Acrometastasis in Breast Carcinoma.

Background: Metastasis to the bone in breast cancer patients is common, but metastasis specifically to the appendicular skeleton is rare. A limited number of cases in the literature describe metastatic breast cancer to the distal limbs, also known as acrometastasis. Acrometastasis in a patient with breast cancer should prompt evaluation for diffuse metastatic disease. Case Report: We describe the case of a patient with recurrent triple-negative metastatic breast cancer who presented with thumb pain and swelling. Radiograph of the hand demonstrated focal soft tissue swelling over the first distal phalanx with erosive changes to the bone. Palliative radiation to the thumb resulted in symptom improvement. However, the patient succumbed to widespread metastatic disease. At autopsy, the thumb lesion was confirmed as metastatic breast adenocarcinoma. Conclusion: Metastatic breast carcinoma to the distal appendicular skeleton, specifically to the first digit, is a rare presentation of bony metastasis and can be an indication of late, widespread disease.

The Use of Phage Display and Yeast Based Expression System for the Development of a Von Willebrand Factor Propeptide Assay: Development of a Von Willebrand Factor Propeptide Assay.

BACKGROUND: The diagnosis of von Willebrand disease is complex due to the heterogeneity of the disease. About eighty percent of von Willebrand disease patients are diagnosed with a quantitative defect of von Willebrand factor (VWF) where fifty percent is due to an increased clearance of von Willebrand factor. These patients do not respond well to the treatment of choice, Desmopressin (DDAVP) due to decreased efficacy. The ratio between the VWF propeptide and the mature VWF antigen is used to diagnose these patients. Commercial VWF propeptide assays are too expensive for use in developing countries. In this study, we developed a cost-effective ELISA assay. METHODS: We first displayed VWF propeptide on yeast. Antibody fragments were selected against the displayed VWF propeptide by using phage display technology. The antibodies were used to develop a cost-effective VWF propeptide assay and compared to a commercial VWF propeptide assay. RESULTS: Two of these antibody fragments bound specific to the VWF propeptide and not to the yeast used for the expression of the propeptides. These purified antibody fragments were able to detect VWF propeptide in normal plasma. CONCLUSION: Our assay performed well when compared to a commercial kit. It also showed a higher binding affinity for VWF propeptide in plasma at especially lower plasma concentrations.

Potential Treatment Options in a Post-antibiotic Era.

Following the Golden Age of antibiotic discovery in the previous century, the rate of antibiotic discovery has plummeted during the past 50 years while the incidence of antimicrobial resistance is ever-increasing. Presently, humankind is forced to address a major public health threat in the form of multiple drug resistance and urgent action is required to halt the advent of a post-antibiotic era. This chapter aims to draw the attention to the escalating global crisis of antimicrobial resistance fueled by the irresponsible use of antibiotics in healthcare and animal production sectors. The merits of alternative prevention and treatment options, including vaccines, herbal products, bacteriophages, and improved biosecurity measures are also discussed.

Lactococcus garvieae: an emerging bacterial pathogen of fish.

Lactococcus garvieae is the causative agent of lactococcosis, a hyperacute, haemorrhagic septicaemia of fish. This bacterium is also considered an emerging zoonotic pathogen, as reports of human infection are increasing. Significant economic loss in aquaculture is suffered as a result of lactococcosis, as numerous freshwater and marine species of commercial interest are affected. Development of antibiotic resistance in L. garvieae to several chemotherapeutic agents complicates and restricts treatment options. Effective, sustainable treatment and prevention options are thus needed, but progress is impeded by the lack of knowledge concerning several aspects of the disease and the pathogen. This review aims to present the latest research on L. garvieae, with specific focus on pathogenesis, virulence factors, risks associated with chemotherapeutic administration and possible control options.

Abstracts of the 24th international isotope society (UK group) symposium: synthesis and applications of labelled compounds 2015.

