation by means of landscape genomics analysis. We identified 41 and seven candidate adaptive loci for tick resistance and infection tolerance, respectively. Among the genes associated with the candidate adaptive loci are PRKG1 and SLA 2. PRKG1 was already described as associated with tick resistance in indigenous South African cattle, due to its role into inflammatory response. SLA 2 is part of the regulatory pathways involved into lymphocytes' proliferation. Additionally, local ancestry analysis suggested the zebuine origin of the genomic region candidate for tick resistance.

Background: Cervical cancer is the most prevalent malignancy in sub-Saharan Africa (SSA) with many women only seeking professional help when they are experiencing symptoms, implying late-stage malignancy and higher mortality rates. This ecological study assesses population-level exposures of SSA women to the numerous risk factors for HPV infection and cervical cancer, against late-stage presentation of cervical cancer., Materials and Method: A literature review revealed the relevant risk factors in SSA. Open-access databases were mined for variables closely representing each risk factor. A proxy for late-stage presentation was used (ratio of incidence-to-mortality, IMR), and gathered from IARC's GLOBOCAN 2012 database. Variables showing significant correlation to the IMR were used in stepwise multiple regression to quantify their effect on the IMR., Results: Countries with high cervical cancer mortality rates relative to their incidence have an IMR nearer one, suggesting a larger proportion of late-stage presentation. Western Africa had the lowest median IMR (1.463), followed by Eastern Africa (IMR = 1.595) and Central Africa (IMR = 1.675), whereas Southern Africa had the highest median IMR (1.761). Variables selected for the final model explain 65.2% of changes seen in the IMR. Significant predictors of IMR were GDP (coefficient = 2.189 × 10 -6 , p = 0.064), HIV infection (-1.936 × 10 -3 , p = 0.095), not using a condom (-1.347 × 10 -3 , p = 0.013), high parity (-1.744 × 10 -2 , p = 0.008), and no formal education (-1.311 × 10 -3 , p < 0.001)., Conclusion: Using an IMR enables identification of factors predicting late-stage cervical cancer in SSA including: GDP, HIV infection, not using a condom, high parity and no formal education., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Aim: We studied externally controlled anticancer effects of binding tumor growth inhibiting synthetic peptides to magnetoelectric nanoparticles (MENs) on treatment of glioblastomas., Methods: Hydrothermally synthesized 30-nm MENs had the core-shell composition of CoFe 2 O 4 @BaTiO 3 . Molecules of growth hormone-releasing hormone antagonist of the MIA class (MIA690) were chemically bound to MENs. In vitro experiments utilized human glioblastoma cells (U-87MG) and human brain microvascular endothelial cells., Results: The studies demonstrated externally controlled high-efficacy binding of MIA690 to MENs, targeted specificity to glioblastoma cells and on-demand release of the peptide by application of d.c. and a.c. magnetic fields, respectively., Conclusion: The results support the use of MENs as an effective drug delivery carrier for growth hormone-releasing hormone antagonists in the treatment of human glioblastomas.

The recent evolution of cattle is marked by fluctuations in body size. Height in the Bos taurus lineage was reduced by a factor of ~1.5 from the Neolithic to the Middle Ages, and increased again only during the Early Modern Ages. Using haplotype analysis, we found evidence that the bovine PLAG1 mutation (Q) with major effects on body size, weight and reproduction is a >1,000 years old derived allele that increased rapidly in frequency in Northwestern European B. taurus between the 16 th and 18 th centuries. Towards the 19 th and 20 th centuries, Q was introgressed into non-European B. taurus and Bos indicus breeds. These data implicate a major role of Q in recent changes in body size in modern cattle, and represent one of the first examples of a genomic sweep in livestock that was driven by selection on a complex trait.

Superoxide is the proximal reactive oxygen species (ROS) produced by the mitochondrial respiratory chain and plays a major role in pathological oxidative stress and redox signaling. While there are tools to detect or decrease mitochondrial superoxide, none can rapidly and specifically increase superoxide production within the mitochondrial matrix. This lack impedes progress, making it challenging to assess accurately the roles of mitochondrial superoxide in cells and in vivo. To address this unmet need, we synthesized and characterized a mitochondria-targeted redox cycler, MitoParaquat (MitoPQ) that comprises a triphenylphosphonium lipophilic cation conjugated to the redox cycler paraquat. MitoPQ accumulates selectively in the mitochondrial matrix driven by the membrane potential. Within the matrix, MitoPQ produces superoxide by redox cycling at the flavin site of complex I, selectively increasing superoxide production within mitochondria. MitoPQ increased mitochondrial superoxide in isolated mitochondria and cells in culture ~a thousand-fold more effectively than untargeted paraquat. MitoPQ was also more toxic than paraquat in the isolated perfused heart and in Drosophila in vivo. MitoPQ enables the selective generation of superoxide within mitochondria and is a useful tool to investigate the many roles of mitochondrial superoxide in pathology and redox signaling in cells and in vivo., (Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Peranakan Ongole (PO) is a major Indonesian Bos indicus breed that derives from animals imported from India in the late 19(th) century. Early imports were followed by hybridization with the Bos javanicus subspecies of cattle. Here, we used genomic data to partition the ancestry components of PO cattle and map loci implicated in birth weight., Results: We found that B. javanicus contributes about 6-7% to the average breed composition of PO cattle. Only two nearly fixed B. javanicus haplotypes were identified, suggesting that most of the B. javanicus variants are segregating under drift or by the action of balancing selection. The zebu component of the PO genome was estimated to derive from at least two distinct ancestral pools. Additionally, well-known loci underlying body size in other beef cattle breeds, such as the PLAG1 region on chromosome 14, were found to also affect birth weight in PO cattle., Conclusions: This study is the first attempt to characterize PO at the genome level, and contributes evidence of successful, stabilized B. indicus x B. javanicus hybridization. Additionally, previously described loci implicated in body size in worldwide beef cattle breeds also affect birth weight in PO cattle.

