A minimum 2-year follow-up retrospective review was undertaken to assess our experience with an anterior paramedian muscle-sparing approach to the lumbar spine for anterior spinal fusion (ASF). The records of 28 patients (November 1991 through January 1996) undergoing ASF via a left lower quadrant transverse skin incision (6-10 cm) with a paramedian anterior rectus fascial Z-plasty retroperitoneal approach were reviewed. Diagnosis, number, and level of lumbar interspaces fused, types of fusion, estimated blood loss, length of procedure, length of hospital stay, and complications were analyzed. All cases were completed as either a same-day anterior/posterior (24 of 28) or as a staged procedure at least 1 week after posterior fusion (4 of 28). The General Surgery service performed the muscle-sparing approach, whereas the Orthopedic Spine service performed the ASF. There were 14 men and 14 women, with a mean age of 35.5 years (range, 11-52 years). Diagnoses included spondylolisthesis in 20 cases (including four grade III or IV slips), segmental instability (degenerative or postsurgical) in 7, and 1 flatback deformity. A single level was fused in 20 cases (L4/5 in 4 and L5/S1 in 16), two levels were fused in 5 cases (L4/5 and L5/S1) and three levels were fused in 2 cases (L3/4, L4/5, and L5/S1). The mean length of stay was 7.4 days (range, 5-12 days). The mean estimated blood loss was 300 mL for the anterior procedure alone and 700 ml for both anterior/posterior procedures on the same day. The mean length of operating room time for the anterior approach and fusion was 117 minutes (range, 60-330 minutes). Posterior instrumentation was used in all cases. Anterior interbody struts used included 19 autogenous tricortical grafts, 4 fresh-frozen allografts (2 femoral rings and 2 iliac crests), 3 carbon fiber cages packed with autogenous bone, and a Harms titanium cage with autograft. There was one L5 corpectomy for which a large tricortical allograft strut was utilized. There were no vascular, visceral, or urinary tract injuries. In three cases a mild ileus developed, which resolved spontaneously. We conclude that the anterior paramedian muscle-sparing retroperitoneal approach is safe, uses a small skin incision, avoids cutting abdominal wall musculature, and allows for multiple-level anterior spinal fusions by a variety of interbody fusion techniques. This approach does not require transperitoneal violation or added endoscopic instrumentation, nor does it limit fusion level and technique of fusion, as is the case with the recently popularized laparoscopic approach to the lumbar spine.

A serological survey of free-ranging domestic pigs in the Central and Southern Regions of Malawi, together with laboratory data on confirmed cases of African swine fever (ASF) and data from interviews with pig owners, undertaken over a four-year period from 1981-4, has enabled the ASF enzootic area of Malawi to be identified. The area covers much of the western part of the Central Region and includes Mchinji district and parts of Kasungu, Ntchisi, Dowa and Lilongwe districts. Mortality is substantially less than 100% in outbreaks within the enzootic area but approaches 100% in outbreaks outside this area, as shown by both the serological investigation and the interview data.

In order to re-evaluate functional implications of alphasmooth muscle actin (alphaSMA) expression in lens epithelial cells (LECs), we assessed its presence in donor lenses without visible opacities (DON), lenses with mature cataract (CAT), and cataractous lenses with posterior subcapsular opacities (PSO) or anterior subcapsular fibrosis (ASF). The levels of alphaSMA and transforming growth factor-beta2 (TGFbeta2) mRNAs were measured by classical and real-time PCR. Expression and structural organisation of alphaSMA protein and beta-catenin were monitored by Western blotting and confocal microscopy. All DON analysed contained measurable amounts of alphaSMA mRNA. In CAT without and with PSO, mRNA expression was increased and, again more than doubled in ASF. TGFbeta2 mRNA expression varied widely between the individual samples but was slightly increased in ASF. No correlation existed between alphaSMA or TGFbeta2 expression and the age of the donors in any of the lens categories. Confocal microscopy revealed that, in DON and CAT, alphaSMA was preferentially expressed in a simple granular pattern in single or small clusters of LECs within a normally shaped cobblestone epithelium. Locally, the granules were merged into short stretches at the cell margin. In CAT, a few abnormally shaped cells contained polygonal alphaSMA structures and short stress fibres. In CAT with PSO and ASF, polygons and stress fibre bundles predominated in spindle-shaped cells. Expression patterns of different complexity were often present in the same epithelium. Apical polygons and basal stress fibres were detected within the same cell and may reflect instability of the interface between epithelium and cortical fibres and changes in adhesion to the capsule, respectively. High levels of betacatenin mRNA and protein were present in all lens types. However, with increasing complexity of alphaSMA organisation, betacatenin staining disappeared from the cell margin and basal infoldings and was shifted towards the cytoplasm and nucleus. The presence of alphaSMA in DON, the absence of any correlation between mRNA level and age, and the modest increase in complexity of alphaSMA-containing structures in CAT argue against an inevitable link between alphaSMA expression and the development of age-related cataract. Low levels of alphaSMA expression may reflect repair of normal wear and tear. In pathologic situations such as PSO and ASF, persisting stimulation and additional incentives may induce increased alphaSMA expression and more elaborate patterning, eventually leading to completion of EMT.