The 24th annual symposium of the International Isotope Society's United Kingdom Group took place at the Møller Centre, Churchill College, Cambridge, UK on Friday 6th November 2015. The meeting was attended by 77 delegates from academia and industry, the life sciences, chemical, radiochemical and scientific instrument suppliers. Delegates were welcomed by Dr Ken Lawrie (GlaxoSmithKline, UK, chair of the IIS UK group). The subsequent scientific programme consisted of oral presentations, short 'flash' presentations in association with particular posters and poster presentations. The scientific areas covered included isotopic synthesis, regulatory issues, applications of labelled compounds in imaging, isotopic separation and novel chemistry with potential implications for isotopic synthesis. Both short-lived and long-lived isotopes were represented, as were stable isotopes. The symposium was divided into a morning session chaired by Dr Rebekka Hueting (University of Oxford, UK) and afternoon sessions chaired by Dr Sofia Pascu (University of Bath, UK) and by Dr Alan Dowling (Syngenta, UK). The UK meeting concluded with remarks from Dr Ken Lawrie (GlaxoSmithKline, UK).

Control of eight predominant Eimeria spp. involved in economic coccidiosis of broiler chicken by a chemically characterized essential oil.

AIM: To control eight most predominant Eimeria spp. involved in the economic disease of coccidiosis in broiler chicken, by a chemically characterized essential oil of eucalyptus and peppermint. METHODS AND RESULTS: The experimental design consisted of 160 day-old-broiler chicks, divided into four equal groups (G1 , G2 , G3 and G4 ), with 40 birds per group. Each group was divided into four equal subgroups. Birds in G1 were deprived of essential oil treatment and of Eimeria challenge. Birds in G2 were unchallenged, and administered the essential oil in drinking water at 0.69 ml kg(-1) body weight. Birds in G3 were untreated with essential oil, and each of its four subgroups was challenged at a different age (14, 21, 28 and 35 days). Birds in G4 were treated with essential oil, and challenged in the same manner as for G3 . Equal number of birds from all subgroups (n = 10) were sacrificed at the sixth day after the time allocated for each challenge. The 6 day incubation period post challenge resulted in respective mean per cent weight increase in G2 and G1 birds equivalent to 57.8 and 53.1% (P < 0.05). In addition, the essential oil improved the per cent weight increase in challenged birds (54.6%) compared to the challenged-untreated birds (18.6%) (P < 0.05). The mean feed conversion, mortality, intestinal lesion scores and oocyst counts were significantly reduced in the challenged-treated birds compared to the challenged-untreated birds (P < 0.05). CONCLUSIONS: The results support the hypothesis of using the essential oils of eucalyptus and peppermint to control the most prevalent Eimeria spp. involved in coccidiosis of broiler chicken, helping in improvement of their production, alleviation of lesions and reduction in intestinal oocyst counts. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information about the possibility of using this blend of essential oil as a coccidiostat for the protection of broiler chickens against the prevalent eight Eimeria spp. of coccidiosis.

Understanding the impacts of care farms on health and well-being of disadvantaged populations: a protocol of the Evaluating Community Orders (ECO) pilot study.

INTRODUCTION: Care farms, where all or part of the farm is used for therapeutic purposes, show much potential for improving the health and well-being of a range of disadvantaged groups. Studies to date have been qualitative or observational, with limited empirical evidence of the effectiveness of care farms in improving health and well-being. Understanding the underlying mechanisms that lead to improvements for different disadvantaged groups is a further gap in the evidence. Participants in this study are offenders serving community orders. Their low socioeconomic status and poor health outcomes relative to the general population exemplifies disadvantage. METHODS AND ANALYSIS: This paper describes the protocol of a study to understand the impacts of care farms and to pilot the design and tools for a study to assess cost-effectiveness of care farms in improving the quality of life of offenders. As a pilot study, no power calculation has been conducted. However, 150 offenders serving community sentences on care farms and 150 on other probation locations (eg, litter picking, painting) will be recruited over a 1-year period. Changes in quality of life, measured by Clinical Outcome in Routine Evaluation-Outcome Measure, health and reconvictions of offenders at care farms compared to other probation locations will be analysed to inform the sample size calculation for the follow on study. The feasibility of recruitment, retention, collecting cost data and modelling cost-effectiveness will also be assessed. The study will use qualitative methods to explore the experiences of offenders attending care farms and perceptions of probation and care farm staff on the processes and impacts of the intervention. ETHICS AND DISSEMINATION: Findings will be published and inform development of a natural experiment and will be disseminated to probation services, care farms and academics. University of Leeds Ethical Review Board approved: SoMREC/13/014. National Offender Management Service (NOMS) approved: 2013-257.