Autophagy is a highly conserved degradative process through which cells overcome stressful conditions. Inasmuch as faulty autophagy has been associated with aging, neuronal degeneration disorders, diabetes, and fatty liver, autophagy is regarded as a potential therapeutic target. This review summarizes the present state of knowledge concerning the role of zinc in the regulation of autophagy, the role of autophagy in zinc metabolism, and the potential role of autophagy as a mediator of the protective effects of zinc. Data from in vitro studies consistently support the notion that zinc is critical for early and late autophagy. Studies have shown inhibition of early and late autophagy in cells cultured in medium treated with zinc chelators. Conversely, excess zinc added to the medium has shown to potentiate the stimulation of autophagy by tamoxifen, H2O2, ethanol and dopamine. The potential role of autophagy in zinc homeostasis has just begun to be investigated. Increasing evidence indicates that autophagy dysregulation causes significant changes in cellular zinc homeostasis. Autophagy may mediate the protective effect of zinc against lipid accumulation, apoptosis and inflammation by promoting degradation of lipid droplets, inflammasomes, p62/SQSTM1 and damaged mitochondria. Studies with humans and animal models are necessary to determine whether autophagy is influenced by zinc intake.

Objective: Omega-3 polyunsaturated fatty acid (ω-3 PUFA) supplementation during pregnancy remains controversial. We sought to examine the effects of ω-3 PUFA on inflammation and oxidative stress in vitro and in vivo using a model of preterm labor., Methods: In vivo. Female Swiss Webster mice were fed a normal diet or a 5% fish oil (FO) diet for 3 weeks then mated with normal-fed males. On gestational day 15, dams were injected with either saline (n=10 per group) or lipopolysaccharide (LPS, intrauterine) (n=10 per group). Maternal plasma, amniotic fluid, placentas, and uteri were collected 4 h later and assessed for cytokines; maternal plasma and amniotic fluids were analyzed for oxidative stress. In vitro. RAW264.7 mouse macrophage-like cells were treated with either: vehicle, H2O2, docosahexaenoic acid (DHA), or eicosapentaenoic acid (EPA) (0, 0.1-100 μM) and analyzed for oxidative stress., Results: In vivo. Administration of the 5% FO diet enhanced LPS-induced cytokines in the placenta (P<0.05-0.01) and increased tumor necrosis factor-α in the uterus (P<0.05) and amniotic fluid (P<0.01) when compared to LPS-treated normal-fed animals. Maternal plasma obtained from FO-fed dams showed higher LPS-induced oxidative stress than control-fed animals (P<0.035). However, no differences in oxidative stress were observed in the amniotic fluid. In vitro. Treatment of macrophage-like cells with ω-3 PUFA significantly and dose-dependently increased oxidative stress (P<0.001-0.0001)., Conclusions: Supplementation with FO for prior to and during pregnancy significantly increased LPS-induced inflammation in the amniotic fluid, uterus, and placenta and significantly increased maternal systemic oxidative stress in vivo. Likewise, DHA and EPA induced oxidative stress in macrophage-like cells in vitro.

Background: Globally, cervical cancer is a major public health concern. Cervical cancer is the second most common cancer among women, resulting in approximately 500,000 cases per year. The purpose of this study is to compare disease characteristics between Black Hispanic (BH) and Black non-Hispanic (BNH) women in the US., Materials and Methods: We used stratified random sampling to select cervical cancer patient records from the SEER database (1973-2009). We used Chi-square and independent samples t-test to examine differences in proportions and means., Results: The sample included 2,000 cervical cancer cases of Black non-Hispanic and 91 Black Hispanic women. There were statistically significant differences between black Hispanic and black non- Hispanics in mean age at diagnosis (p<0.001), mean survival time (p<0.001), marital status (p<0.001), primary site of cancer (p<0.001); lymph node involvement (p<0.001); grading and differentiation (p<0.0001); and tumor behavior (p<0.001). Black women were more likely to develop cervical cancer and to have the highest mortality rates from the disease., Conclusions: Findings from this study show clear racial and ethnic disparities in cervical cancer incidence and prognosis that should be addressed.