African swine fever (ASF) virus production was inhibited more than 100 fold by 5 mM glucosamine, 2 mM 2-deoxyglucose and 3 microM tunicamycin. ASF virus induced in Vero cells the synthesis of 19 glycosylated components of molecular weights ranging from 9K to 220K, the major ones being those of 9K, 13K, 14K, 74K and 220K. At least five of the induced glycosylated components, of molecular weights 13K, 33K, 34K, 38K and 220K, were probably virus-coded glycoproteins, as suggested by a comparative analysis of the time course of synthesis and the antigenicity of these components in extracts from [35S]methionine or [14C]glucosamine-labeled infected cells. The non-protein glycosylated components present in extracellular ASF virus particles had a cellular origin.

Many of the human myometrial proteins associated with uterine quiescence and the switch to coordinated contractions at the onset of labor exist as alternatively spliced isoforms. There is now extensive evidence to indicate that the nuclear concentrations of the trans-acting splicing regulators SF2/ASF and hnRNP A1/A1B are fundamental in regulating the expression of specific protein isoforms derived from alternative splicing of single precursor messenger ribonucleic acid transcripts. The question thus arose as to whether these factors were also involved in regulating the expression of specific myometrial protein species within different uterine regions during human gestation and parturition. SF2/ASF and hnRNP A1/A1B expression was therefore determined in paired upper (corpus) and lower segment myometrial samples taken from individual women at term/during spontaneous labor and compared with nonpregnant control samples using specific monoclonal antibodies. We report that SF2/ASF levels were substantially increased in the lower uterine region, and this was associated with a parallel decrease in levels of hnRNP A1/A1B during gestation. Conversely, the opposite pattern was observed within the upper uterine region during pregnancy, where hnRNP A1/A1B was significantly up-regulated and SF2/ASF levels were much less than those found in the lower uterine segment. The differential expression of hnRNP A1/A1B and SF2/ASF in the upper and lower uterine segments may have a primary role in defining the formation of specific myometrial protein species associated with the known contractile and relaxatory properties of these regions before and during parturition.

Previous studies have shown that protein-protein interactions among splicing factors may play an important role in pre-mRNA splicing. We report here identification and functional characterization of a new splicing factor, Sip1 (SC35-interacting protein 1). Sip1 was initially identified by virtue of its interaction with SC35, a splicing factor of the SR family. Sip1 interacts with not only several SR proteins but also with U1-70K and U2AF65, proteins associated with 5' and 3' splice sites, respectively. The predicted Sip1 sequence contains an arginine-serine-rich (RS) domain but does not have any known RNA-binding motifs, indicating that it is not a member of the SR family. Sip1 also contains a region with weak sequence similarity to the Drosophila splicing regulator suppressor of white apricot (SWAP). An essential role for Sip1 in pre-mRNA splicing was suggested by the observation that anti-Sip1 antibodies depleted splicing activity from HeLa nuclear extract. Purified recombinant Sip1 protein, but not other RS domain-containing proteins such as SC35, ASF/SF2, and U2AF65, restored the splicing activity of the Sip1-immunodepleted extract. Addition of U2AF65 protein further enhanced the splicing reconstitution by the Sip1 protein. Deficiency in the formation of both A and B splicing complexes in the Sip1-depleted nuclear extract indicates an important role of Sip1 in spliceosome assembly. Together, these results demonstrate that Sip1 is a novel RS domain-containing protein required for pre-mRNA splicing and that the functional role of Sip1 in splicing is distinct from those of known RS domain-containing splicing factors.