Modulation by essential oil of vaccine response and production improvement in chicken challenged with velogenic Newcastle disease virus.

AIM: To evaluate the impact of Eucalyptus and peppermint essential oils on immune modulation and production of broiler chicken challenged with a molecularly characterized velogenic NewCastle disease virus (vNDV). METHODS AND RESULTS: The experimental design included five treatments with three replicate pens/treatment comprised of 12-day-old broilers chicks/replicate. The five treatments included a positive challenge control (non-NDV vaccinated/nonessential oil treated/challenged) (NNEOC), a negative challenge control (NDV vaccinated/essential oil treated/unchallenged) (VEOU), a non-NDV vaccinated/essential oil treated/challenged (NEOC), a NDV vaccinated/nonessential oil treated/challenged (VNEOC) and a NDV vaccinated/essential oil treated/challenged (VEOC). The lowest mean survival rate (0·0%) and lowest production performance were obtained by the positive challenge control, while the best mean survival (93·3%) and average body weight (2649 g) were obtained by the negative challenge controls (P < 0·05). Among the three others challenged treatments, the best mean survival (79·2%), highest mean body weight at 42 days of age (2445 g), the lowest feed conversion ratio (1·60) and the highest serum conversion immunopotentiation at 35 days of age determined by ELISA and hemagglutination titres were obtained by the VEOC birds compared with respective means obtained by birds of the NEOC and VNEOC treatments (P < 0·05). CONCLUSIONS: The results supported the possibility of using the essential oils of Eucalyptus and Peppermint in broilers to immunopotentiate the response to vaccination against velogenic NDV, helping in significant improvement of survival and production. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information about the potential use of essential oils of eucalyptus and peppermint that can be exploited as commercial immunopotentiators for the protection of NDV-vaccinated broiler chickens against economic velogenic NDV.

Multiplex polymerase chain reaction for screening avian pathogenic Escherichia coli for virulence genes.

Colibacillosis is a disease in poultry caused by avian pathogenic Escherichia coli (APEC) strains which leads to great economic losses in the poultry industry. These E. coli strains contain various virulence genes which grant the bacteria the ability to proliferate in the poultry host and cause disease. Many genes which can contribute to virulence have been identified and can be used to screen E. coli strains to infer pathogenicity and aid in the identification and classification of APEC. Multiplex polymerase chain reaction methods were designed and optimized to rapidly detect 18 different virulence genes in E. coli strains that were isolated in South Africa and Zimbabwe from various sources, including from chickens showing signs of colibacillosis. Virulence gene profiles were constructed for each E. coli isolate from the multiplex data for the comparison of the colibacillosis isolates with the other isolates. The South African E. coli isolated from chickens with signs of colibacillosis showed higher virulence gene prevalence in comparison with the Zimbabwean and other samples except those isolated from chicken faeces. The multiplex polymerase chain reaction designed in the present study successfully screened E. coli isolates for various APEC-related virulence genes, including genes recently described in the literature.

Study on the efficacy and safety of different antigens and oil formulations of infectious coryza vaccines containing an NAD-independent strain of Avibacterium paragallinarum.

