The distribution of ticks of the Ornithodoros moubata complex in different habitats in Malawi, particularly pigsties and houses, was established from a four-phase survey undertaken between 1982 and 1985. The first phase consisted of preliminary interviews, the second phase consisted of a questionnaire to Veterinary Assistants (VAs) in rural areas, the third involved tick collections by VAs and the fourth involved both interviews and tick collections by members of the Central Veterinary Laboratory, Lilongwe. The area in which O. moubata is found in pigsties includes much of the African swine fever (ASF) enzootic area, and it seems likely that the enzootic area could become larger in future. The occurrence of O. moubata in warthog habitats was also investigated.

For the global pig industry, classical (CSF) and African swine fever (ASF) outbreaks are a constantly feared threat. Except for Sardinia, ASF was eradicated in Europe in the late 1990s, which led to a research focus on CSF because this disease continued to be present. However, ASF remerged in eastern Europe in 2007 and the interest in the disease, its control and epidemiology increased tremendously. The similar names and the same susceptible species suggest a similarity of the two viral diseases, a related biological behaviour and, correspondingly, similar epidemiological features. However, there are several essential differences between both diseases, which need to be considered for the design of control or preventive measures. In the present review, we aimed to collate differences and similarities of the two diseases that impact epidemiology and thus the necessary control actions. Our objective was to discuss critically, if and to which extent the current knowledge can be transferred from one disease to the other and where new findings should lead to a critical review of measures relating to the prevention, control and surveillance of ASF and CSF. Another intention was to identify research gaps, which need to be closed to increase the chances of a successful eradication of ASF and therefore for a decrease of the economic threat for pig holdings and the international trade.

BACKGROUND

The recent occurrence and spread of African swine fever (ASF) in Eastern Europe is perceived as a serious risk for the pig industry in the European Union (EU). In order to estimate the potential risk of ASF virus (ASFV) entering the EU, several pathways of introduction were previously assessed separately. The present work aimed to integrate five of these assessments (legal imports of pigs, legal imports of products, illegal imports of products, fomites associated with transport and wild boar movements) into a modular tool that facilitates the visualization and comprehension of the relative risk of ASFV introduction into the EU by each analyzed pathway.

RESULTS

The framework's results indicate that 48% of EU countries are at relatively high risk (risk score 4 or 5 out of 5) for ASFV entry for at least one analyzed pathway. Four of these countries obtained the maximum risk score for one pathway: Bulgaria for legally imported products during the high risk period (HRP); Finland for wild boar; Slovenia and Sweden for legally imported pigs during the HRP. Distribution of risk considerably differed from one pathway to another; for some pathways, the risk was concentrated in a few countries (e.g., transport fomites), whereas other pathways incurred a high risk for 4 or 5 countries (legal pigs, illegal imports and wild boar).

CONCLUSIONS

The modular framework, developed to estimate the risk of ASFV entry into the EU, is available in a public domain, and is a transparent, easy-to-interpret tool that can be updated and adapted if required. The model's results determine the EU countries at higher risk for each ASFV introduction route, and provide a useful basis to develop a global coordinated program to improve ASFV prevention in the EU.

A population of domestic pigs in northern Mozambique with increased resistance to the pathogenic effects of African swine fever (ASF) virus was identified by the high prevalence of circulating antibodies to ASF virus. An attempt was made to establish whether the resistance in this population was heritable. Some of these pigs were acquired and transported to a quarantine facility and allowed to breed naturally. Offspring of the resistant pigs were transferred to a high security facility where they were challenged with two ASF viruses, one of which was isolated from one of the Mozambican pigs and the other a genetically closely-related virus from Madagascar. All but one of the 105 offspring challenged developed acute ASF and died. It therefore appears that the resistance demonstrated by these pigs is not inherited by their offspring, or could not be expressed under the conditions of the experiment. The question remains therefore as to the mechanism whereby pigs in the population from which the experimental pigs were derived co-existed with virulent ASF viruses.

