Development and validation of the African Women Awareness of CANcer (AWACAN) tool for breast and cervical cancer.

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.

Pedigree analysis of 5 swine breeds in the United States and the implications for genetic conservation.

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.

Genotype by environment interaction for production traits while accounting for heteroscedasticity.

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.

Student perceptions of tutor skills in problem-based learning tutorials.

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.

Smart training: the US Army's pilot project for combat trauma surgical training.

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.

Antimalarial antisense activity of hexitol nucleic acids.

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.

Molecular cloning and characterization of the Plasmodium falciparum cytidine triphosphate synthetase gene.

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.

Cloning and expression of a prokaryotic enzyme, arginine deiminase, from a primitive eukaryote Giardia intestinalis.

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.

Inhibition of Plasmodium falciparum proliferation in vitro by ribozymes.

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.

Long-term cultivation of Plasmodium falciparum in media with commercial non-serum supplements.

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.

Nucleotide variation in the cytidine triphosphate synthetase gene of Giardia duodenalis.

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.

Isolation, characterization and expression of the gene encoding cytidine triphosphate synthetase from Giardia intestinalis.

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.

Optimal contemporary group structure to maximize genetic progress through genetic evaluation of swine.

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.

A cytidine triphosphate synthetase gene in Plasmodium falciparum.

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.

Subfamilies of serine tRNA genes in the bovine genome.

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.

Efficacy of alternative multivariate best linear unbiased prediction models for genetic evaluation of swine.

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.

A simple method for the purification of mitochondrial DNA from Plasmodium falciparum.

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.

Genetic improvement programs in livestock: swine testing and genetic evaluation system (stages).

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.