BACKGROUND

The side effects of chlorhexidine (CHX) have stimulated the search for alternative antiplaque agents such as amine fluoride/stannous fluoride (ASF) and essential oils (EO). The aim of the study was to investigate the plaque-inhibiting effects of two commercially available mouthrinses containing ASF and EO, respectively.

METHODS

The study was an observer-masked, randomized, 5 x 5 Latin square cross-over design, balanced for carryover effects, involving 15 volunteers in a 4-day plaque regrowth model. A 0.12% CHX rinse and a saline solution served as positive and negative controls, respectively. On day 1, subjects received professional prophylaxis, suspended oral hygiene measures, and commenced rinsing with their allocated rinses. On day 5, subjects were scored for disclosed plaque. The ASF rinse was tested at two dosages: 10 and 20 ml (ASF-10 and ASF-20, respectively).

RESULTS

The ASF and EO rinses showed a significant inhibition of plaque regrowth compared to saline (P <0.0001), but the lowest plaque indices were obtained with the CHX product (P <0.01). There were no significant differences among products containing ASF-10, ASF-20, and EO (P >0.05). There was no correlation between the occurrence of side effects and the use of a particular rinse product (P >0.2).

CONCLUSIONS

ASF and EO mouthrinses exerted effective and similar plaque inhibition. The two dosages tested for ASF did not differ in plaque reduction. These findings, together with those from long-term trials, suggest that ASF and EO rinses may represent effective alternatives to CHX rinse as adjuncts to oral hygiene.

Human DNA topoisomerase I not only has DNA relaxing activity, but also splicing factors phosphorylating activity. Topo I shows strong preference for ATP as the phosphate donor. We used photoaffinity labeling with the ATP analogue [alpha-32P] 8-azidoadenosine-5'-triphosphate combined with limited proteolysis to characterize Topo I domains involved in ATP binding. The majority of incorporated analogue was associated with two fragments derived from N-terminal and C-terminal regions of Topo I, respectively. However, mutational analysis showed that deletion of the first 138 N-terminal residues, known to be dispensable for topoisomerase activity, did not change the binding of ATP or the kinase activity. In contrast, deletion of 162 residues from the C-terminal domain was deleterious for ATP binding, kinase and topoisomerase activities. Furthermore, a C-terminal tyrosine 723 mutant lacking topoisomerase activity is still able to bind ATP and to phosphorylate SF2/ASF, suggesting that the two functions of Topo I can be separated. These findings argue in favor of the fact that Topo I is a complex enzyme with a number of potential intra-cellular functions.

METHODS

Retrospective analysis of prospectively collected data.

OBJECTIVE

To compare the relative rates of pulmonary recovery and maximal pulmonary function with surgical approach.

BACKGROUND

Anterior versus posterior spinal fusion (ASF, PSF) for the treatment of adolescent idiopathic scoliosis (AIS) has been debated. Although procedures that violate the chest wall may compromise pulmonary function, lung function continues to improve after surgery at variable rates depending upon surgical approach.

METHODS

We reviewed the medical records from one hundred fifty nine AIS patients (age 15.6±2.2; 113 women; 46 men) treated with spinal fusion from 2003 to 2007 by a single surgeon. Forced expiratory volume in one second (FEV1), forced vital capacity (FVC), and radiographic measurements were evaluated before surgery and at 1, 3, 6, 12, and 24-months follow-up on average. Four surgical groups were compared: PSF, ASF (open thoracoabdominal approach for thoracolumbar curvature), video-assisted thoracoscopic surgical release with instrumentation (VATS-I), and VATS with PSF. FEV1 and FVC were fitted to model to evaluate the immediate postoperative pulmonary function (Yo), maximal recovery (Plateau), and rate (K) of pulmonary improvement.

RESULTS

Patients in each surgical subgroup were as follows: PSF (Lenke 1: n=50, Lenke 2,3: n=20), ASF (Lenke 5, n=35), VATS-I (Lenke 1=31, Lenke 3=1), and VATS+PSF (Lenke1: n=9, Lenke 2-6: n=13). Early postoperative pulmonary function was higher with ASF and PSF as compared to both VATS groups (P<0.05). Comparing all curve types, VATS-I showed a small decline of absolute FEV1 compared to PSF at 2-years follow-up. Comparing thoracic curves, however, no differences in FEV1 or FVC were noted at 6 to 12 months until 2-years follow-up. The rate of recovery (K) was equivalent for all surgical approaches and curve types.