African swine fever virus (ASFV) is the etiological agent of a contagious and often lethal viral disease of domestic pigs. There are no vaccines to control Africa swine fever (ASF). Experimental vaccines have been developed using genetically modified live attenuated ASFVs obtained by specifically deleting virus genes involved in virulence, including the thymidine kinase (TK) gene. TK has been shown to be involved in the virulence of several viruses, including ASFV. Here we report the construction of a recombinant virus (ASFV-G/V-ΔTK) obtained by deleting the TK gene in a virulent strain of ASFV Georgia adapted to replicate in Vero cells (ASFV-G/VP30). ASFV-G/P-ΔTK demonstrated decreased replication both in primary swine macrophage cell cultures and in Vero cells compared with ASFV-G/VP30. In vivo, intramuscular administration of up to 10(6) TCID50 of ASFV-G/V-ΔTK does not result in ASF disease. However, these animals are not protected when challenged with the virulent parental Georgia strain.

African swine fever (ASF) is a notifiable infectious disease, caused by the ASF virus (ASFV), which is a DNA virus belonging to the family Asfarviridae, genus Asfivirus. This disease has gained importance in the last decade after its spread in several countries in Eastern and Central Europe, and more recently, in China. Despite the efforts made to eradicate it, ASF is still present on the Mediterranean island of Sardinia (Italy) and has been since 1978. ASF risk factors on the island have been analysed in previous studies; the role of free-ranging pigs in virus persistence has been suggested, but has not been fully elucidated. The most recent eradication plan provides more stringent measures to combat free-ranging pigs and any kind of illegality in the pig sector. From December 2017 to June 2018, a total of 29 depopulation actions were performed in 13 municipalities in central Sardinia, during which 2,281 free-ranging pigs were culled and more than 50% of them were tested for ASFV and antibody presence (1,218 and 1,416, respectively). A total of 651 pigs were seropositive, with a mean seroprevalence of 53.4% (CI 95% = 50.6-56.3), and 38 were ASFV positive (virus prevalence = 2.6%; CI 95% = 2.1-3.0). To the best of our knowledge, the present study is the first to provide a complete evaluation of this millennial system of pig farming and ASFV prevalence in free-ranging pigs. Furthermore, it has emphasised the necessity of combining the maintenance of an epidemiological surveillance program with continuous education of farmers and other people involved in pig husbandry, based on cultural and economic aspects.

Most Escherichia coli strains in the human intestine are harmless. However, enterohemorrhagic E coli (EHEC) is a foodborne pathogen that causes intestinal disease in humans. Conventionally reared (CONV) mice are inconsistent models for human infections with EHEC because they are often resistant to E coli colonization, in part due to their gastrointestinal (GI) microbiota. Although antibiotic manipulation of the mouse microbiota has been a common means to overcome colonization resistance, these models have limitations. Currently, there are no licensed treatments for clinical EHEC infections and, thus, new tools to study EHEC colonization need to be developed. Here, we used a defined microbiota mouse model, consisting of the altered Schaedler flora (ASF), to characterize intestinal colonization and compare host responses following colonization with EHEC strain 278F2 or non-pathogenic E coli strain MG1655. Significantly higher (P<0.05) levels of both strains were found in feces and cecal and colonic contents of C3H/HeN ASF compared to C3H/HeN CONV mice. GI inflammation was significantly elevated (P<0.05) in the cecum of EHEC 278F2-colonized compared to E. coli MG1655-colonized C3H/HeN ASF mice. In addition, EHEC 278F2 differentially modulated inflammatory-associated genes in colonic tissue of C3H/HeN ASF mice compared to E. coli MG1655-colonized mice. This approach allowed for prolonged colonization of the murine GI tract by pathogenic and non-pathogenic E coli strains, and for evaluation of host inflammatory processes. Overall, this system can be used as a powerful tool for future studies to assess therapeutics, microbe-microbe interactions, and strategies for preventing EHEC infections.