Background: Race and ethnicity are significant factors in predicting survival time of breast cancer patients. In this study, we applied advanced statistical methods to predict the survival of White non-Hispanic female breast cancer patients, who were diagnosed between the years 1973 and 2009 in the United States (U.S.)., Materials and Methods: Demographic data from the Surveillance Epidemiology and End RESULTS (SEER) database were used for the purpose of this study. Nine states were randomly selected from 12 U.S. cancer registries. A stratified random sampling method was used to select 2,000 female breast cancer patients from these nine states. We compared four types of advanced statistical probability models to identify the best-fit model for the White non- Hispanic female breast cancer survival data. Three model building criterion were used to measure and compare goodness of fit of the models. These include Akaike Information Criteria (AIC), Bayesian Information Criteria (BIC), and Deviance Information Criteria (DIC). In addition, we used a novel Bayesian method and the Markov Chain Monte Carlo technique to determine the posterior density function of the parameters. After evaluating the model parameters, we selected the model having the lowest DIC value. Using this Bayesian method, we derived the predictive survival density for future survival time and its related inferences., Results: The analytical sample of White non-Hispanic women included 2,000 breast cancer cases from the SEER database (1973-2009). The majority of cases were married (55.2%), the mean age of diagnosis was 63.61 years (SD = 14.24) and the mean survival time was 84 months (SD = 35.01). After comparing the four statistical models, results suggested that the exponentiated Weibull model (DIC= 19818.220) was a better fit for White non-Hispanic females' breast cancer survival data. This model predicted the survival times (in months) for White non-Hispanic women after implementation of precise estimates of the model parameters., Conclusions: By using modern model building criteria, we determined that the data best fit the exponentiated Weibull model. We incorporated precise estimates of the parameter into the predictive model and evaluated the survival inference for the White non-Hispanic female population. This method of analysis will assist researchers in making scientific and clinical conclusions when assessing survival time of breast cancer patients.

Background: The use of statistical methods has become an imperative tool in breast cancer survival data analysis. The purpose of this study was to develop the best statistical probability model using the Bayesian method to predict future survival times for the black non-Hispanic female breast cancer patients diagnosed during 1973- 2009 in the U.S., Materials and Methods: We used a stratified random sample of black non-Hispanic female breast cancer patient data from the Surveillance Epidemiology and End RESULTS (SEER) database. Survival analysis was performed using Kaplan-Meier and Cox proportional regression methods. Four advanced types of statistical models, Exponentiated Exponential (EE), Beta Generalized Exponential (BGE), Exponentiated Weibull (EW), and Beta Inverse Weibull (BIW) were utilized for data analysis. The statistical model building criteria, Akaike Information Criteria (AIC), Bayesian Information Criteria (BIC), and Deviance Information Criteria (DIC) were used to measure the goodness of fit tests. Furthermore, we used the Bayesian approach to obtain the predictive survival inferences from the best-fit data based on the exponentiated Weibull model., Results: We identified the highest number of black non-Hispanic female breast cancer patients in Michigan and the lowest in Hawaii. The mean (SD), of age at diagnosis (years) was 58.3 (14.43). The mean (SD), of survival time (months) for black non- Hispanic females was 66.8 (30.20). Non-Hispanic blacks had a significantly increased risk of death compared to Black Hispanics (Hazard ratio: 1.96, 95%CI: 1.51-2.54). Compared to other statistical probability models, we found that the exponentiated Weibull model better fits for the survival times. By making use of the Bayesian method predictive inferences for future survival times were obtained., Conclusions: These findings will be of great significance in determining appropriate treatment plans and health-care cost allocation. Furthermore, the same approach should contribute to build future predictive models for any health related diseases.

Background: Breast cancer is the second leading cause of cancer death for women in the United States. Differences in survival of breast cancer have been noted among racial and ethnic groups, but the reasons for these disparities remain unclear. This study presents the characteristics and the survival curve of two racial and ethnic groups and evaluates the effects of race on survival times by measuring the lifetime data-based half-normal model., Materials and Methods: The distributions among racial and ethnic groups are compared using female breast cancer patients from nine states in the country all taken from the National Cancer Institute's Surveillance, Epidemiology, and End RESULTS cancer registry. The main end points observed are: age at diagnosis, survival time in months, and marital status. The right skewed half-normal statistical probability model is used to show the differences in the survival times between black Hispanic (BH) and black non-Hispanic (BNH) female breast cancer patients. The Kaplan-Meier and Cox proportional hazard ratio are used to estimate and compare the relative risk of death in two minority groups, BH and BNH., Results: A probability random sample method was used to select representative samples from BNH and BH female breast cancer patients, who were diagnosed during the years of 1973-2009 in the United States. The sample contained 1,000 BNH and 298 BH female breast cancer patients. The median age at diagnosis was 57.75 years among BNH and 54.11 years among BH. The results of the half-normal model showed that the survival times formed positive skewed models with higher variability in BNH compared with BH. The Kaplan-Meir estimate was used to plot the survival curves for cancer patients; this test was positively skewed. The Kaplan-Meier and Cox proportional hazard ratio for survival analysis showed that BNH had a significantly longer survival time as compared to BH which is consistent with the results of the half-normal model., Conclusions: The findings with the proposed model strategy will assist in the healthcare field to measure future outcomes for BH and BNH, given their past history and conditions. These findings may provide an enhanced and improved outlook for the diagnosis and treatment of breast cancer patients in the United States.