CONCLUSIONS

Compared to ASF or PSF, VATS procedures showed an initial decline in pulmonary function, which resolved fully by 6- to 12-months follow-up. Modest declines in maximal pulmonary function with VATS-I were seen when comparing all curve types together but not when comparing Lenke 1 curves alone. VATS procedures for thoracic scoliosis and open approaches for thoracolumbar curve types were associated with minimal to no permanent deficits.

The present study examines the effect of animal-source-food (ASF) intake on arm muscle area growth as part of a larger study examining causal links between ASF intake, growth rate, physical activity, cognitive function and micronutrient status in Kenyan schoolchildren. This randomised, controlled feeding intervention study was designed with three isoenergetic feeding interventions of meat, milk, and plain traditional vegetable stew (githeri), and a control group receiving no snack. A total of twelve elementary schools were randomly assigned to interventions, with three schools per group, and two cohorts of 518 and 392 schoolchildren were enrolled 1 year apart. Children in each cohort were given feedings at school and studied for three school terms per year over 2 years, a total of 9 months per year: cohort I from 1998 to 2000 and cohort II from 1999 to 2001. Food intake was assessed by 24 h recall every 1-2 months and biochemical analysis for micronutrient status conducted annually (in cohort I only). Anthropometric measurements included height, weight, triceps skinfold (TSF) and mid-upper-arm circumference (MUAC). Mid-upper-arm muscle area (MAMA) and mid-upper-arm fat area (MAFA) were calculated. The two cohorts were combined for analyses. The meat group showed the steepest rates of gain in MUAC and MAMA over time, and the milk group showed the next largest significant MUAC and MAMA gain compared with the plain githeri and control groups (P< 0.05). The meat group showed the least increase in TSF and MAFA of all groups. These findings have implications for increasing micronutrient intake and lean body mass in primary schoolchildren consuming vegetarian diets.

Two methods were evaluated for the inactivation of African swine fever (ASV) and swine vesicular disease (SVD) viruses in pig slurry: chemical treatment and heat treatment. The addition of NaOH or Ca(OH)2 at different concentration/time combinations at 4 degrees C and 22 degrees C was examined, as was virus stability at different temperature/time combinations. ASF virus (ASFV) was less resistant to both methods than SVD virus (SVDV). In slurry from one source, ASFV was inactivated at 65 degrees C within 1 min, whereas SVDV required at least 2 min at 65 degrees C. However, it was found that thermal inactivation depended on the characteristics of the slurry used. Addition of 1% (w/v) of NaOH or Ca(OH)2 caused the inactivation of ASFV within 150 s at 4 degrees C; 0.5% (w/v) NaOH or Ca(OH)2 required 30 min for inactivation. NaOH or Ca(OH)2 (1% (w/v)) was not effective against SVDV at 22 degrees C after 30 min, and 1.5% (w/v) NaOH or Ca(OH)2 caused inactivation of SVDV at both 4 degrees C and 22 degrees C. At higher chemical concentrations or temperatures, ASFV and SVDV inactivation was faster in slurry than in buffered medium.

African swine fever (ASF) in wild boar emerged in Estonia for the first time in September 2014. The first affected region was located in the South of Estonia close to the border with Latvia. It was considered to be epidemiologically connected to the outbreaks in the North of Latvia. About two weeks later, cases were detected in the North of Estonia, close to the Russian border. In the present study, we aimed to investigate the epidemiological courses of the disease in the South and in the North of Estonia. Potential associations between risk factors and the laboratory test results for ASF were examined. A hierarchical Bayesian space-time model was used to analyze the temporal trend of the ASF seroprevalence in the two areas. Young wild boar were statistically significant more likely to be ASF-positive by both, serology and virus detection, than older animals. A statistically significant difference between the two areas in the temporal course of the seroprevalence was found. While the seroprevalence clearly increased in the South, it remained relatively constant in the North. These findings led to the hypothesis that ASF might have been introduced earlier into the North of Estonia then into the South of the country.

