Serological and molecular survey of canine distemper virus in red foxes (Vulpes vulpes): Exploring cut-off values and the use of protein A in ELISA tests.

The wide distribution and ecological plasticity of the red fox (Vulpes vulpes) make it a potential reservoir for many infectious diseases shared with domestic and wild carnivores. One of such diseases is canine distemper, which is caused by an RNA virus and its main domestic reservoir is the dog. However, other carnivores can also participate in its maintenance, as shown by the recent upsurge of reported cases in wildlife in many parts of the world, and by the fact that red foxes may act as true reservoirs for canine distemper virus (CDV). The lack of validated serological tests for wildlife or other non-target species may be a handicap for monitoring this virus. In this study, serological assays were compared in 147 red fox sera using a commercial ELISA validated for its use in dogs and a non-specific modified ELISA with Protein A peroxidase conjugate to detect bound antibodies. In addition, the presence of CDV RNA in brain, spleen, lung, and liver samples from 144 foxes was investigated by a RT-qPCR. Through the comparison of the results of both ELISAs and the use of a finite mixture model of the optical density values obtained by both techniques, we adjusted the cut-off point of the commercial ELISA to obtain the seroprevalence in foxes. The overall seroprevalence detected was 53.7% (79/147) and 57.1% (84/147) by the commercial and modified ELISA, respectively, with a moderate agreement according to Cohen's Kappa statistic (κ = 0.491, z = 5.97, p < 0.0001). CDV RNA was detected in 30 out of 144 foxes, which resulted in 20.8% of CDV-infected foxes. At individual level, the results obtained by relating the serological status and the presence/absence of RNA in different organs were explained in terms of the pathogenesis of the infection. Our results highlight the convenience of adjusting the cut-off point when using an ELISA assay developed in domestic dogs for its use in foxes. Moreover, Protein A is confirmed to be a good alternative to be used in red foxes, presenting a good reactivity towards its IgG.

Disruption of the microbiota-gut-brain axis is a defining characteristic of the α-Gal A (-/0) mouse model of Fabry disease.

Fabry disease (FD) is an X-linked metabolic disease caused by a deficiency in α-galactosidase A (α-Gal A) activity. This causes accumulation of glycosphingolipids, especially globotriaosylceramide (Gb3), in different cells and organs. Neuropathic pain and gastrointestinal (GI) symptoms, such as abdominal pain, nausea, diarrhea, constipation, and early satiety, are the most frequent symptoms reported by FD patients and severely affect their quality of life. It is generally accepted that Gb3 and lyso-Gb3 are involved in the symptoms; nevertheless, the origin of these symptoms is complex and multifactorial, and the exact mechanisms of pathogenesis are still poorly understood. Here, we used a murine model of FD, the male α-Gal A (-/0) mouse, to characterize functionality, behavior, and microbiota in an attempt to elucidate the microbiota-gut-brain axis at three different ages. We provided evidence of a diarrhea-like phenotype and visceral hypersensitivity in our FD model together with reduced locomotor activity and anxiety-like behavior. We also showed for the first time that symptomology was associated with early compositional and functional dysbiosis of the gut microbiota, paralleled by alterations in fecal short-chain fatty acid levels, which partly persisted with advancing age. Interestingly, most of the dysbiotic features suggested a disruption of gut homeostasis, possibly contributing to accelerated intestinal transit, visceral hypersensitivity, and impaired communication along the gut-brain axis.

Influence of body lesion severity on oxidative status and gut microbiota of weaned pigs.