The present study was designed to assess and compare three different formulations of the new Onderstepoort infectious coryza (IC) quadrivalent vaccine, which contain an NAD-independent strain of Avibacterium paragallinarum (previously known as Haemophilus paragallinarum), and a commercial IC vaccine, not containing an NAD-independent strain, for their safety and ability to protect chickens of varying ages against virulent challenges with four different serovars of A. paragallinarum, including the NAD-independent strain of the C-3 serovar. Four groups of 140 chickens each were vaccinated at the age of 17 weeks and revaccinated at the age of 19 weeks with each of the four vaccine formulations. A similar sized group of non-vaccinated chickens was used as control. Two rounds of challenge were conducted: a group of chicken in each vaccination group was challenged between 31 and 35 weeks of age, while another group was challenged between 51 and 55 weeks of age. The "in-contact" challenge model was used in this experiment. For each vaccination group, the four challenge strains representing four local serovars were used in each challenge round. The efficacy of the vaccines was compared based on overall protection levels obtained and the duration of protection. The safety of the different vaccines was determined by the severity of post-vaccination reactions. The need for the incorporation of the NAD-independent strain in the vaccine was evidenced by the low protection level against NAD-independent challenge recorded in the group of birds vaccinated with the commercial vaccine. The results obtained confirmed not only the variation in virulence of different South African serovars, with serovar C-3 being the most virulent and serovar B having almost no virulence but also the age related increase in susceptibility. The importance of a suitable formulation of the vaccine is discussed.

Haemophilus paragallinarum haemagglutinin: role in adhesion, serotyping and pathogenicity.

It is suggested that Haemophilus paragallinarum requires at least three haemagglutinins for adhesion during infection. This paper reports the partial purification and characterization of the HA-L haemagglutinin from H. paragallinarum strain 46-C3, a heat sensitive, trypsin sensitive haemagglutinin that has been shown to be the serovar specific haemagglutinin in this organism. Using the pl and molecular mass obtained, it was shown that this protein shares similarities with other types of adhesins found in Gram-negative bacteria. The haemagglutination assay conditions were optimized at pH 7.5 at 37 degrees C. It was also shown that activity is enhanced by the addition of Ca2+ and Mn2+ ions.

Genetic diversity of the Rep gene of beak and feather disease virus in South Africa.

A study on the genetic variation of Beak and feather disease virus (BFDV) isolates in South Africa was performed by amplifying and sequencing a region within the ORF1 of the genome. Six different BFDV isolates were found in 15 psittacine species from 6 regions within South Africa, representing three unique isolates and three isolates that clustered into a budgerigar lineage (BG) previously described.

Beak and feather disease virus haemagglutinating activity using erythrocytes from African Grey parrots and Brown-headed parrots.

Psittacine beak and feather disease (PBFD) is a common viral disease of wild and captive psittacine birds characterized by symmetric feather loss and beak deformities. The causative agent, beak and feather disease virus (BFDV), is a small, circular single-stranded DNA virus that belongs to the genus Circovirus. BFDV can be detected by PCR or the use of haemagglutination (HA) and haemagglutination inhibition (HI) assays that detect antigen and antibodies respectively. Erythrocytes from a limited number of psittacine species of Australian origin can be used in these tests. In South Africa, the high cost of these birds makes them difficult to obtain for experimental purposes. Investigation into the use of erythrocytes from African Grey parrots and Brown-headed parrots yielded positive results showing the haemagglutinating activity of their erythrocytes with purified BFDV obtained from confirmed clinical cases of the disease. The HA activity was further confirmed by the demonstration of HI using BFDV antiserum from three different African Grey parrots previously exposed to the virus and not showing clinical signs of the disease.

Effects of differences in virulence of different serovars of Haemophilus paragallinarum on perceived vaccine efficacy.

The virulence of four South African field isolates of NAD-dependent Haemophilus paragallinarum and two field isolates of NAD-independent H. paragallinarum has previously been tested in unvaccinated chickens. In this study, the disease profiles caused by the NAD-dependent isolates of H. paragallinarum in vaccinated chickens were studied. It was shown that the clinical signs induced in the vaccinated chickens were substantially less severe than were those in unvaccinated chickens, as was expected. However, due to the high virulence of the serovar C-3 isolates, clinical signs in the vaccinated chickens challenged with this isolate were still detected. These were as severe as those occurring in unvaccinated chickens challenged with serovar B-1 isolates. Although the clinical signs induced in unvaccinated birds challenged with serovar A-1 were more severe than those occurring when vaccinated birds were challenged with serovar C-3, the overall disease profiles were similar. Substantial clinical signs were recorded in vaccinated birds challenged with serovar C-3. This could be interpreted as vaccination failure if the disease profile obtained in unvaccinated birds is not considered. It was found that a high level of protection was provided by this vaccine against challenge by serovar C-3. The high virulence of this serovar resulted in the development of clinical signs in vaccinated birds. These findings could possibly explain the large number of so-called vaccination failures that are reported in South Africa.