BACKGROUND

Variations and defects in alternative splicing are well known to be associated with a variety of human diseases and the stress response. We previously reported a decrease in glucocorticoid receptor (GR) alpha, but not GRbeta in mood disorder patients, suggesting an aberrant alternative splicing mechanism. To examine whether altered RNA splicing may underlie the pathophysiology of mood disorder, we evaluated the expression of a variety of SR protein splicing factors, a family of proteins indispensable for proper alternative splicing, in mood disorder patients.

METHODS

We used quantitative real-time PCR to measure expressions of SRp20, SRp30c, SC35, SRp40, SRp46, SRp54, SRp55, SRp75, ASF/SF2, and 9G8 mRNA in peripheral white blood cells of 33 mood disorder patients during a depressive episode. In addition, the expressions of SRp20 and SC35 mRNA were quantified for 78 mood disorder patients in a remissive state, and 32 the first-degree relatives of these mood disorder patients.

RESULTS

A significant correlation was observed between SRp30c and the GRbeta/GRalpha ratio in control subjects, but not in mood disorder patients. Increased expression of SRp20 but not SRp30c mRNA was observed in bipolar disorder patients in both the depressive and remissive states. Major depressive disorder patients did not show any significant change in mRNA levels of SR proteins.

CONCLUSIONS

Subjects were Japanese adults. Patient treatment was not standardized.

CONCLUSIONS

These results suggest that aberrant alternative splicing machinery caused by increased SRp20 mRNA expression would be associated with the pathophysiology of bipolar disorder.

The human papillomavirus (HPV) life cycle is tightly linked to differentiation of the infected epithelial cell, suggesting a sophisticated interplay between host cell metabolism and virus replication. Previously, we demonstrated in differentiated keratinocytes in vitro and in vivo that HPV type 16 (HPV16) infection caused increased levels of the cellular SR splicing factors (SRSFs) SRSF1 (ASF/SF2), SRSF2 (SC35), and SRSF3 (SRp20). Moreover, the viral E2 transcription and replication factor that is expressed at high levels in differentiating keratinocytes could bind and control activity of the SRSF1 gene promoter. Here, we show that the E2 proteins of HPV16 and HPV31 control the expression of SRSFs 1, 2, and 3 in a differentiation-dependent manner. E2 has the greatest transactivation effect on expression of SRSF3. Small interfering RNA depletion experiments in two different models of the HPV16 life cycle (W12E and NIKS16) and one model of the HPV31 life cycle (CIN612-9E) revealed that only SRSF3 contributed significantly to regulation of late events in the virus life cycle. Increased levels of SRSF3 are required for L1 mRNA and capsid protein expression. Capsid protein expression was regulated specifically by SRSF3 and appeared independent of other SRSFs. Taken together, these data suggest a significant role of the HPV E2 protein in regulating late events in the HPV life cycle through transcriptional regulation of SRSF3 expression.

Human papillomavirus replication is accomplished in concert with differentiation of the infected epithelium. Virus capsid protein expression is confined to the upper epithelial layers so as to avoid immune detection. In this study, we demonstrate that the viral E2 transcription factor activates the promoter of the cellular SRSF3 RNA processing factor. SRSF3 is required for expression of the E4(^)L1 mRNA and so controls expression of the HPV L1 capsid protein. Thus, we reveal a new dimension of virus-host interaction crucial for production of infectious virus. SRSF proteins are known drug targets. Therefore, this study provides an excellent basis for developing strategies to regulate capsid protein production in the infected epithelium and the production of new virions.

Animal source foods (ASF) provide critical micronutrients in highly bioavailable forms, with the potential to efficiently address undernutrition among young children living in developing countries. There is limited evidence for how livestock ownership might increase ASF intake in poor households either through own-consumption or income generation. Along with lack of nutrition knowledge, gender dimensions may affect the pathways leading from livestock ownership to child ASF intake and ultimately to young child growth. Using data from a large-scale impact evaluation conducted in Kenya, this study tested the hypothesis that co-owned/female-owned livestock would be associated with improved child growth, mediated by increases in ASF consumption. Data were collected from September 2010 to January 2011 from households in six provinces in Kenya on a broad range of agricultural, economic, social, health and nutrition factors. Children ages 6-60 months were included in this analysis (n = 183). In this sample, co-owned/female-owned livestock was valued at 18,861 Kenyan shillings in contrast with male-owned livestock valued at 66,343 Kenyan shillings. Multivariate linear regression models showed a positive association between co-owned/female-owned livestock with child weight-for-age z score (WAZ) after adjusting for caregiver education level, income, child age, and child sex. A mediating effect by child ASF intake was evident, explaining 25% of the relationship of livestock ownership with child WAZ, by Sobel-Goodman test (p < .05). A trend towards significance was demonstrated for co-owned/female-owned livestock and height-for-age z score (HAZ), and no effect was apparent for weight-for-height z score (WHZ). The partial mediating effect may be indicative of other factors inherent in co-owned/female-owned livestock such as higher status of females in these households with greater influence over other child care practices promoting growth. Nonetheless, our study suggests targeting females in livestock production programming may better ensure improvements in child nutrition.