Body lesions in pigs are a common welfare concern, particularly during the weaning period. These lesions can lead to pain, infection, and impaired mobility, resulting in reduced growth performance and increased mortality. Moreover, weaning stress can affect gut microbiota, immune response and increase the oxidative stress of piglets during this transition period. It has been hypothesised that social stress and body lesions could contribute to affect the gut microbiota, physiological and immune response of piglets. The study aims to evaluate the impact of the body lesions due to social stress on microbial profile, immune response, and oxidative status of weaned piglets. Lesion score (LS) on skin, tail, ear, neck, middle trunk, and hind quarters was measured 1 week (28 days of age, T1) and 7 weeks postweaning (T2) on 45 tail-docked pigs according to the method suggested from the Walfer Quality® (2009) on a scale from 0 to 2. Based on the LS, at T1, piglets were classified as High LS (n = 16), when LS was >1 in at least two of the areas considered, or Low LS (n = 29). At T2, based on the same scoring system and to the LS observed at T1, piglets were divided into four groups: High to Low LS (H-L, n = 11), High to High LS (H-H, n = 5), Low to Low LS (L-L, n = 21) and Low to High LS (L-H, n = 8). Blood and faecal samples were collected at T1 and T2. At T1, pigs with a high LS had a lower biological antioxidant potential compared with the L group (P < 0.02). At T2, the L-H group had a lower Reactive Oxygen Metabolites concentration compared with the H-H group (P = 0.03) while the L-L group had a lower concentration of Immunoglobulin A compared with H-H and L-H groups (P = 0.02 and P = 0.04, respectively). At T1, piglets with high LS had a different microbiota compared to piglets with low LS (R2 = 0.04, P < 0.01). Low LS pigs were characterised by a higher abundance of Firmicutes, Blautia, Eubacterium coprostanoligenes, Faecalibacterium, Megasphaera, Subdoligranulum (P.adj < 0.05), while pigs with high LS were characterised by higher abundance of Bacteroidota, Rikenellaceae RC9, Prevotellaceae UCG-003, uncultured-Lachnospiraceae and uncultured-Oscillospiraceae (P.adj < 0.05). At T2, the H-H group were characterised by Oscillospirales-UCG-010, H-L by Agatobachter and L-L by Alloprevotella (P.adj < 0.05). Overall, this study provides valuable insights into the relationship between body lesions, oxidative stress, and gut microbiota in weaned pigs.

Gut microbiome structure and adrenocortical activity in dogs with aggressive and phobic behavioral disorders.

Accompanying human beings since the Paleolithic period, dogs has been recently regarded as a reliable model for the study of the gut microbiome connections with health and disease. In order to provide some glimpses on the connections between the gut microbiome layout and host behavior, we profiled the phylogenetic composition and structure of the canine gut microbiome of dogs with aggressive (n = 11), phobic (n = 13) and normal behavior (n = 18). Hormones' determination was made through Radio Immuno-Assay (RIA), and next generation sequencing of the V3-V4 gene region of the bacterial 16S rRNA was employed to determine gut microbiome composition. Our results did not evidence any significant differences of hormonal levels between the three groups. According to our findings, aggressive behavioral disorder was found to be characterized by a peculiar gut microbiome structure, with high biodiversity and enrichment in generally subdominant bacterial genera (i.e. Catenibacterium and Megamonas). On the other hand, phobic dogs were enriched in Lactobacillus, a bacterial genus with known probiotic and psychobiotic properties. Although further studies are needed to validate our findings, our work supports the intriguing opportunity that different behavioral phenotypes in dogs may be associated with peculiar gut microbiome layouts, suggesting possible connections between the gut microbiome and the central nervous system and indicating the possible adoption of probiotic interventions aimed at restoring a balanced host-symbiont interplay for mitigating behavioral disorders.

Spatio-temporal patterns of sarcoptic mange in red deer and Iberian ibex in a multi-host natural park.

The reliance on multiple hosts to survive is what makes the management and control of multi-host infectious agents challenging. Sarcoptes scabiei causes sarcoptic mange in a wide range of mammal species with ungulates being an important host. Little is known about the role different ungulates play in sustaining endemic transmission of the disease and no study has yet to describe the long-term multi-host sarcoptic infestation dynamics in free-ranging wildlife. Here, we explore 24 years of sarcoptic mange infestation data for two Mediterranean ungulate species, red deer and Iberian ibex, living in the Sierras de Cazorla, Segura y Las Villas Natural Park of southern Spain. The temporal analysis showed a clear seasonal pattern of infestation in both ungulates with a peak in early spring and a decline throughout the summer. The spatial analysis, however, showed that caprinae rather than cervidae is the most competent host for sarcoptic mange spreading and persistence. Considering that few studies have described the spatio-temporal pattern of mange outbreaks for long periods of time, the information reported in this work aims to improve our understanding of sarcoptic mange epizootic in wild ruminant populations.

