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Cutaneous lesions described as chilblain lupus occur in the context of familial chilblain lupus or Aicardi-Goutières syndrome. To date, seven genes related to Aicardi-Goutières syndrome have been described. The most recently described encodes the cytosolic double-stranded RNA receptor IFIH1 (also known as MDA5), a key component of the antiviral type I interferon-mediated innate immune response. Enhanced type I interferon signalling secondary to gain-of-function mutations in IFIH1 can result in a range of neuroinflammatory phenotypes including classical Aicardi-Goutières syndrome. It is of note that none of the patients with a neurological phenotype so far described with mutations in this gene was reported to demonstrate cutaneous involvement. We present a family segregating a heterozygous pathogenic mutation in IFIH1 showing dermatological involvement as a prominent feature, variably associated with neurological disturbance and premature tooth loss. All three affected individuals exhibited increased expression of interferon-stimulated genes in whole blood, and the mutant protein resulted in enhanced interferon signalling in vitro, both in the basal state and following ligand stimulation. Our results further extend the phenotypic spectrum associated with mutations in IFIH1, indicating that the disease can be confined predominantly to the skin, while also highlighting phenotypic overlap with both Aicardi-Goutières syndrome and Singleton-Merten syndrome., (© 2015 British Association of Dermatologists.)

The type I interferon system is integral to human antiviral immunity. However, inappropriate stimulation or defective negative regulation of this system can lead to inflammatory disease. We sought to determine the molecular basis of genetically uncharacterized cases of the type I interferonopathy Aicardi-Goutières syndrome and of other undefined neurological and immunological phenotypes also demonstrating an upregulated type I interferon response. We found that heterozygous mutations in the cytosolic double-stranded RNA receptor gene IFIH1 (also called MDA5) cause a spectrum of neuroimmunological features consistently associated with an enhanced interferon state. Cellular and biochemical assays indicate that these mutations confer gain of function such that mutant IFIH1 binds RNA more avidly, leading to increased baseline and ligand-induced interferon signaling. Our results demonstrate that aberrant sensing of nucleic acids can cause immune upregulation.

Aicardi-Goutières syndrome is an inflammatory disorder resulting from mutations in TREX1, RNASEH2A/2B/2C, SAMHD1, or ADAR1. Here, we provide molecular, biochemical, and cellular evidence for the pathogenicity of two synonymous variants in RNASEH2A. Firstly, the c.69G>A (p.Val23Val) mutation causes the formation of a splice donor site within exon 1, resulting in an out of frame deletion at the end of exon 1, leading to reduced RNase H2 protein levels. The second mutation, c.75C>T (p.Arg25Arg), also introduces a splice donor site within exon 1, and the internal deletion of 18 amino acids. The truncated protein still forms a heterotrimeric RNase H2 complex, but lacks catalytic activity. However, as a likely result of leaky splicing, a small amount of full-length active protein is apparently produced in an individual homozygous for this mutation. Recognition of the disease causing status of these variants allows for diagnostic testing in relevant families., (© 2013 WILEY PERIODICALS, INC.)

Coats plus is a highly pleiotropic disorder particularly affecting the eye, brain, bone and gastrointestinal tract. Here, we show that Coats plus results from mutations in CTC1, encoding conserved telomere maintenance component 1, a member of the mammalian homolog of the yeast heterotrimeric CST telomeric capping complex. Consistent with the observation of shortened telomeres in an Arabidopsis CTC1 mutant and the phenotypic overlap of Coats plus with the telomeric maintenance disorders comprising dyskeratosis congenita, we observed shortened telomeres in three individuals with Coats plus and an increase in spontaneous γH2AX-positive cells in cell lines derived from two affected individuals. CTC1 is also a subunit of the α-accessory factor (AAF) complex, stimulating the activity of DNA polymerase-α primase, the only enzyme known to initiate DNA replication in eukaryotic cells. Thus, CTC1 may have a function in DNA metabolism that is necessary for but not specific to telomeric integrity.