African swine fever (ASF) is one of the most important pig diseases, causing high case fatality rate and trade restrictions upon reported outbreaks. In Uganda, a low-income country with the largest pig population in East Africa, ASF is endemic. Animal disease impact is multidimensional and include social and economic impact along the value chain. In low-income settings, this impact keep people poor and push those that have managed to escape poverty back again. If the diseases can be controlled, their negative consequences can be mitigated. However, to successfully argue for investment in disease control, its cost-benefits need to be demonstrated. One part in the cost-benefit equations is disease impact quantification. The objective of this study was therefore to investigate the socio-economic impact of ASF outbreaks at household level in northern Uganda. In a longitudinal study, structured interviews with two hundred, randomly selected, pig-keeping households were undertaken three times with a six month interval. Questions related to family and pig herd demographics, pig trade and pig business. Associations between ASF outbreaks and economic and social impact variables were evaluated using linear regression models. The study showed that pigs were kept in extreme low-input-low-output farming systems involving only small monetary investments. Yearly incidence of ASF on household level was 19%. Increasing herd size was positively associated with higher economic output. The interaction between ASF outbreaks and the herd size showed that ASF outbreaks were negatively associated with economic output at the second interview occasion and with one out of two economic impact variables at the third interview occasion. No significant associations between the social impact variables included in the study and ASF outbreaks could be established. Trade and consumption of sick and dead pigs were coping strategies used to minimize losses of capital and animal protein. The results indicate that causality of social and economic impact of ASF outbreaks in smallholder systems is complex. Pigs are mostly kept as passive investments rather than active working capital, complicating economic analyses and further disqualifying disease control arguments based only on standard economic models.

OBJECTIVE

The purpose of this study was to estimate the volume of normal pancreas in adults using the CT volumetry (summation of the areas technique), analyze the correlation between the volume and the diameters of pancreas, which are measurable by the cross-sectional imaging, and assess the relationship with the gender, age, and body constitution.

METHODS

220 CT examinations were analyzed retrospectively (102 females, 118 males; age 16-82, average 56). Following diameters were measured: cranial-caudal-CC(pancreas), CC(body&tail), CC(body), CC(head); anterior-posterior--AP(tail), AP(body), AP(head); lengths--LL(head), L(body&tail); and maximal transversal diameter of the L1 vertebral body (LL(L1)) and thickness of the abdominal subcutaneous fat (AP(ASF)), as markers of body constitution.

RESULTS

The average volume of the pancreas was 79.2 ± 24.1 cm3 (ranging from 37.4 to 168.2 cm3). Pancreatic volume strongly correlated with all measured diameters of the pancreas (P < 0.0001). Pancreatic volume significantly correlated with gender (M:F = 86.1:72.8 cm3, P = 0.002) and the LL(L1) (r = 0.185, P = 0.008), and did not correlate with the age (r = -0.110, P = 0.151) and the AP(ASF) (r = -0.115, P = 0.104). Correlation of vertebral body-pancreas volume ratio of each subject and the age was strongly negative (r = -0.202, P = 0.006).

CONCLUSIONS

Marked individual variations in normal pancreas volume were observed. Pancreatic volume could be computed using the diameters measurable by the cross-sectional imaging employing the formula: V = (AP(tail) + AP(body))/2 × L(body&tail) × CC(body) + (AP(head)/2)2 × 3.14 × CC(head).

Convalescent clinically normal pigs were tested for the persistence of African swine fever (ASF) infection. One group of pigs was examined 135 days after inoculation with a Brazilian isolate and a 2nd group was examined 110 days after inoculation with a Dominican Republic isolate. Susceptible pigs exposed by contact to these groups remained clinically normal. These contact pigs plus 2 more pigs added to each group developed ASF after being fed and inoculated with tissues collected from recovered pigs. African swine fever virus was not isolated in swine buffy coat cultures inoculated with supernatant fluid from the collected tissues. The remaining convalescent Brazilian and Dominican Republic ASF pigs were challenge inoculated with homologous virus and then with Lisbon 60 ASF virus. Pigs in both groups remained clinically normal after homologous virus challenge inoculation. Pigs in the Brazilian group remained clinically normal after inoculation of the lisbon 60 ASF viral isolate. Of 5 pigs in the Dominican Republic group, 3 developed a transient viremia after inoculation of the Lisbon 60 ASF viral isolate.