Early colonisation and temporal dynamics of the gut microbial ecosystem in Standardbred foals.

BACKGROUND: Even if horses strictly depend on the gut microbiota for energy homeostasis, only a few molecular studies have focused on its characterisation and none on the perinatal gut microbial colonisation process. OBJECTIVES: To explore the perinatal colonisation process of the foal gut microbial ecosystem and the temporal dynamics of the ecosystem assembly during the first days of life. STUDY DESIGN: Longitudinal study. METHODS: Thirteen Standardbred mare-foal pairs were included in the study. For each pair, at delivery we collected the mare amniotic fluid, faeces and colostrum, and the foal meconium. Milk samples and faeces of both mare and foal were also taken longitudinally, until day 10 post-partum. Samples were analysed by means of next-generation sequencing of the 16S rRNA gene on Illumina MiSeq. RESULTS: Our findings suggest that microbial components derived from the mare symbiont communities establishes in the foal gut since fetal life. After birth, an external transmission route of mare microorganisms takes place. This involves a rapid and dynamic process of assembling the mature foal gut microbiome, in which the founder microbial species are derived from both the milk and the gut microbial ecosystems of the mare. MAIN LIMITATIONS: The inability to discriminate between live and dead cells, the possible presence of contaminating bacteria in low biomass samples (e.g. meconium and amniotic fluid), the limits of the phylogenetic assignment down to species level, and the presence of unassigned operational taxonomic units. CONCLUSIONS: Our data highlight the importance of mare microbiomes as a key factor for the establishment of the gut microbial ecosystem of the foal.

Risk factors of Leptospira infection in Mediterranean periurban micromammals.

Urbanization of natural areas can change abiotic factors, providing artificial sources of humidity in summer and decreasing variation of temperatures in winter. Our study aimed at document risk factors of infection in mammal reservoirs of pathogenic Leptospira in the human/wildlife interface of a large metropolitan area. We hypothesize that survival of Leptospira and thus their prevalence in animal reservoirs should be higher in residential areas than in natural habitats, especially after the hot, dry Mediterranean summers. We established the prevalence of Leptospira spp. and identified the serovars in 353 urine samples from micromammals (chiefly the wood mouse Apodemus sylvaticus, n = 266) using direct immunofluorescence and PCR. Animals were captured in spring and autumn, 2011-2012, in two natural parks and two adjacent residential areas in periurban Barcelona (NE Spain). Overall observed prevalence of infection was 11%, ranking between 8% and 13% in the better represented host species. We observed marked differences between seasons; the probability of finding a micromammal infected in spring was three times greater than in autumn (almost four times for wood mouse). Prevalence was not related with type of habitat, micromammal relative abundance or sex of the animal. Three Leptospira species were confirmed: Leptospira interrogans (47% of cases), Leptospira borgpetersenii (41%) and Leptospira kirschneri (12%). The serovars most commonly detected were those typically hosted by rodents, and serovars Ballum and Icterohemorrhagiae were the only ones found in autumn. People living in periurban Barcelona and those visiting the natural areas of the metropolitan area face hazard of infection with rodent-borne Leptospira, especially during spring.

Evaluation of Combination Drug Therapy for Treatment of Antibiotic-Resistant Inhalation Anthrax in a Murine Model.