There is considerable concern about pollution of surface waters with P. Although most of the research has focused on inorganic P in surface runoff, it has recently become possible to easily follow the fate of soluble organic P forms in soils and waters. Two experiments were performed to compare the relative mobility and soil fixation affinity of orthophosphate monoesters, orthophosphate diesters, and soluble inorganic P. We used three P substrates, 4-methylumbelliferyl phosphate (MUP), deoxyribonucleic acid (DNA), and KH(2)PO(4) in (i) a soil column experiment and (ii) a soil P adsorption test tube experiment. Shortly after columns were prepared, approximately two pore volumes of 0.005 M CaCl(2) were passed through 25 cm length columns containing 10 cm of loamy sand amended with approximately 10 mg P as MUP, DNA, or KH(2)PO(4) above 15 cm of nonamended loamy sand. The total net quantity of 757.8 microg P 2L(-1) of orthophosphate diesters in the leachate from the DNA columns exceeded the net quantity of orthophosphate monoesters in leachate from the MUP columns (4.6 microg P 2L(-1)) and soluble inorganic P from the KH(2)PO(4) columns (34.0 microg P 2L(-1)). Adsorption of soluble organic and inorganic P in the test tube experiment yielded similar results: DNA, containing orthophosphate diesters, had a relatively low affinity for soils. In both experiments, high concentrations of other P compounds were identified in samples treated with organic P substrates, suggesting enzymatic hydrolysis by native soil phosphatase enzymes. These findings indicate that repeated application of organic forms of P could lead to significant leaching of P to ground water.

Unwanted microbial interference in samples used for biological assays of P availability has routinely been eliminated by autoclaving samples before inoculation with algae. Twenty-three soils were selected to evaluate the relationship between algal growth in P-deficient solutions containing small quantities of soil and the level of P determined by a variety of tests used to evaluate P availability in soils and sediments. Soils were either autoclaved or not before addition to flasks containing P-starved algae in a nutrient solution without P. Compared to non-autoclaved samples, autoclaving soil resulted in approximately 60% more available P as estimated by increased algal growth. However, algal growth in the presence of autoclaved soil was highly correlated with growth in the presence of non-autoclaved samples. There was no consistent change in the correlations (r) between autoclaving or non-autoclaving samples in the relationships of algal numbers with P extracted by a number of soil tests. The effect of autoclaving soil on soluble P was also evaluated for a subset of six soils. Autoclaved soils had significantly greater concentrations of soluble P than non-autoclaved soils, with 78% more orthophosphate monoesters, 60% more orthophosphate diesters, and 54% more soluble inorganic P. Inhibition of algal growth may have occurred with two high-Zn soils that produced relatively low numbers of algae despite being very high in estimated available P by all extraction methods. Removing those samples from the calculations dramatically improved correlations between soil P measured by various methods and algal growth. With these two soils removed from calculations, algal growth with autoclaved soil was most highly correlated with Olsen P (r = 0.95), with other correlations as follows: Fe-oxide strip (r = 0.80), Mehlich 3 (r = 0.75,), modified Morgan (r = 0.61), and Bray-Kurtz 1 (r = 0.57).

Aim: To determine if the superficial digital flexor tendon (SDFT) of young Thoroughbred horses changed in size and echogenicity in association with early race training., Methods: Cross-sectional area (CSA) and echogenicity were determined ultrasonographically at five levels of the SDFT of the forelimbs of 2-year-old fillies (n=14), corresponding to 4, 8, 12, 16 and 20 cm distal to the accessory carpal bone (DACB). Measurements were made before and after a 13-week period in which a trained group of seven horses was compared with another group of seven untrained horses., Results: Level below the accessory carpal bone had a significant effect on CSA and Level 8 was smaller than all other levels except Level 12, while Level 12 was smaller than Levels 4 and 20 but not different from Levels 8 and 16. There was a significant interaction between level and time due to effects observed at Level 8. The CSA at Level 8 measured pre-training was different from that of Levels 4 and 20 in both pre- and post-training groups (p<0.05), but when measured post-training was not different from any other measurement. There was also a significant interaction between treatment group and time. There was no difference between CSA for the untrained and trained groups at the pre-training observation (p=0.9), but post-training the CSA (pooled over all levels) in trained horses was significantly larger than that of the untrained horses both post-training (p=0.019) and pre-training (p=0.034), and was not different from the pre-training CSA recorded in the trained group (p=0.29). Treatment group had no effect on echogenicity (p=0.43), while echogenicity was less at the end of the trial in both trained and untrained horses (p<0.001)., Conclusions: Early training for racing was associated with an increase in mean CSA of the SDFT. Other factors such as age and maturity may play a role in limiting this increase.