Since there is no vaccine available, prevention, control, and eradication of African swine fever (ASF) is based on the implementation of appropriated surveillance and strict sanitary measures. Success of surveillance activities depends on the availability of the most appropriate diagnostic tests. Although a number of good validated ASF diagnostic techniques are available, the interpretation of the ASF diagnostic results can be complex. The reasons lie in the complexity of the epidemiology with different scenarios, as well as in the characteristics of the viruses circulating giving rise to a wide range of clinical forms of ASF. This review provides guidance for an accurate interpretation of ASF diagnostic results linked to the different clinical presentations ranging from per-acute to chronic disease, including apparently asymptomatic infections.

African swine fever (ASF), classical swine fever (CSF), and foot-and-mouth disease (FMD) are highly contagious animal diseases of significant economic importance. Pigs infected with ASF and CSF viruses (ASFV and CSFV) develop clinical signs that may be indistinguishable from other diseases. Likewise, various causes of vesicular disease can mimic clinical signs caused by the FMD virus (FMDV). Early detection is critical to limiting the impact and spread of these disease outbreaks, and the ability to perform herd-level surveillance for all 3 diseases rapidly and cost effectively using a single diagnostic sample and test is highly desirable. This study assessed the feasibility of simultaneous ASFV, CSFV, and FMDV detection by multiplex reverse transcription real-time polymerase chain reaction (mRT-qPCR) in swine oral fluids collected through the use of chewing ropes. Animal groups were experimentally infected independently with each virus, observed for clinical signs, and oral fluids collected and tested throughout the course of infection. All animal groups chewed on the ropes readily before and after onset of clinical signs and before onset of lameness or serious clinical signs. ASFV was detected as early as 3 days postinoculation (dpi), 2-3 days before onset of clinical disease; CSFV was detected at 5 dpi, coincident with onset of clinical disease; and FMDV was detected as early as 1 dpi, 1 day before the onset of clinical disease. Equivalent results were observed in 4 independent studies and demonstrate the feasibility of oral fluids and mRT-qPCR for surveillance of ASF, CSF, and FMD in swine populations.

Cholangiocyte senescence has been linked to primary sclerosing cholangitis (PSC). Persistent secretion of growth factors by senescent cholangiocytes leads to the activation of stromal fibroblasts (ASFs), which are drivers of fibrosis. The activated phenotype of ASFs is characterized by an increased sensitivity to apoptotic stimuli. Here, we examined the mechanisms of apoptotic priming in ASFs and explored a combined targeting strategy to deplete senescent cholangiocytes and ASFs from fibrotic tissue to ameliorate liver fibrosis. Using a coculture system, we determined that senescent cholangiocytes promoted quiescent mesenchymal cell activation in a platelet-derived growth factor (PDGF)-dependent manner. We also identified B-cell lymphoma-extra large (Bcl-xL) as a key survival factor in PDGF-activated human and mouse fibroblasts. Bcl-xL was also up-regulated in senescent cholangiocytes. In vitro, inhibition of Bcl-xL by the small molecule Bcl-2 homology domain 3 mimetic, A-1331852, or Bcl-xL-specific small interfering RNA induced apoptosis in PDGF-activated fibroblasts, but not in quiescent fibroblasts. Likewise, inhibition of Bcl-xL reduced the survival and increased apoptosis of senescent cholangiocytes, compared to nonsenescent cells. Treatment of multidrug resistance 2 gene knockout (Mdr2-/- ) mice with A-1331852 resulted in an 80% decrease in senescent cholangiocytes, a reduction of fibrosis-inducing growth factors and cytokines, decrease of α-smooth muscle actin-positive ASFs, and finally in a significant reduction of liver fibrosis.