Bacillus anthracis is considered a likely agent to be used as a bioweapon, and the use of a strain resistant to the first-line antimicrobial treatments is a concern. We determined treatment efficacies against a ciprofloxacin-resistant strain of B. anthracis (Cipr Ames) in a murine inhalational anthrax model. Ten groups of 46 BALB/c mice were exposed by inhalation to 7 to 35 times the 50% lethal dose (LD50) of B. anthracis Cipr Ames spores. Commencing at 36 h postexposure, groups were administered intraperitoneal doses of sterile water for injections (SWI) and ciprofloxacin alone (control groups), or ciprofloxacin combined with two antimicrobials, including meropenem-linezolid, meropenem-clindamycin, meropenem-rifampin, meropenem-doxycycline, penicillin-linezolid, penicillin-doxycycline, rifampin-linezolid, and rifampin-clindamycin, at appropriate dosing intervals (6 or 12 h) for the respective antibiotics. Ten mice per group were treated for 14 days and observed until day 28. The remaining animals were euthanized every 6 to 12 h, and blood, lungs, and spleens were collected for lethal factor (LF) and/or bacterial load determinations. All combination groups showed significant survival over the SWI and ciprofloxacin controls: meropenem-linezolid (P = 0.004), meropenem-clindamycin (P = 0.005), meropenem-rifampin (P = 0.012), meropenem-doxycycline (P = 0.032), penicillin-doxycycline (P = 0.012), penicillin-linezolid (P = 0.026), rifampin-linezolid (P = 0.001), and rifampin-clindamycin (P = 0.032). In controls, blood, lung, and spleen bacterial counts increased to terminal endpoints. In combination treatment groups, blood and spleen bacterial counts showed low/no colonies after 24-h treatments. The LF fell below the detection limits for all combination groups yet remained elevated in control groups. Combinations with linezolid had the greatest inhibitory effect on mean LF levels.

Episodic replacement of clotting factor concentrates does not prevent bleeding or musculoskeletal damage - the MUSFIH study.

PATIENTS AND METHODS: A longitudinal study was carried out in 255 children from 10 centres in nine developing countries over 5 years to assess the musculoskeletal outcome of children on episodic factor replacement. Outcome was documented by assessment of the annual joint bleeding rate (AJBR), WFH clinical and Pettersson radiological joint scores as well as the FISH score for activities. Of the 203 patients for whom data was available at the end of 5 years, 164 who had received only episodic treatment are included in this report. RESULTS: The median age at the beginning of the study was 10 years (IQR 7-12). The median clotting factor concentrate (CFC) usage was 662 IU kg-1 year-1 (IQ range: 280-1437). The median AJBR was 10 (IQ range: 5-17). The median AJBR was higher in the older children with the median being 5 for the 5 year old child, while it was 9 for the 10 year old and 11 for children older than 15. Given the episodic nature of the replacement therapy, those with a higher AJBR used significantly greater annual CFC doses (P < 0.001); The median change in WFH clinical score and Pettersson radiological score over the 5 years was 0.4/year for each, while the FISH deteriorated at a rate of 0.2/year with poor correlation of these changes with CFC dose. WFH and FISH scores were significantly worse in those with an AJBR of >3 per year (P = 0.001). The change in the Pettersson score was significantly more in those with an AJBR of >5 per year (P = 0.020). Significant changes in FISH scores were only noted after 10 years of age. CONCLUSION: Episodic CFC replacement over a large range of doses does not alter the natural course of bleeding in haemophilia or the musculoskeletal deterioration and should not be recommended as a long term option for treatment. Prophylaxis is the only way to preserve musculoskeletal function in haemophilia.

IgG subclasses of FVIII inhibitors in an argentine cohort of severe hemophilia A patients: Analysis by flow cytometry.

INTRODUCTION: FVIII inhibitors consist of a polyclonal population of antibodies. Previous studies have demonstrated different distribution of IgG subclasses. IgG4 was associated to high level of FVIII inhibitors and failure of immune tolerance induction (ITI) treatment. This study monitored the relative distribution of IgG subclasses of anti-FVIII in patients with severe hemophilia A (SHA). METHODS: Anti-FVIII antibodies were measured employing an immunomethod, developed in our laboratory, that combines flow cytometry (FC) with microspheres coupled (FVIII-m) or not (Control-m) to FVIII. Seventy-five patients with SHA were studied, 17 without inhibitors (Group I); 58 with inhibitor history, 13 low responders: (LR: Group II), and 45 high responders (HR: Group III). Eight patients undergoing ITI were also included. RESULTS: We found anti-FVIII antibodies in 11 of 27 patients (40%) without inhibitors and in 45 of 48 with inhibitors at the moment of the study. IgG4 was predominant only in the Group III: P=0.02 in patients with low level of inhibitors and P=0.0001 with high titer of inhibitors. Longitudinal analysis performed on patients undergoing ITI showed a gradual decrease of IgG4 values that was associated to improvement of clinical parameters during treatment. CONCLUSION: We suggest the use of the FC method to supplement functional traditional assays and to help to improve the management of patients with SHA.