The expanding distribution of African swine fever (ASF) is threatening the pig industry worldwide. Most outbreaks occur in backyard and small-scale herds, where poor farmers often attempt to limit the disease's economic consequences by the emergency sale of their pigs. The risk of African swine fever virus (ASFV) release via this emergency sale was investigated. Simulation modeling was used to study ASFV transmission in backyard and small-scale farms as well as the emergency sale of pigs, and the potential impact of improving farmers and traders' clinical diagnosis ability-its timeliness and/or accuracy-was assessed. The risk of ASFV release was shown to be high, and improving farmers' clinical diagnosis ability does not appear sufficient to effectively reduce this risk. Estimates obtained also showed that the distribution of herd size within the backyard and small-scale sectors influences the relative contribution of these farms to the risk of release of infected pigs. These findings can inform surveillance and control programs.

Human interferon alpha (IFN-alpha) and interferon gamma (IFN-gamma) inhibit African swine fever (ASF) virus replication in Vero cells. IFN-alpha and IFN-gamma exert a synergistic inhibition. Human tumor necrosis factor (TNF) does not inhibit ASF virus replication in this cell line, but in combination with IFNs it has antiviral enhancing activity. Analysis of the mechanism of inhibition suggests that the action of these cytokines blocks a step that comes prior to DNA replication. The 2'-5' A synthetase activity is induced in Vero cells by treatment with these cytokines and is activated after ASF virus infection. More interesting is the finding that continuous treatment with IFN-alpha cures Vero cells from lytic and persistent infections with ASF virus. A potential application of IFN for the treatment of animals carrying the virus is suggested.

African swine fever (ASF) is caused by African swine fever virus (ASFV), a tick-borne DNA virus. Soft ticks of the genus Ornithodoros are the only biological vectors of ASFV recognized so far. Although other hard ticks have been tested for vector competence, two commonly found tick species in Europe, Ixodes ricinus and Dermacentor reticulatus, have not been assessed for their vector competence for ASFV. In this study, we aimed to determine whether virus replication can occur in any of these two hard tick species (I. ricinus and/or D. reticulatus), in comparison with O. moubata (the confirmed vector), after feeding them blood containing different ASFV isolates using an improved in vitro system. DNA quantities of ASFV in these infected hard ticks were measured systematically, for 6 weeks in I. ricinus, and up to 8 weeks in D. reticulatus, and the results were compared to those obtained from O. moubata. There was evidence of virus replication in the O. moubata ticks. However, there was no evidence of virus replication in I. ricinus or D. reticulatus, even though viral DNA could be detected for up to 8 weeks after feeding in some cases. This study presents the first results on the possible vector competence of European hard (ixodid) ticks for ASFV, in a validated in vitro feeding setup. In conclusion, given the lack of evidence for virus replication under in vitro conditions, D. reticulatus and I. ricinus are unlikely to be relevant biological vectors of ASFV.

The isolation of 98/ASF/NG, a strain of African Swine Fever Virus (ASFV) associated with a 1998 epizootic in Nigeria, is reported. This first isolate of the virus from West Africa was identified through a successful polymerase chain reaction (PCR) amplification and sequencing of a 280 base pair (bp) fragment of the Major Capsid Protein (VP72) gene. Further amplification and sequence analysis of a 1.9 kilobase pair (kbp) fragment encompassing the complete VP72 gene showed that the isolate has a 92.2%, 92.4%, and 97.2% homology with previously sequenced Ugandan, Dominican Republican and Spanish isolates respectively. Of the 50 nucleotide changes observed in this highly conserved gene, 45 were found to result in 40 amino acid changes clustered around the central region (position 426 to 516) of the VP 72 protein while changes at the remaining 5 positions were silent. These changes also led to the loss of two out of the seven potential N-glycosylation sites which are in this gene conserved among all isolates. The possible epizootiological implications of such mutations in a highly conserved gene of a DNA virus is discussed in relation to this outbreak.