Bcl-xL is a key survival factor in ASFs as well as in senescent cholangiocytes. Treatment with the Bcl-xL-specific inhibitor, A-1331852, reduces liver fibrosis, possibly by a dual effect on activated fibroblasts and senescent cholangiocytes. This mechanism represents an attractive therapeutic strategy in biliary fibrosis. (Hepatology 2018;67:247-259).

Facilitation of the spinal monosynaptic reflex by auditory stimulation has been demonstrated previously in animals and man. Analysis of the time course of audiospinal facilitation (ASF) in normal subjects is reported. The role of the cerebral cortex in the control of audiospinal facilitation was investigated in 32 patients with anatomically well-circumscribed lesions, the precise topography of which was determined stereotaxically. Lesions of the caudal part (Heschl's gyrus and temporal plane) of the superior temporal gyrus selectively depressed ASF evoked by contralateral auditory stimulation. In contrast, lesions in temporal, parietal and occipital lobes had no effect. Results obtained with frontal lobe lesions were not homogeneous. The specific involvement of auditory cortex in the gating of behavioral audiomotor reactions is discussed.

For proper attenuation correction of SPECT images, a set of 3D attenuation maps specific to the imaging slices is needed. Among the many different approaches for deriving the attenuation maps, fan beam transmission CT (FBTCT), performed on the same SPECT system as emission imaging, has many promising and clinically practical features. The major problem of FBTCT is that the current SPECT systems do not have a large enough field of view (FOV) to cover the typical cross-sectional size of patients. To address this problem, we have developed a novel asymmetric fan (AsF) sampling scheme to extend the FOV to practical sizes for clinical TCT imaging on existing SPECT systems. This AsF scheme samples only half of the intended FOV in each projection; the other half would be sampled in an opposing projection after detector rotation. We have implemented the AsF sampling on a three-head SPECT system through a specially designed source-collimator assembly. We have modified the conventional convolution backprojection algorithm to facilitate simple and fast image reconstruction. The feasibility of the approach is confirmed by the quality of the derived TCT images of various phantoms and human subjects. The AsF sampling scheme could also have applications in other general transmission CT systems.

OBJECTIVE

The aim of this study was to determine the skeletal effects of Butea total extract (BTE) and its acetone soluble fraction (ASF) from Butea monosperma, which is rich in methoxyisoflavones, in ovariectomized (OVx) rats, a model for postmenopausal bone loss.

METHODS

BTE (1.0 g kg d) and ASF (100 mg kg d) were given orally for 12 weeks to adult OVx rats. The sham-operated and ovariectomy + vehicle groups served as controls. Bone mineral density, osteoid formation (mineral apposition rate and bone formation rate), bone microarchitecture, and bone turnover/resorption markers were studied. Phytoestrogens in rats given BTE and ASF were analyzed by high-performance liquid chromatography. One-way analysis of variance was used to test significance of effects.

RESULTS

OVx rats treated with either BTE or ASF exhibited increased bone mineral density in trabecular bones and improved trabecular microarchitecture compared with the ovariectomy + vehicle group. ASF treatment was more efficient than BTE treatment in maintaining trabecular microarchitecture. Serum osteocalcin and urinary type 1 collagen levels in OVx rats treated with either BTE or ASF were significantly lower than those of the ovariectomy + vehicle group. ASF treatment led to increased mineral apposition rate and bone formation rate compared with ovariectomy + vehicle, whereas BTE had no such effect. In the uterotropic assay, BTE was mildly estrogenic in adult OVx rats. In immature rats, BTE exhibited both estrogenicity and antiestrogenicity. ASF had neither uterine estrogenicity nor antiestrogenicity. Analysis of phytoestrogens revealed significant enrichment of cladrin, isoformononetin, and medicarpin in ASF over BTE.