Gut microbiota and hematopoietic stem cell transplantation: where do we stand?

Advances in biological techniques have potentiated great progresses in understanding the interaction between human beings and the ∼10 to 100 trillion microbes living in their gastrointestinal tract: gut microbiota (GM). In this review, we describe recent emerging data on the role of GM in hematopoietic stem cell transplantation, with a focus on immunomodulatory properties in the immune system recovery and the impact in the development of the main complications, as GvHD and infections.

Wide exposure to Coxiella burnetii in ruminant and feline species living in a natural environment: zoonoses in a human-livestock-wildlife interface.

Assessment of the role of wild and domestic hosts as potential reservoirs of misdiagnosed zoonoses, such as Q fever by Coxiella burnetii, is an important public health issue today both for wildlife conservation and management of disease in human-livestock-wildlife interface. This study used ELISA, an indirect antibody, to research (2003-2013) C. burnetii infection in seven free-living wild and domestic ruminant species and in European wildcats (Felis silvestris). The animals studied were 0 European wildcats, 21 Spanish ibex (Capra pyrenaica), 314 red deer (Cervus elaphus), 556 fallow deer (Dama dama), 211 European mouflon (Ovis aries musimon), eight roe deer (Capreolus capreolus), 407 bovines (Bos taurus) and 3739 sheep (Ovis aries). All the animals shared the same habitat in the Serranía de Cuenca Natural Park (Castile-La Mancha, Spain). The study area is an example of human-domestic-wildlife interface where people and domestic animals live in close proximity to wildlife. Observed C. burnetii seropositive frequencies were: 33·3% European wildcats, 23·8% Spanish ibex, 22·5% domestic sheep 1·5% red deer, 1·4% European mouflon, 0·24% cattle, 0·18% fallow deer and 0% roe deer. The study found a wide C. burnetii prevalence of previous and present exposure in wild and domestic ruminant hosts in the Serranía de Cuenca Natural Park and reports the first evidence of C. burnetii exposure in free-living European wildcats.

Treatment of subchondral cysts in patients with haemophilia.

INTRODUCTION: The haemophilic arthropathy is a disabling disease that causes chronic pain and functional limitation, due to recurrent intra-articular bleeding, with impaired quality of life. OBJECTIVE: The aim of this work is to present our 24-year experience in the treatment of subchondral cysts filled with hydroxyapatite coralline in patients with haemophilia. PATIENTS AND METHOD: Thirty-seven male patients with forty-nine cystic lesions were treated and evaluated between 1990 and 2014. Thirty four patients were haemophilia type A, three were haemophilia type B, two patients had inhibitors to factor VIII. The average age was 23.6 years. The average follow-up was 10 years. The lesions were located: twenty-four in the tibia (49%), six in the talus (12.2%), seven in the ulna (14.4%), five in the humerus (10.2%), five in the femur (10.2%) and two in the distal radius (4%). In all patients' radiographs, Computed Tomography and Magnetic Resonance Imaging were performed before surgery. The lesions were treated when the injury was greater than 15% of the joint area, and when the joint area was greater than 1 cm of diameter. Surgical technique consisted of aspirating the cyst contents and refilling it with hydroxyapatite coralline. RESULTS: In forty-eight cysts, restitution of bone structure was achieved by impaction of hydroxyapatite coralline. The average time of bone restoration was 10 months. Only one patient required a second intervention. CONCLUSION: The treatment of subchondral cyst in PWH by aspiration and filling with hydroxyapatite coralline allows bone restoration and delays deterioration of the joint treated.

Postoperative Autologous Reinfusion in Total Knee Replacement.

Surgeries for total knee replacement (TKR) are increasing and in this context there is a need to develop new protocols for management and use of blood transfusion therapy. Autologous blood reduces the need for allogeneic blood transfusion and the aim of the present study was to verify the safety and the clinical efficacy. An observational retrospective study has been conducted on 124 patients, undergoing cemented total knee prosthesis replacement. Observed population was stratified into two groups: the first group received reinfusion of autologous blood collected in the postoperative surgery and the second group did not receive autologous blood reinfusion. Analysis of data shows that patients undergoing autologous blood reinfusion received less homologous blood bags (10.6% versus 30%; p = 0.08) and reduced days of hospitalization (7.88 ± 0.7 days versus 8.96 ± 2.47 days for the control group; p = 0.03). Microbiological tests were negative in all postoperatively salvaged and reinfused units. Our results emphasize the effectiveness of this procedure and have the characteristics of simplicity, low cost (€97.53 versus €103.79; p < 0.01), and easy reproducibility. Use of autologous drainage system postoperatively is a procedure that allows reducing transfusion of homologous blood bags in patients undergoing TKR.