Objective: To develop a model for the identification of individuals at risk for carotid stenosis (CS) that could be useful in a clinical setting when trying to decide whether screening is worthwhile., Background: Evidence that aggressive medical therapy and life style changes reduce the risk of stroke in individuals with CS is increasing and has led to a renewed interest in screening for CS., Methods: Data on demographics and risk factors were obtained from 2,885,257 individuals who had carotid Duplex scans by Life Line Screening between 2003 and 2008. Multivariable logistic regression analysis was used to identify independent risk factors for CS (>50% stenosis). A scoring system was developed where risk factors were assigned a weighted score. Predictive ability was assessed by calculating C statistics and r2., Results: In the screened cohort, 71,004 patients (2.4%) had CS. Independent risk factors include advanced age, smoking, peripheral arterial disease, high blood pressure, coronary artery disease, diabetes, cholesterol, and abdominal aortic aneurysm. African Americans, Asians, and Hispanics had reduced risk than whites. Exercise and consumption of fruit, vegetables, and nuts had a modest protective effect. A predictive scoring system was created that identifies individuals with CS more efficiently (C statistic = 0.753) than previously published models., Conclusions: We provide a model that enables identification of individuals who have a high probability of having CS. This model can be helpful in designing targeted screening programs that are cost-effective.

As the methodologies available for the detection of positive selection from genomic data vary in terms of assumptions and execution, weak correlations are expected among them. However, if there is any given signal that is consistently supported across different methodologies, it is strong evidence that the locus has been under past selection. In this paper, a straightforward frequentist approach based on the Stouffer Method to combine P-values across different tests for evidence of recent positive selection in common variations, as well as strategies for extracting biological information from the detected signals, were described and applied to high density single nucleotide polymorphism (SNP) data generated from dairy and beef cattle (taurine and indicine). The ancestral Bovinae allele state of over 440,000 SNP is also reported. Using this combination of methods, highly significant (P<3.17×10(-7)) population-specific sweeps pointing out to candidate genes and pathways that may be involved in beef and dairy production were identified. The most significant signal was found in the Cornichon homolog 3 gene (CNIH3) in Brown Swiss (P = 3.82×10(-12)), and may be involved in the regulation of pre-ovulatory luteinizing hormone surge. Other putative pathways under selection are the glucolysis/gluconeogenesis, transcription machinery and chemokine/cytokine activity in Angus; calpain-calpastatin system and ribosome biogenesis in Brown Swiss; and gangliosides deposition in milk fat globules in Gyr. The composite method, combined with the strategies applied to retrieve functional information, may be a useful tool for surveying genome-wide selective sweeps and providing insights in to the source of selection.

There are advantages for use of frozen-thawed boar sperm (FTS) as a tool for preservation and transfer of valuable genetic material, despite its practical limitations. It was hypothesized that increasing the number of motile FTS and number of fixed-time artificial inseminations (AI) would improve pregnancy rate and litter size. Semen from six boars was frozen in 0.5mL straws at 500x10(6)cells/mL. Gilts approximately 170 days of age, were induced into estrus with PG600 and synchronized using MATRIX (synthetic progestagen). Following last feeding of MATRIX (LFM), gilts were checked twice daily for estrus. At onset of estrus, gilts were randomly assigned in a 3x2 factorial treatment design to receive 1x10(9) motile FTS (n=19), 2x10(9) motile FTS (n=19), 4x10(9) motile FTS (n=19) in a single AI at 32h after onset of estrus, or 1x10(9) motile FTS (n=18), 2x10(9) motile FTS (n=17), or 4x10(9) motile FTS (n=19) in each of the two AI at 24 and 32h following onset of estrus. Ultrasonography was performed at 12h intervals after estrus to estimate time of ovulation. Reproductive tracts were collected 28-34 days following AI. Estrus occurred at 139+/-2h (mean+/-SE) after LFM and ovulation at 33+/-1h following onset of estrus. Dose and number of inseminations did not interact or individually influence pregnancy rate at slaughter (73+/-4.2%) or numbers of normal fetuses (10.8+/-0.5). However, number of fetuses tended (P=0.14) to increase with double AI but not with dose. Boar did not affect pregnancy rate but did affect number of normal fetuses and embryonic survival (P<0.01). Longer intervals from insemination to ovulation reduced pregnancy rate (P<0.05), number of normal fetuses (P<0.001), and embryonic survival (P<0.01). Ovarian abnormalities at slaughter were associated with reduced pregnancy rate (P<0.001). The results of this experiment indicate that a double insemination using 2x10(9) motile sperm would produce the greatest number of piglets with fewest numbers of frozen sperm used, while double AI with 1x10(9) motile sperm would be most practical for pig production with limited genetic resources. Fertility was also influenced by boar, interval from insemination to ovulation, and gilt ovarian abnormalities., (2010 Elsevier B.V. All rights reserved.)