The testing and modification of a commercially available transport medium for the transportation of pure cultures of Haemophilus paragallinarum for serotyping.

Haemophilus paragallinarum, the causative agent of infectious coryza in poultry, is an extremely fastidious organism requiring specific growth conditions for isolation. For complete control of the disease in regions where more that one of the serovars of the different serogroups occurs, it is essential that the bacterium causing the problem be isolated and serotyped. This work describes the modification and testing of transport media, which will ensure the survival of the causative agents in suspected infectious coryza cases for transport to a laboratory where the bacterium can be isolated and serotyped. The various transport media used are based on commercially available Amies Transport Medium supplemented with the different supplements used for the growth of H. paragallinarum. It was established that the bacterium remains viable for up to 18 days in Amies Transport Medium containing all the supplements when stored at 4 degrees C or 37 degrees C. At room temperature or 25 degrees C, there was no difference in the survival of H. paragallinarum in commercial Amies Transport Medium (without charcoal) and Amies Transport Medium with supplements.

Genetic organisation of the capsule transport gene region from Haemophilus paragallinarum.

The region involved in export of the capsule polysaccharides to the cell surface of Haemophilus paragallinarum was cloned and the genetic organisation determined. Degenerate primers designed from sequence alignment of the capsule transport genes of Haemophilus influenzae, Pasteurella multocida and Actinobacillus pleuropneumoniae were used to amplify a 2.6 kb fragment containing a segment of the H. paragallinarum capsule transport gene locus. This fragment was used as a digoxigenin labelled probe to isolate the complete H. paragallinarum capsule transport gene locus from genomic DNA. The sequence of the cloned DNA was determined and analysis revealed the presence of four genes, each showing high homology with known capsule transport genes. The four genes were designated hctA, B, C and D (for H. paragallinarum capsule transport genes) and the predicted products of these genes likely encode an ATP-dependent export system responsible for transport of the capsule polysaccharides to the cell surface, possibly a member of a super family designated ABC (ATP-binding cassette) transporters.

Limitation of the spread and impact of infectious coryza through the use of a continuous disinfection programme.

The effect of a continuous disinfection programme, using the non-toxic disinfectant Virukill, in layers, on the spread and impact of infectious coryza, caused by Haemophilus paragallinarum was evaluated. In this experiment, both unvaccinated layers and layers vaccinated against infectious coryza were used. Duplicate smaller groups of vaccinated and unvaccinated chickens were challenged with different serovars of both NAD-dependent as well as NAD-independent isolates of Haemophilus paragallinarum. One group of chickens challenged with each of the different becterial serovars was treated with the continuous disinfection programme, while the other group remained as the untreated controls. The clinical signs of infectious coryza were evaluated over a period of 20 days in each group. The egg production over this period was also evaluated. It was found in all experimental challenges, that the severity of the symptoms was reduced in the birds receiving the continuous disinfection programme. The drop in egg production was also found to be less severe in the treated groups when compared to the untreated control groups. The duration of infection was found to be either unchanged, or shorter in the birds treated with the continuous disinfection programme. In none of the experimental challenges was the duration or expression of clinical signs of IC increased due to the continuous disinfection programme.

Psittacine beak and feather disease virus in budgerigars and ring-neck parakeets in South Africa.

Psittacine beak and feather disease (PBFD) is a common disease of the psittacine species and is caused by the psittacine beak and feather disease virus (PBFDV). In this study the occurrence of the disease in ring-neck parakeets and budgerigars in South Africa suffering from feathering problems, using polymerase chain reaction as a diagnostic test was investigated. The genetic variation between viral isolates was also studied. Results indicate that PBFDV can be attributed to being the cause of feathering problems in some of the ring-neck parakeets and budgerigars in South Africa. Genetic variation of isolates occurs between species and individuals. A cheap and easy to use method of blood sample collection on filter paper for diagnostic purposes was also evaluated. It proved to be less stressful to the birds and did not inhibit further processes.