Epidemiology of fractures in patients with haemophilia.

INTRODUCTION: Haemophilia is an X-chromosome linked inherited bleeding disorder characterised by an anomaly synthesis of coagulation factor VIII (Haemophilia A) or factor IX (Haemophilia B). There is very little information on the magnitude and management of fractures in PWH in the literature regards the advance on replacement therapy. The purpose of this paper is to present our 28 years experience treating PWH who suffered fractures and evaluate the impact of access to treatment. MATERIALS AND METHODS: In the period 1986-2013, 151 fractures in 141 PWH were treated, 125 patients type A (88.7%), 12 type B (8.5%) and 4 (2.8%) with von Willebrand's disease. For the sake of analysis we divided the fractures in five groups: 1986-1990: 25, 1991-1995: 35, 1996-2001: 33, 2002-2007: 31, and 2008-2013: 27; and classified the fractures in lower limb (LL) and upper limb (UL). We also considered the age at which the fractures occurred. RESULTS: However the incidence of presentation of the fractures of the upper limb and lower limb changed through the years, being more frequent in the LL in the first period analysed (76% LL vs. 24% UL) and in the UL in the latter one (63% UL vs. 37% LL), the difference was statically significant (p=0.0168). In the relation with the age, the 1986-1990 cohort vs. 2008-2013 cohort reached statistical significance (p: 0.035). Regarding treatment, 121 fractures were treated in a non invasive way, the others 30 fractures, were treated with internal fixation. The patient treated with internal fixation had less mal-alignment, and delay consolidation. DISCUSSION: This is the largest series of fractures in PWH published. We show a higher incidence of LL fractures in the first period analysed (1986-1990), over time, the ratio LL/UL changed as UL fractures became more frequent. This change is due to the access of the treatment and specifically to the prophylaxis. CONCLUSION: Fractures in PWH have changed their pattern, becoming more common in the UL than in the LL, lowering the age at which they occur and being less frequent. We believe that the advent of new and accessibly treatments, decreased the development of orthopaedic complications and favours the improvement in quality of life of PWH.

Gut microbiota trajectory in pediatric patients undergoing hematopoietic SCT.

Acute GvHD (aGvHD) is the main complication of hematopoietic SCT (HSCT) during the treatment of hematological disorders. We carried out the first longitudinal study to follow the gut microbiota trajectory, from both the phylogenetic and functional points of view, in pediatric patients undergoing HSCT. Gut microbiota trajectories and short-chain fatty acid production profiles were followed starting from before HSCT and through the 3-4 months after transplant in children developing and not developing aGvHD. According to our findings, HSCT procedures temporarily cause a structural and functional disruption of the gut microbial ecosystem, describing a trajectory of recovery during the following 100 days. The onset of aGvHD is associated with specific gut microbiota signatures both along the course of gut microbiota reconstruction immediately after transplant and, most interestingly, prior to HSCT. Indeed, in pre-HSCT samples, non-aGvHD patients showed higher abundances of propionate-producing Bacteroidetes, highly adaptable microbiome mutualists that showed to persist during the HSCT-induced ecosystem disruption. Our data indicate that structure and temporal dynamics of the gut microbial ecosystem can be a relevant factor for the success of HSCT and opens the perspective to the manipulation of the pre-HSCT gut microbiota configuration to favor mutualistic persisters with immunomodulatory properties in the gut.

Haemophilia pseudotumours in patients with inhibitors.