A number of methods for the local delivery of drugs to musculoskeletal tissues in the horse are now available. Further research is required to document the disposition of drugs delivered by such methods and to correlate this information with efficacy. Perhaps the greatest potential area for the methods discussed is the treatment of synovial and bone infections. To be able to provide high and sustained therapeutic concentrations of antimicrobials to the site of infection should increase the chances of success in such cases. These methods of drug delivery need to be used in conjunction with other management procedures, however, including bacterial culture and sensitivity procedures, systemic antimicrobials, surgical drainage, removal of dead bone or surgical implants, establishment of fracture stability, use of autogenous bone grafts, systemic NSAIDs, and rest.

Immunosuppression as a consequence of acute and chronic stress can increase the susceptibility of cattle to a range of infectious diseases. In order to develop a panel of immune function assays for investigating the effects of potential stressors on immune competence in cattle, the effect of treatment with short- and long-acting preparations of the synthetic glucocorticoid dexamethasone was examined. Short-acting dexamethasone (dexamethasone sodium phosphate 0.08 mg/kg) followed 37 h later by long-acting dexamethasone (dexamethasone-21 isonicotinate 0.25 mg/kg) was injected intramuscularly and blood was collected to assess immune functions at intervals over the subsequent 11 days from 6 treated and 6 control Hereford steers. Dexamethasone induced leukocytosis (neutrophilia, eosinopenia, lymphopenia, monocytosis), an increased neutrophil:lymphocyte ratio, an elevated percentage of CD4+ lymphocytes, a decreased total CD8+ lymphocyte count, decreased total and percentage WC1+ lymphocytes, an elevated percentage of IL-2 receptor alpha (IL-2Ralpha)+ lymphocytes, and an elevated percentage of B lymphocytes. In vitro chemotaxis of peripheral blood neutrophils to human C5a and ovine IL-8 was increased by dexamethasone treatment. Lymphocyte proliferation in the presence of phytohaemagglutinin, and serum concentrations of IgM, but not IgA or IgG1, were suppressed by dexamethasone treatment, whereas mitogen-induced production of interferon-gamma (IFN-gamma), neutrophil expression of CD18, neutrophil myeloperoxidase activity and natural killer (NK) cell activity were not influenced by dexamethasone treatment. The results indicate the potential for haematology and immune function assays to reflect elevated activity of the hypothalamic-pituitary-adrenocortical axis in cattle. Immunological parameters may thus provide a useful adjunct to cortisol and behavioural observations for assessing the impact of stress on the welfare of cattle.

One of two groups of sheep was immunosuppressed with the glucocorticoid, dexamethasone, at the time of the first but not of the second of two booster vaccinations with tetanus toxoid given at an interval of 28 days. Treatment with dexamethasone decisively reduced the anti-tetanus antibody response to the first booster vaccination and affected both IgM and IgG1 antibody. However, antibody titres increased after the second booster vaccination in the treated sheep and were similar in size to those in the untreated sheep which rose in stepwise fashion after each booster vaccination. The differences in response imply that processes involved in displaying an anamnestic response and recalling previously established memory are sensitive to glucocorticoids. Accordingly, they can be separated from the glucocorticoid-resistant processes that lead to the expansion of immunological memory following multiple exposures to an antigen.

Neurofilament accumulations are characteristic of a number of neurological conditions including amyotrophic lateral sclerosis, giant axonal neuropathies and several chemically-induced neuropathies. Although the mechanism(s) leading to neurofilament accumulation are unknown, it is possible that similar processes occur both in disease and in chemically-induced neuropathies. Understanding the mechanism(s) of chemically-induced neurofilament accumulation, which is more amenable to experimental manipulation, may give insight into the neurological diseases they mimic. We have compared the effects of two chemically-dissimilar neurotoxins, 2,5-hexanedione and acrylamide, on neurofilaments in the human neuroblastoma cell line, SH-SY5Y. Both undifferentiated and differentiated SH-SY5Y cells were exposed to 2,5-hexanedione or acrylamide and changes in cytoskeletal organization examined by immunofluorescence and electron microscopy. Although distinct morphological differences have previously been characterized in the neuropathies induced by 2,5-hexanedione and acrylamide in vivo, we have found that both compounds had similar direct effects on neurofilaments in SH-SY5Y cells, inducing formation of perikaryal inclusion bodies. In addition, differentiated SH-SY5Y cells were more sensitive to both 2,5-hexanedione and acrylamide compared with undifferentiated cells. These similar effects of 2,5-hexanedione and acrylamide lend further support that a common mechanism(s) may lead to neurofilament accumulation in these neuropathies. SH-SY5Y cells provide a useful model to investigate further the biochemical basis of neurofilament accumulation.