Globally, genetic diversity of livestock populations is contracting. Knowing the true extent of the contraction is needed to develop effective conservation strategies. Although contractions of genetic diversity have been documented at the breed level, little within breed documentation has occurred. This situation is no different for US swine breeds. Therefore, the objective of this study was to establish an inbreeding baseline for 5 pig breeds via pedigree records extracted from purebred registrations to each breed association for Berkshire (n = 116,758), Duroc (n = 878,480), Hampshire (n = 744,270), Landrace (n = 126,566), and Yorkshire (n = 727,268). For all breeds the number of registrations peaked after 1990 and declined since that time. The breeder structure was analyzed for Berkshire and Duroc; the average breeder registered pigs for 4.0 yr for both breeds. Breeders were grouped by longevity and herd size, and the inbreeding levels for the current population (pigs born 2006 and later) were evaluated. Presently, more than 99% of all pigs are inbred with the majority having inbreeding less than 10%. The range for percentage of animals that are more than 25% inbred ranged from 1.16% for Yorkshire to 6.09% for Berkshire. The greatest inbreeding for all animals within a breed ranged from 51% for Landrace and 65% for Yorkshire. Sires were grouped into 10 percentiles based on number of great-grandprogeny (GGP) produced; for all breeds, the top 10 percentile accounted for more than 75% of all GGP. Sixty percent of all sires produced less than 1% of all GGP, indicating few males are contributing to future generations. Generations ranged from 17 to 19 per breed with a generation interval ranging from 1.65 yr for Berkshire to 2.21 yr for Yorkshire. Mean inbreeding (%) at generation 17 (the most generations computed across breeds), rate of inbreeding per generation, and effective population size were 12.3, 0.0065, and 77 for Berkshire; 11.8, 0.0044, and 113 for Duroc; 6.8, 0.0046, and 109 for Hampshire; 17.9, 0.0067, and 74 for Landrace; and 8.0, 0.0044, and 113 for Yorkshire, respectively. The 2 breeds with fewest registrations, Berkshire and Landrace, had greater inbreeding rates and smaller effective population sizes, suggesting a need for more immediate conservation efforts. This analysis provides a basis for future monitoring of the genetic diversity of pig breeds and serves as a basis for planning conservation activities.

Grazing (G) provides an alternative management system for dairy production. Heteroscedasticity (HV) of the data may bias estimates of genetic correlations of yield traits between environments, an indicator of genotype-by-environment interaction (GxE). The objective of this study was to investigate the effect of HV on estimates of heritabilities and genetic correlations for mature-equivalent milk, protein, and fat yield, and lactation-average somatic cell scores of daughters, and to determine if HV affects the ability of sire's predicted transmitting ability (PTA) to predict daughter production in G and confinement (C) herds. Data consisted of 72,489 records from 35,674 cows in 366 G herds from 11 states, and 117,629 records from 50,963 cows in 373 C herds from the same 11 states plus 1 geographically contiguous state. Herds were divided into variance quartiles (Q(V)1-Q(V)4) based on milk yield. A transformation was used to reduce HV by standardizing the within-herd standard deviation to the average across-herd standard deviation of a base year for each parity, and was similar to the method used in current USDA-DHIA genetic evaluations. Regression of daughter yield on sire PTA showed that PTA overestimated production of all traits in Q(V)1-Q(V)3 and of milk in Q(V)4 of G herds. For C herds, yields of milk in Q(V)1 and Q(V)2, and of protein and fat in Q(V)1 were overestimated, and protein was underestimated in Q(V)4. Reducing HV had little effect on G herds, but for C herds, regression did not differ from unity for milk and protein in Q(V)1 and Q(V)2. For milk, protein, and fat in G, heritabilities were approximately 0.17, 0.17, and 0.19, respectively. The heritabilities for milk, protein, and fat in C herds were approximately 0.16, 0.17, and 0.21, respectively. Genetic correlations between C and G did not suggest a GxE in 3 upper quartiles, but a possible GxE (correlation = 0.21, estimated standard error = 0.22) for the lowest quartile. Reducing HV did not affect estimates of heritabilities or genetic correlations. Results indicated that modest evidence for existence of GxE did not arise solely from HV.

Purpose: The purpose of this article is to review current evidence available for alpha-lipoic acid (ALA) and its ability to improve symptoms of peripheral diabetic neuropathy (PDN)., Methods: This article searched MEDLINE from 1966 to November 2005 to identify clinical trials that supplemented ALA to individuals with type 1 or type 2 diabetes and positive sensory symptoms of PDN. Clinical trials to be included in this review met specific criteria of randomization, double masking, and placebo-controlled design., Results: The search results produced 5 clinical trials that met the prerequisites for this review. ALA appears to improve neuropathic symptoms and deficits when administered via parenteral supplementation over a 3-week period. Oral treatment with ALA appears to have more conflicting data whether it improves sensory symptoms or just neuropathic deficits alone. An oral regimen of ALA and optimal length of treatment remains unclear. Both parenteral and up to a 2-year time period of oral supplementation of ALA appears to be safe without affecting glycemic control., Conclusions: Based on these results, ALA should be considered as a treatment option for patients with PDN. When discussing supplementation with patients, it is important to discuss potential side effects; vitamin, mineral, and drug interactions; and current evidence available regarding efficacy.