Development of inhibitors against factor VIII (FVIII) or FIX is the most serious complication of replacement therapy in patients with haemophilia. Haemophilic pseudotumours in a patient with inhibitors can lead to devastating consequences. The aim of this study is to show our experience in the treatment of 10 pseudotumours in 7 patients with inhibitors who were treated by the same multidisciplinary team in the period between January 2000 and March 2013. Seven severe haemophilia A patients were treated at the Haemophilia Foundation in Buenos Aires, Argentina, for 10 pseudotumours. Eight were bone pseudotumours and two soft tissue. All patients underwent imaging studies at baseline to assess the size and content of the lesion. The patients received Buenos Aires protocol as conservative treatment of their pseudotumours for 6 weeks, after which they were evaluated. Only one patient responded to conservative treatment. Surgery was performed on the others six patients, since their pseudotumours did not shrink to less than half their original size. Any bleeding in the musculoskeletal system must be treated promptly in order to prevent pseudotumours. When pseudotumours do appear in inhibitor patients, they can be surgically removed when patients received proper haemostatic coverage, improving the quality of life of these patients.

Phenotype-genotype correlations in hemophilia A carriers are consistent with the binary role of the phase between F8 and X-chromosome inactivation.

BACKGROUND: The recessive X-linked disorder hemophilia A (HA) is rarely expressed in female carriers, most of whom express about half of normal factor VIII activity ( FVIII: C). OBJECTIVE: To propose an integrative assessment model for the binary role of the phase between the mutated F8 and the active X-chromosome (Xa) in FVIII: C in HA carriers. METHODS: We studied 67 females at risk of severe HA, comprising five symptomatic females ( FVIII: C < 1.5 IU dL(-1) ) and 14 controls. A correlation study between FVIII: C (observed vs. expected) and X-chromosome inactivation (XCI) patterns (XIPs; androgen receptor gene [AR] system) in blood leukocyte DNA was performed in carriers, by comparison of a model correlating FVIII: C and XIP with arbitrary models devoid of biological significance, and with FVIII: C levels in non-carriers (mean model) as a proxy from background data dispersion not influenced by XIP. RESULTS: We provide proof-of-concept example from a family presenting with extremely skewed XIPs in which the severe HA phenotype appeared in a heterozygous carrier of a crossover between AR and F8 loci that phased the mutated F8 with the maternally inherited Xa. Furthermore, four cases of severe HA affected women who had a combination of a heterozygous F8 mutation and extremely skewed XIPs in leukocytes or oral mucosa are presented. Correlation analyses between FVIII: C levels and XIPs in carriers (n = 38) but not in non-carriers (n = 20) showed highly significant differences between the proposed correlation model and models without biological significance. The data support a binary influence of XCI, either increasing or decreasing the FVIII: C, subject to the underlying phase set between the F8 mutation and XCI. CONCLUSIONS: Our evidence suggests that the phase between XCI and mutated F8 acts as a molecular switch conditioning FVIII: C levels and HA expression in carriers.

Musculoskeletal evaluation in severe haemophilia A patients from Latin America.

There is a paucity of literature on haemophilia treatment in Latin American countries, a region characterized by rapidly improving systems of care, but with substantial disparities in treatment between countries. The aim of this study was to evaluate the musculoskeletal status of haemophilia patients from Latin America and to examine the relationship between musculoskeletal status and treatment practices across countries. The Committee of Latin America on the Therapeutics of Inhibitor Groups conducted a survey of its member country representatives on key aspects of haemophilia treatment in 10 countries. Musculoskeletal status of patients was obtained during routine comprehensive evaluations between March 2009 and March 2011. Eligible patients had severe haemophilia A (factor VIII <1%) without inhibitors (<0.6 BU mL(-1) ) and were ≥5 years of age. Musculoskeletal status was compared between three groups of countries, based primarily on differences in the availability of long-term prophylaxis. Overall, 143 patients (5-66 years of age) were enrolled from nine countries. In countries where long-term prophylaxis had been available for at least 10 years (Group A), patients aged 5-10 years had significantly better mean World Federation of Hemophilia clinical scores, fewer target joints and fewer affected joints than patients from countries where long-term prophylaxis has been available for about 5 years (Group B) or was not available (Group C). In Latin America, the musculoskeletal status of patients with severe haemophilia without inhibitors has improved significantly in association with the provision of long-term prophylaxis. As more countries in Latin America institute this practice, further improvements are anticipated.