CONCLUSIONS

Derived from B monosperma, ASF at a 10-fold lower dose than that of BTE was effective in preventing OVx-induced bone loss and stimulated new-bone formation.

Several rifamycin derivatives inhibited the DNA-dependent RNA polymerase of African swine fever (ASF) virus particles. The inhibition was similar to that found with vaccinia virus RNA polymerase. Coumermycin A1, an inhibitor of type II DNA topoisomerases, inhibited strongly RNA synthesis in vitro by ASF virus particles. This suggests that transcription of ASF virus DNA requires a DNA topoisomerase.

African swine fever (ASF) is the most important disease that constrains pig production in Mozambique. Until 1994 it was apparently restricted to the central and northern provinces, but since 1994 outbreaks have been experienced throughout the country. ASF causes severe economic losses both in the small commercial sector and among the large numbers of small-scale producers in the family sector in rural and peri-urban areas. The history of ASF in Mozambique since its first confirmation in 1960 is briefly reviewed, recent outbreaks are reported, and the available information on the virus genotypes that have been responsible for some of the outbreaks is presented. Epidemiological factors that contribute to ASF outbreaks and strategies for limiting the negative effects of the disease in the different pig farming sectors in Mozambique, including raising farmer and community awareness, are discussed.

This paper reports apoptosis of lymph-node lymphocytes in swine experimentally inoculated with a virulent African swine fever (ASF) virus isolate (Malawi '83). Apoptosis was observed in both compartments of cortical tissue, but was more intense in diffuse lymphoid tissue (T area). Lymphopenia detected in peripheral blood was associated with T-lymphocyte depletion. No evidence of ASF virus replication was observed in lymphocytes in the lymph nodes studied. This finding, together with the high rate of virus replication recorded in macrophages in diffuse lymphoid tissue as compared with the low rate recorded for lymphoid follicles, suggests a mechanism for the induction of apoptosis related to virus replication in cells of the mononuclear phagocyte system.

Inflammatory processes and the mechanisms by which they are initiated and controlled within the central nervous system (CNS) involve a vast array of cell types and molecules. One cell type believed to be involved in the control of inflammation in the CNS is the microglial cell. The TLD antibodies are a panel of monoclonal antibodies reactive to rat microglial antigens. One antibody from this panel, clone TLD-1A8A, is specific for antisecretory factor (ASF). ASF is a previously identified protein characterized as a potent inhibitor of enterotoxin-induced intestinal fluid secretion. Our results extend the function of this molecule to include the regulation of immune reactions. Administration of TLD-1A8A to T-cell proliferation or mixed leukocyte response assays resulted in increased proliferation of T cells. Flow cytometric analysis indicates that ASF is expressed on macrophages, B cells and dendritic cells, but not on T cells or granulocytes. Immunohistochemical analysis indicates that ASF is expressed by macrophages and cells of dendritic morphology in the spleen, thymus, lymph nodes, Peyer's patch, and in the perivascular area in the CNS. Furthermore, Western blot analysis indicates that ASF is expressed in many tissues including all secondary lymphoid organs. These data suggest that ASF may have a previously unsuspected role in regulating the immune system.

This paper reviews the literature on cultural and household level constraints on the consumption of animal source foods (ASF). Food proscriptions do not appear to significantly limit food consumption in the areas where this question has been examined, with the possible exception of adult women in some settings. Preferential food allocation patterns, based on economic contribution, social valuation and other factors do play a role in limiting the intake of animal source foods for children and women in some settings. Economic factors limit access to ASF at the household level. Child care patterns in different cultural settings can also serve to enhance or limit the consumption of ASF. Social marketing and positive deviance approaches have had some success in increasing the intake of ASF by using formative research and participatory approaches. The paper concludes with a series of research questions for further investigation.