In vivo effective relaxation rates in normal rat liver were evaluated for four dextran coated iron oxide agents: monocrystalline iron oxide nanocolloid (MION) with a mean particle diameter of 3.9 nm, a polycrystalline agent (PION) with a larger mean diameter of 12 nm, and these two agents labeled with the asialofetuin (ASF) protein for high hepatocytic receptor binding affinity (MION-ASF and PION-ASF). Using echo planar imaging at 2 Tesla, dose response was measured with measured with high temporal resolution for 3 h after injection of agent, and by comparing with relaxivities in vitro and in brain, dominant in vivo contrast phenomena were elucidated. While transverse relaxivity for PION-ASF exceeded that for MION-ASF by almost a factor of 2 in solution, relaxation rates in vivo became equivalent. Liver relaxation using non-ASF agents was consistent with rapid water exchange between vascular and extravascular compartments, which dominated relaxation as a result of agent accumulation in Kupffer cells.

African swine fever virus (ASFV) causes high morbidity and mortality in swine (Sus scrofa), for which there is no commercially available vaccine. Recent outbreaks of the virus in Trans-Caucasus countries, Eastern Europe, Belgium and China highlight the urgent need to develop effective vaccines against ASFV. Previously, we evaluated the immunogenicity of a vaccination strategy designed to test various combinations of ASFV antigens encoded by DNA plasmids and recombinant proteins with the aim to activate both humoral and cellular immunity. Based on our previous results, the objective of this study was to test the combined DNA-protein vaccine strategy using a cocktail of the most immunogenic antigens against virulent ASFV challenge. Pigs were vaccinated three times with a cocktail that included ASFV plasmid DNA (CD2v, p72, p32, +/-p17) and recombinant proteins (p15, p35, p54, +/-p17). Three weeks after the third immunization, all pigs were challenged with the virulent ASFV Armenia 2007 strain. The results showed that vaccinated pigs were not protected from ASFV infection or disease. Compared to the non-vaccinated controls, earlier onset of clinical signs, viremia, and death were observed for the vaccinated animals following virulent ASFV challenge. ASFV induced pathology was also enhanced in the vaccinated pigs. Furthermore, while the vaccinated pigs developed antigen-specific antibodies, immunized pig sera at the time of challenge lacked the capacity to neutralize virus, and instead was observed to enhance ASFV infection in vitro. The results of this work points to a putative immune enhancement mechanism involved in ASFV pathogenesis that warrants further investigation. This pilot study provides insight for the selection of appropriate combinations of ASFV antigens for the development of a rationally-designed, safe, and efficacious vaccine for ASF.

African swine fever (ASF) is currently spreading westwards throughout Europe and eastwards into China, with cases occurring in both wild boar and domestic pigs. A generic risk assessment framework is used to determine the probability of first infection with ASF virus (ASFV) at a fine spatial scale across European Union Member States. The framework aims to assist risk managers across Europe with their ASF surveillance and intervention activities. Performing the risk assessment at a fine spatial scale allows for hot-spot surveillance, which can aid risk managers by directing surveillance or intervention resources at those areas or pathways deemed most at risk, and hence enables prioritization of limited resources. We use 2018 cases of ASF to estimate prevalence of the disease in both wild boar and pig populations and compute the risk of initial infection for 2019 at a 100 km² cell resolution via three potential pathways: legal trade in live pigs, natural movement of wild boar, and legal trade in pig meat products. We consider the number of pigs, boar and amount of pig meat entering our area of interest, the prevalence of the disease in the origin country, the probability of exposure of susceptible pigs or boar in the area of interest to introduced infected pigs, boar, or meat from an infected pig, and the probability of transmission to susceptible animals. We provide maps across Europe indicating regions at highest risk of initial infection. Results indicate that the risk of ASF in 2019 was predominantly focused on those regions which already had numerous cases in 2018 (Poland, Lithuania, Hungary, Romania, and Latvia). The riskiest pathway for ASFV transmission to pigs was the movement of wild boar for Eastern European countries and legal trade of pigs for Western European countries. New infections are more likely to occur in wild boar rather than pigs, for both the pig meat and wild boar movement pathways. Our results provide an opportunity to focus surveillance activities and thus increase our ability to detect ASF introductions earlier, a necessary requirement if we are to successfully control the spread of this devastating disease for the pig industry.