CTP synthetase (E C 6.3.4.2 UTP: ammonia ligase (ADP-forming)) catalyses the formation of CTP from UTP and, in the human parasite Plasmodium falciparum, is the sole source of cytidine nucleotides. It is thus a potential chemotherapeutic target, especially as the gene sequence indicated that the encoded GAT-domain of the enzyme contains two extended peptide segments (42aa and 223aa as compared to the host enzyme). Here, we circumvent the codon usage problems associated with the high A/T content of the P. falciparum sequence, especially evident in sequences encoding the extra peptides, to successfully express active recombinant P. falciparum CTP synthetase using preferred E. coli codons. This partially synthetic gene produced recombinant enzyme, containing the additional segments, which was functionally assayed for activity in vitro. We also show the native enzyme contains the additional peptides using immunoblots with antibodies derived from the recombinant protein. Confocal microscopy, using antibodies to the recombinant protein, provided evidence that the enzyme is expressed in vivo. This establishes for the first time that P. falciparum contain active CTP synthetase and that this enzyme contains two novel insert sequences in the functional enzyme.

BACKGROUND: Hammerhead ribozymes are RNA-based molecules which bind and cleave other RNAs specifically. As such they have potential as laboratory reagents, diagnostics and therapeutics. Despite having been extensively studied for 15 years or so, their wide application is hampered by their instability in biological media, and by the poor translation of cleavage studies on short substrates to long RNA molecules. This work describes a systematic study aimed at addressing these two issues. RESULTS: A series of hammerhead ribozyme derivatives, varying in their hybridising arm length and size of helix II, were tested in vitro for cleavage of RNA derived from the carbamoyl phosphate synthetase II gene of Plasmodium falciparum. Against a 550-nt transcript the most efficient (t1/2 = 26 seconds) was a miniribozyme with helix II reduced to a single G-C base pair and with twelve nucleotides in each hybridising arm. Miniribozymes of this general design were targeted to three further sites, and they demonstrated exceptional cleavage activity. A series of chemically modified derivatives was prepared and examined for cleavage activity and stability in human serum. One derivative showed a 103-fold increase in serum stability and a doubling in cleavage efficiency compared to the unmodified miniribozyme. A second was almost 104-fold more stable and only 7-fold less active than the unmodified parent. CONCLUSION: Hammerhead ribozyme derivatives in which helix II is reduced to a single G-C base pair cleave long RNA substrates very efficiently in vitro. Using commonly available phosphoramidites and reagents, two patterns of nucleotide substitution in this derivative were identified which conferred both good cleavage activity against long RNA targets and good stability in human serum.

Objective: The problem-based learning (PBL) tutor plays a role that is different from the role of a teacher in a conventional teaching format. In the Faculty of Medicine and Health Sciences, United Arab Emirates, all students are Arab nationals and tutors are expatriates with different sociocultural backgrounds from the students. This study was designed to investigate how students evaluate tutors in PBL tutorials and whether student evaluations of tutors change with the progress of students in PBL tutorials., Methods: Differences in tutor performance evaluation by male and female students were also analysed. The students evaluated 12 tutor skills in a scale of 1-3, 1 being 'below average' and 3, 'outstanding'. Student responses from a total of 314 (98.1%) completed forms collected over 2 academic years were analysed statistically. A total of 14 tutors participated in the PBL programme., Results: The analysis revealed that tutors as a group were rated as having average to outstanding tutor skills in 10 items of the evaluation form. Students and faculty perceptions were different for the tutor skills of guiding students for information management. The students expected more support from tutors, whereas the tutors tried to emphasize self-learning in the PBL curriculum. Lower scores to the tutors in the 'problem' bringing sociocultural and religious issues for discussion showed that a gap in sociocultural/religious understanding between students and tutors might influence tutor skills., Conclusions: Differences in tutor evaluation by male and female students indicate necessity of adopting different strategies by tutors in a different sociocultural background. The results of the study have direct implications for faculty development.

Military medical personnel have an important wartime mission. To enhance combat trauma readiness and increase training opportunities, the military is experimenting with providing this additional trauma training in a civilian level 1 trauma center.

Antisense oligonucleotides and ribozymes have shown promise both as antimalarial agents and as tools for identifying genes vital for parasite survival. This task is urgent due to the ineffectiveness of current drug regimes on the most virulent human malarial parasite, Plasmodium falciparum. The development of new ways to modify and/or protect conventional phosphodiester oligonucleotides to improve nuclease resistance is also important. We assessed the effect of antisense oligonucleotides containing phosphorylated anhydrohexitols in suppressing the growth of P. falciparum in culture. The modified oligonucleotides were able to inhibit parasite growth in a sequence-specific manner, but not as well as the phosphorothioated antisense oligonucleotides, which are effective antimalarials at submicromolar concentrations. Two reasons are suggested: the absence of RNase H activation and differences in membrane transport.

Using degenerate oligonucleotides derived from conserved amino acid regions of cytidine triphosphate synthetase, a fragment of the gene from the malarial parasite, Plasmodium falciparum, was isolated by polymerase chain reaction (PCR). This fragment was used as a probe in the isolation of genomic clones containing the entire pfCTP synthetase coding region (2580 bp). The gene encodes the largest CTP synthetase found in any organism to date due to the presence of two additional sequences which are part of the continuous open reading frame and are not introns as their presence in the mRNA was confirmed by reverse transcriptase-PCR. These features distinguish the parasite enzyme from that of the host making it an attractive target for structure based drug design.