An endoscopic survey of young race horses was performed to examine the prevalence and character of laryngeal movements during quiet respiration. The main aim was to determine whether those arytenoid movements which could possibly reflect the efficiency of left dorsal cricoarytenoid muscle function changed over a period of time. Of the 452 horses examined, 439 were Thoroughbreds and 23 were Standardbreds, 250 were less than 2 years of age (6-21 months), and 202 were 2 years old. One hundred and nine of these horses were examined again 16 months later. Arytenoid movements were given one of four grades. Grades 1 and 2 were considered normal and unlikely to be the result of abnormal left dorsal cricoarytenoid muscle function, whilst grades 3 and 4 were considered likely, or almost certainly, the result of abnormal left dorsal cricoarytenoid muscle function. The percutaneous prominence of the muscular process of left and right arytenoid cartilages, endoscopic arytenoid movement on left and right sides, age, sex and breed was recorded. Chi squared analysis was used to determine the association between age, breed, sex and the other recorded variables, and the presence or absence of abnormal laryngeal movements. At the first examination, 48% of the horses had grade 1, 37% grade 2, 15% grade 3 and 0.2% grade 4 left laryngeal movements. Of the horses examined I6 months later, 52% had grade 1, 33% grade 2, 14% grade 3 and 1% grade 4 left laryngeal movements. Fifteen percent of horses with grade 1 and 9% with grade 2 initially were found to be grade 3 at the subsequent examination. Conversely, 53% of horses with grade 3 initially were found to be grade 1 and 21% grade 2 at the subsequent examination. One horse that was grade 3 at the initial examination was grade 4 at the subsequent examination. Overall, 43% of horses were graded the same, 29% were given a better grade and 28% were given a worse grade. Age and sex were not associated with abnormal left laryngeal movements. The presence of abnormal arytenoid movements was significantly less in Standardbreds, but significantly higher in those horses that had a more prominent muscular process of the left arytenoid cartilage. The number of grade 2 and 3 laryngeal movements recorded on the left side was significantly higher than the right. It was concluded that asymmetrical laryngeal movements are common in young race horses; at this age laryngeal movements may interchange between what is considered normal and abnormal; the proportion of young horses with normal or minor variations in their left arytenoid movements that develop more obvious degrees of asynchrony is low (12%); and the proportion of horses considered to have endoscopic evidence of deficient left abductor muscle function that eventually develop laryngeal hemiplegia is also low (5%).

Objective: To describe clinical findings, ultrasonographic features and outcome of injury to the branches of insertion of the superficial digital flexor (SDF) tendon in horses., Design: Retrospective study of 14 cases., Procedure: Fourteen Thoroughbred horses with tendonitis affecting the branches of insertion of the SDF tendon were examined for lameness, location and amount of swelling, and the presence of other musculoskeletal abnormalities. The flexor tendons were assessed by ultrasonographic examination, and recommendations were made for management of the cases. Outcome was assessed by re-examination of some horses, direct communication with the owner or trainer, and examination of race records., Results: The lateral SDF branch was affected in 10 horses; the medial branch in three, and both branches in one horse. Two horses had concurrent injuries to the SDF tendon in the metacarpal region of the contralateral limb. Ultrasonographic findings included swelling of the affected SDF branch, peritendinous fluid accumulation, disruption of normal fibre alignment on sagittal scan, and variable loss of echogenicity. As healing occurred, there was return of normal echogenicity, but normal fibre alignment did not return completely and apparent adhesions formed between the affected SDF branch and adjacent structures. Seven of 10 horses which returned to their previous use were able to compete without further tendon injury. Recurrence of injury occurred in one case, and another two horses developed tendonitis in the metacarpal region. One horse was retired from racing but was able to compete at dressage without recurrence of injury. Two horses were retired for breeding without returning to training, and one horse was sold and lost to follow up but did not race., Conclusion: The prognosis is fair for return to previous use following injury to the branches of insertion of the SDF tendon in athletic horses.