OBJECTIVE

To analyze the effects of projection-view (PV) distribution on the contrast and spatial blurring of microcalcifications on the tomosynthesized slices (X-Y plane) and along the depth (Z) direction for the same radiation dose in digital breast tomosynthesis (DBT).

METHODS

A GE GEN2 prototype DBT system was used for acquisition of DBT scans. The system acquires PV images from 21 angles in 3° increments over a ±30° range. From these acquired PV images, the authors selected six subsets of PV images to simulate DBT of different angular ranges and angular increments. The number of PV images in each subset was fixed at 11 to simulate a constant total dose. These different PV distributions were subjectively divided into three categories: uniform group, nonuniform central group, and nonuniform extreme group with different angular ranges and angular increments. The simultaneous algebraic reconstruction technique (SART) was applied to each subset to reconstruct the DBT slices. A selective diffusion regularization method was employed to suppress noise. The image quality of microcalcifications in the reconstructed DBTs with different PV distributions was compared using the DBT scans of an American College of Radiology phantom and three human subjects. The contrast-to-noise ratio (CNR) and the full width at half maximum (FWHM) of the line profiles of microcalcifications within their in-focus DBT slices (parallel to detector plane) and the FWHMs of the interplane artifact spread function (ASF) in the Z-direction (perpendicular to detector plane) were used as image quality measures.

RESULTS

The results indicate that DBT acquired with a large angular range or, for an equal angular range,with a large fraction of PVs at large angles yielded superior ASF with smaller FWHM in the Z-direction. PV distributions with a narrow angular range or a large fraction of PVs at small angles had stronger interplane artifacts. In the X-Y focal planes, the effect of PV distributions on spatial blurring depended on the directions. In the X-direction (perpendicular to the chestwall), the normalized line profiles of the calcifications reconstructed with the different PV distributions were similar in terms of FWHM; the differences in the FWHMs between the different PV distributions were less than half a pixel. In the Y-direction (x-ray source motion), the normalized line profiles of the calcifications reconstructed with PVs acquired with a narrow angular range or a large fraction of PVs at small angles had smaller FWHMs and thus less blurring of the line profiles. In addition, PV distributions with a narrow angular range or a large fraction of PVs at small angles yielded slightly higher CNR than those with a wide angular range for small, subtle microcalcifications; however, PV distributions had no obvious effect on CNR for relatively large microcalcifications.

CONCLUSIONS

PV distributions affect the image quality of DBT. The relative importance of the impact depends on the characteristics of the signal and the direction (perpendicular or parallel) relative to the direction of x-ray source motion. For a given number of PVs, the angular range and the distribution of the PVs affect the degree of in-plane and interplane blurring in opposite ways. The design of the scan parameters of tomosynthesis systems would require proper consideration of the characteristics of the signals of interest and the potential trade-off of the image quality of different types of signals.

A missense mutation at codon 640 in the APC gene was identified in a familial adenomatous polyposis (FAP) patient, however, its pathological consequence remained unclear. Here we found that this missense mutation interferes at the nucleotide level with an exonic splicing regulatory element and leads to aberrant splicing of the mutant APC transcript rather than exerting its effect through the observed amino acid change. Analysis of the patient RNA revealed complete skipping of exon 14 in transcripts from the mutant APC allele, leading to a frameshift and a premature stop codon. When cloned into a splicing reporter minigene and transfected into colorectal cell lines, the exon 14 point mutation c.1918C>G (pR640G) was found sufficient to cause the observed exon skipping. Bioinformatic analysis predicted the mutation to change SRp55, hnRNP A1 or ASF/SF2 splicing factor binding sites. Using RNA interference methodology these predictions were experimentally validated and revealed that only ASF/SF2 was required for exon 14 inclusion. These research data identify APC mutation c.1918C>G (pR640G) as pathogenic and indicate a mechanism involving disruption of an ASF/SF2 exonic splicing enhancer element. The results allow genetic diagnosis of a hereditary tumour predisposition but also illustrate the need to complement in silico prediction by splicing reporter assays.