Arginine deiminase (EC 3.5.3.6) catalyzes the irreversible catabolism of arginine to citrulline in the arginine dihydrolase pathway. This pathway has been regarded as restricted to prokaryotic organisms but is an important source of energy to the primitive protozoan Giardia intestinalis. In this paper we report the cloning and expression of the arginine deiminase gene from this parasite. Degenerate oligonucleotides based on amino acid sequences of tryptic peptides from the purified protein were used to amplify a portion of the arginine deiminase gene. This was then used as a probe to screen HindIII and PstI "mini" libraries to obtain two overlapping clones that contained the arginine deiminase gene. The open reading frame encoded 581 amino acids including all of the tryptic peptides that were sequenced and corresponded to a molecular mass of 67 kDa. Northern blot analysis identified a single 1.8-kilobase transcript in both trophozoites and encysting cells. Arginine deiminase was successfully expressed in Escherichia coli and purified to homogeneity. The recombinant protein was found to have characteristics comparable with those of the native enzyme.

The cytidine triphosphate synthetase genes from three diverse strains of Giardia duodenalis have been sequenced and found to vary significantly from one another. The isolates were chosen as representatives of three demes as determined by several criteria including divergence in the rDNA repeat unit. Inserts in the genes and protein are conserved in length but are the most divergent regions among the three sequences examined. Variation in the rest of the gene occurs primarily in the third base position resulting in many silent mutations. One of the isolates (1709) was found to contain two genes with high sequence homology.

Catalytic RNA (ribozymes) suppressed the growth of the human malarial parasite Plasmodium falciparum in vitro. The phosphorothioated hammerhead ribozymes targeted unique regions of the P. falciparum carbamoyl-phosphate synthetase II gene. The P. falciparum carbamoyl-phosphate synthetase II gene encodes the first and limiting enzyme in the pathway, and its mRNA transcript contains two large insert regions absent in other carbamoyl-phosphate synthetases, including that from humans. These inserts are ideal targets for nucleic acid therapy. Exogenous delivery of ribozymes to cultures reduced malarial viability up to 55% at 0.5 microM ribozyme concentrations, which is significantly greater than control levels (5-15% reduction), suggesting a sequence-specific inhibition. This inhibition was shown to be stage-specific, with optimal inhibitions being detected after 24 h, coincident with maximal production of the carbamoyl-phosphate synthetase enzyme in the course of the life cycle of the parasite. A decrease in total carbamoyl-phosphate synthetase activity was observed only in cultures treated with the ribozymes. The task of developing alternative therapeutic agents against malaria is urgent due to the evolution of drug-resistant strains of P. falciparum, the most virulent of all human malarial parasites. Another critical issue to be addressed is the possibility of eliminating or reducing any systemic toxicity to the host, which can potentially be provided by nucleic acid therapy. This work is the first reported assessment of the ability of ribozymes as antimalarials. Ribozyme inhibition assays can also aid in identifying important antimalarial loci for chemotherapy. The malarial parasite can, in turn, be a useful in vivo host to study the catalysis and function of new ribozyme designs.

Continuous cultivation of both drug-sensitive and drug-resistant strains of Plasmodium falciparum was achieved using a mixture of commercially available serum substitutes. This provides easier standardisation of experimental data and minimises the risk of infection from handling of human serum. In addition, Nutridoma SR and AlbuMAXI are readily available and are biologically defined. Parasites grown in serum-free medium are morphologically and metabolically identical to those cultivated in medium supplemented with 10% human serum.

The cytidine triphosphate synthetase gene from Giardia intestinalis was cloned using a PCR-based strategy. A 519 bp PCR product was obtained from the amplification of genomic DNA using two oligonucleotides derived from the CTP synthetase amino acid consensus sequences DPYINVDPG and KTKPTQ. This product was used to probe restriction endonuclease digested genomic DNA and the respective plasmid mini-libraries. Two genomic clones were obtained one with a 3.6 kb HindIII DNA fragment, containing approximately three-quarters of the 5'-end of the synthetase gene and subsequently, a 5.8 kb PstI DNA fragment which contained the whole gene. The intronless gene has a 1863 bp open reading frame encoding 620 amino acids (M(r) of 68.3 kDa). A well conserved catalytic glutamine aminotransferase (GAT) domain was identified. In addition, three insert sequences were found which are not present in CTP synthetase from other species. Alignment and comparison of the deduced amino acid sequence relative to CTP synthetases from other species revealed a high degree of identity (34%) with a greater resemblance to prokaryotes than eukaryotes. The gene is located on chromosome 6 and the messenger RNA encoding it is estimated to be 1.9 kb. The coding region of G. intestinalis CTP synthetase was generated by PCR and subsequently cloned into the pQE30 vector for expression in E. coli. This construct yielded a soluble and enzymatically active recombinant protein which was purified by a Ni-NTA affinity column. The purified recombinant protein had a subunit molecular weight of 69.5 kDa and a native molecular weight of approximately 274 kDa. Kinetic studies of the partially purified recombinant G. intestinalis CTP synthetase gave apparent K(m) values of 0.1 mM and approximately 0.5 mM for the substrates UTP and L-glutamine respectively in accord with previously reported values for the native enzyme.