Successful treatment of a comminuted frontal plane fracture of the distal phalanx in a horse is described. The bone fractured through the solar canal, close to the insertion of the deep digital flexor tendon. A hoof case was used to reduce bending and tensile stresses on the solar surface by limiting expansion of the hoof wall during weightbearing. In addition, the heel was elevated, using 3 degrees wedge pads incorporated within the hoof cast, to reduce distraction at the fracture site caused by the pull of the deep digital flexor tendon. Two casts were used over a 4-month period. Complete radiographic healing of the fracture was evident 4 months after injury. At this time the horse was lame at the trot. The owner reported the horse to be sound 7 months after injury. Reexamination 12 months after injury revealed a clinically normal foot and no lameness.

Exploratory laparotomy of an adult dairy cow, examined because of acute signs of persistent abdominal pain, revealed a firm pulsatile mass with associated fremitus just distal to the origin of the cranial mesenteric artery. The cow died acutely 2.5 days after surgery. A dilated, thin-walled, sacculated aneurysm, which had ruptured, was located along the proximal portion of the cranial mesenteric artery. It was postulated that the aneurysm developed secondary to structural defects in the arterial wall, but caused no clinical signs until enlargement and local tissue stretching or circulatory disturbances caused intestinal ischemia, resulting in abdominal pain. Aneurysms of visceral arteries in cattle should be considered as another differential diagnosis for signs of abdominal pain after more common causes such as severe bloat, mesenteric root volvulus, intussusception, cecal dilatation/volvulus, and uterine torsion have been excluded.

Plasma (P), synovial fluid (SF) and lymph (L) concentrations of gentamicin were studied in two trials. A lymph vessel in the hindlimb was cannulated. The day after surgery (trial A), P and L samples were collected for 12 h after intravenous injection of gentamicin sulphate at 2.2 mg/kg dose rate. Approximately 48 h after surgery (trial B), the fetlock joint of the cannulated hindlimb was catheterized and P, SF and L samples collected for 12 h after a similar intravenous injection. The kinetic parameters were similar to those in other reports and did not differ between trials (P < 0.05). The P, L and SF disposition profiles were similar. The 95% confidence interval for P & L concentrations overlapped 2-3 h after injection. Thereafter, parallelism between L and P concentrations was observed, but L concentrations were on average 60% higher than P concentrations, and elimination from L was slower than from P. The mean L/SF and P/SF ratios were 1.54 +/- 0.2 and 1.25 +/- 0.2, 2-4 h after injection. Gentamicin elimination from SF appeared to be slower than from L and P. Lymph cannulation is a viable technique for antibiotic disposition studies. A sample of any of the fluids 3 h after injection was representative of the others. While SF concentrations were of limited value for predicting tissue fluid (L) concentrations 3-8 after injection, P concentrations were a useful index.

Lambs with genetically determined increased immunological responsiveness to Trichostrongylus colubriformis (high responders) had more eosinophils in cutaneous reactions to the mitogen phytohaemagglutinin (PHA) both before and during infection compared with those bred for susceptibility (low responders). In contrast, eosinophil numbers in both blood and cutaneous reactions elicited by third-stage T. colubriformis larval antigen were similar in high and low responders before infection. Following vaccination and challenge, high responders had elevated eosinophil numbers in blood and antigen-stimulated skin. In unselected sheep, although eosinophil numbers in cutaneous reactions to PHA were related to responsiveness to a challenge infection with T. colubriformis, there was a closer relationship between blood eosinophil numbers and responsiveness. Infection with T. colubriformis increased eosinophil numbers in cutaneous reactions to PHA and appeared to augment the difference between eosinophil counts in high and low responder sheep. Measurement of the ability to produce eosinophil activating factors, or for eosinophils to respond to such factors might therefore be useful in identifying individual sheep with increased responsiveness to T. colubriformis infection.

Medical professionals are ethically obligated to document the quality of the care they provide for patients. Society's demands and patients' expectations make quality assurance an issue of concern to all health care providers. The Joint Commission on Accreditation of Healthcare Organizations has formulated a detailed process for monitoring clinical activities and evaluating quality. This report reviews the rationale for and recommended approach to monitoring and evaluating quality assurance.

Cross-protective immunity between the nematode parasites, Haemonchus contortus and Trichostrongylus colubriformis, was examined in sheep vaccinated with irradiated larvae of either species. Secondary immunological responsiveness stimulated in this manner protected only against challenge infection with the species used for vaccination. Significant cross-protective immunity was not observed. Titres of serum antibody to an extract of adult but not infective larval T. colubriformis reflected the specificity for protective immunity. Immediate hypersensitivity skin reactions to nematode extracts did not reflect the antigen-specificity for protective immunity.