Plant and animal lectins bind and cross-link certain multiantennary oligosaccharides, glycopeptides, and glycoproteins. This can lead to the formation of homogeneous cross-linked complexes, which may differ in their stoichiometry depending on the nature of the sugar receptor involved. As a precisely defined ligand, we have employed bovine asialofetuin (ASF), a glycoprotein that possesses three asparagine-linked triantennary complex carbohydrate chains with terminal LacNAc residues. In the present study, we have compared the carbohydrate cross-linking properties of two Lac-specific plant lectins, an animal lectin and a naturally occurring Lac-binding polyclonal immunoglobulin G subfraction from human serum with the ligand. Quantitative precipitation studies of the Lac-specific plant lectins, Viscum album agglutinin and Ricinus communis agglutinin, and the Lac-specific 16 kDa dimeric galectin from chicken liver demonstrate that these lectins form specific, stoichiometric cross-linked complexes with ASF. At low concentrations of ASF, 1:9 ASF/lectin (monomer) complexes formed with both plant lectins and the chicken lectin. With increasing concentrations of ASF, 1:3 ASF/lectin (monomer) complexes formed with the lectins irrespective of their source or size. The naturally occurring polyclonal antibodies, however, revealed a different cross-linking behavior. They show the formation of 1:3 ASF/antibody (per Fab moiety) cross-linked complexes at all concentrations of ASF. These studies demonstrate that Lac-specific plant and animal lectins as well as the Lac-binding immunoglobulin subfraction from specific stoichiometric cross-linked complexes with ASF. These results are discussed in terms of the structure-function properties of multivalent lectins and antibodies.

BACKGROUND

The aim of this study was to evaluate the clinical and antibacterial properties of alcohol-free mouthrinses, an amine fluoride/stannous fluoride mouthrinse (ASF) and two triclosan solutions in comparison with a chlorhexidine and a placebo rinse.

METHODS

In a double-blind, randomised 5-cell cross-over 4-day plaque regrowth study, 19 volunteers rinsed 2 x a day with 15 ml of each of the distributed solutions. Each test cycle was followed by a 10 days wash-out period. On day 0 of each test week, volunteers received a dental prophylaxis. Thereafter they refrained from all mechanical oral hygiene procedures for the next four days. Plaque regrowth was assessed daily by the plaque index and on day 4 by calculating the plaque area with a computer program after disclosure and photography of the front teeth. The vitality of the plaque was examined on days 1 to 4 by the vital fluorescence technique.

RESULTS

19 participants completed the study. Compared to the placebo the ASF solution showed 15.7% (p>0.5), 30.6% (p<0.001), 40.5% (p<0.001) and 44.7% (p<0.001) reductions on the consecutive days 14 in plaque index and a reduction of 61.9% in plaque area. The decrease of vitality of supragingival plaque was highly significant compared to placebo on every test day ranging between 30.9% and 36.6%. A reduction in plaque index from 19.4% (p<0.01) on day 2, 34.9% (p<0.001) on day 3 and 40.4% (p<0.001) on day 4 concommitant with a reduction in plaque area of 48.9% (p<0.001) was noted for alcohol-free chlorhexidine. Concerning the vitality chlorhexidine reduced the percentage of vital bacteria significantly on every day (16.0% to 24.9%). The reductions in mean plaque index for the 0.15% triclosan solution during the test period were 6.3%, 22.4%, 24.6% and 36% and in plaque area 41.8%. Vitality was significantly reduced at every day compared to placebo. Plaque Index reduction with the 0.02% triclosan increased from 15.3% (day 2) to 31.0% (day 4) and a reduction of 23.6% was seen in plaque area. A significant effect concerning the vitality of the plaque was only found at the first and the last day of the test cycle.

CONCLUSIONS

Alcohol-free mouthrinse solutions were shown to be effective in reducing both plaque accumulation and plaque biofilm vitality compared to a placebo solution.