Accuracies of sire EPD were calculated for a typical growth trait and a typical maternal trait for alternative contemporary group structures. For a given family size (number of progeny tested for a sire in a contemporary group), accuracy increased as the number of sires increased and as the number of contemporary groups increased. An exponential equation was found to best predict accuracy from the number of sires and groups. Partial derivatives can be used to determine the optimal number of sires and groups for a given economic situation (fixed number of animals tested per group). It is recommended that progeny of at least two sires be represented in each contemporary group, but having more than five sires does not effectively increase accuracy further. Accuracy increases with a larger herd size, as more sires and more groups of pigs are tested. When the number of litters that can be tested is at its limit, accuracy is maximized with a small number of sires, and progeny divided among several contemporary groups. However, accuracy is only part of the herd's genetic improvement. Selection decisions must be made to avoid inbreeding, which can be a problem, particularly in small herds.

The malarial parasite Plasmodium falciparum is dependent on de novo synthesis for its pyrimidine nucleotide requirements. However, the activity of the key enzyme in cytidine nucleotide synthesis, CTP synthetase (EC 6.3.4.2), has not been reported. We present evidence for a CTP synthetase gene in P. falciparum, having isolated a PCR product obtained using 2 primers derived from the CTP synthetase amino acid consensus sequences DPYINVDPG and GICLGMQ. The amplified DNA segment encodes an amino acid sequence with considerable homology to CTP synthetases from several other species including human and yeast.

A bovine tRNA gene cluster has been characterized and the sequences of four tDNAs determined. Two of the tDNAs could encode tRNA(SerIGA), one tRNA(SerUGA), and the fourth tRNA(GlnCUG). The three serine tDNAs representing the UCN codon isoacceptor family are almost identical. However, the sequence of the tDNA(SerTGA) differs from a previously sequenced bovine tDNA(SerTGA) at 12 positions (ca. 14%). This finding suggests that in the bovine genome, two subfamilies of genes might encode tRNA(SerUGA). It also raises the possibility that new genes for a specific UCN isoacceptor might arise from the genes of a different isoacceptor, and could explain previously observed differences between species in the anticodons of coevolving pairs of tRNAs(SerUCN). The gene cluster also contains complete and partial copies, and fragments, of the BCS (bovine consensus sequence) SINE (short interspersed nuclear element) family, six examples of which were sequenced. Some of these elements occur in close proximity to two of the serine tDNAs.

A comparison of the accuracy of alternative BLUP evaluations of swine performance data is reported. A simulated data set of performance for days to market, backfat, number born alive, number weaned, and litter weight in six herds was used for the evaluation. The data structure was derived by using the performance records of six herds sampled from the American Yorkshire Club's STAGES (Swine Testing and Genetic Evaluation System) database. For growth traits, 10,360 pig records from 129 sires and 897 dams were recorded. For maternal traits, records on 2,598 litters of 1,209 sows from 147 sires and 585 dams were included. The actual observed performance of each record was removed and replaced with simulated performance. These simulated data were analyzed by within- and across-herd BLUP models using STAGES and Animal Model (AM) procedures. Results indicate that the alternative BLUP procedures produced similar estimates. Correlations between STAGES and AM EPD ranged from .84 to .95. Correlations between STAGES EPD and true genetic value (G) ranged from .41 to .74, and correlations between AM EPD and G ranged from .40 to .74. On average, AM EPD had a .04 larger correlation with G than did STAGES EPD, although the difference in the correlations was not significant (P greater than .05). Trends in EPD for sires, dams, and pigs or sows were the same. Likewise, standard errors of prediction for AM EPD averaged 4% smaller than those for STAGES EPD. Computationally, the AM procedures used 15 to 20 times as much processing time as did STAGES procedures.

Genetic evaluations for the U.S. swine industry are conducted by the eight purebred associations of the National Association of Swine Records. Within-herd evaluations of the growth traits (days to 105 kg [market] and backfat depth) were first reported in 1986. Analyses of the maternal traits (litter size at birth and weaning, and litter 21-d weight) were inaugurated in 1987. Expected progeny differences (EPD) are reported for all traits and for general, paternal, and maternal bioeconomic indexes. A sow productivity index combining only maternal traits is available. All records are adjusted according to National Swine Improvement Federation (NSIF) guidelines for effects such as number of pigs transferred at crossfostering and age at recorded observation prior to the BLUP evaluation. Within-herd analyses of individual contemporary groups are conducted immediately on receipt of performance records at each breed association office. All parents in the herd and the young pigs in the current group are evaluated. A report is returned to the breeder for use in herd selection and the EPD are placed in the pedigree file. The genetic base of each herd is defined as the first n tested pigs or litters, where n is the number of pigs registered annually within the herd. Change in mean EPD between groups is indicative of genetic trend. Periodic across-herd analyses are used to update interim within-herd analyses and a national sire summary is published.

Mitochondrial DNA (mtDNA) from Plasmodium falciparum was isolated by conventional differential centrifugation in an SS34 rotor, a simpler method than CsC1 centrifugation of total DNA as employed by other workers. The nature of the sample was verified by sequencing a polymerase chain reaction (PCR) product obtained using oligonucleotide primers derived from known malarial mtDNA sequence.