TF protein of Sindbis virus antagonizes host type I interferon responses in a palmitoylation-dependent manner.

Sindbis virus (SINV) produces the small membrane protein TF from the 6K gene via a (-1) programmed ribosomal frameshifting. While several groups have shown that TF-deficient virus exhibits reduced virulence, the mechanism(s) by which this occurs remain unknown. Here, we demonstrate a role for TF in antagonizing the host interferon response. Using wild-type and type 1 interferon receptor-deficient mice and primary cells derived from these animals, we show that TF controls the induction of the host interferon response at early times during infection. Loss of TF production leads to elevated interferon and a concurrent reduction in viral loads with a loss of pathogenicity. Palmitoylation of TF has been shown to be important for particle assembly and morphology. We find that palmitoylation of TF also contributes to the ability of TF to antagonize host interferon responses as dysregulated palmitoylation of TF reduces virulence in a manner similar to loss of TF.

Identification of complications in paediatric cerebral palsy treated with intrathecal baclofen pump: a descriptive analysis of 15 years at one institution.

PURPOSE: Intrathecal baclofen (ITB) treatment is used with increasing frequency in the cerebral palsy population. We describe the complications of ITB treatment, the incidence of complications, and our experience with their treatment. METHODS: In a period of 15 years, 341 paediatric patients with cerebral palsy treated with ITB were evaluated. Device problems associated with the catheter or pump, or infection and complications such as cerebrospinal fluid (CSF) leak and postdural spinal headache, were reviewed. Infection was classified as early (≤ 90 days) or late (> 90 days) according to the time of onset. RESULTS: The infection rate was 6.9% per procedure (50/720) and 14.6% per patient (50/341) over a mean 6.3 ± 3.9 years. There was a positive correlation between the risk of infection and preoperative comorbidities including epilepsy/seizure history, feeding tube, and mixed type cerebral palsy (p < 0.05, p = 0.03, p = 0.01, respectively). Eighty-five (24.9%) patients experienced 90 CSF leak episodes; 61 of these 85 patients had headache complaints as a result of CSF leak. There was a positive correlation between the risk of early infection and CSF leak (p < 0.05). CONCLUSIONS: The most common complication related to ITB was associated with pump and catheter problems. The rate of complications with the use of ITB is relatively high; however, based on the literature reports, it is the most effective treatment for severe spasticity and dystonia in patients with severe cerebral palsy. LEVEL OF EVIDENCE: III.

Foot and ankle deformities in children with Down syndrome.

PURPOSE: Foot and ankle deformities are common orthopaedic disorders in children with Down syndrome. However, radiographic measurements of the foot and ankle have not been previously reported. The aim of this study is to describe the foot and ankle deformity in children with Down syndrome. METHODS: Children who had foot and ankle radiographs in the standing weight-bearing position were selected. Three groups of patients were identified. The relationship of radiographic measurements with age, body mass index and pain is discussed. In all, 41 children (79 feet) had foot radiographs and 60 children (117 ankles) had ankle radiographs, with 15 children overlapping between Groups I and II. RESULTS: In Group I, hallux valgus deformity was seen before ten years of age and hallux valgus angle increased afterwards. Metatarsus adductus angle showed a significant increase (p = 0.006) with obesity and was higher in patients who had foot pain (p = 0.05). In Group II, none of the ankle measurements showed a significant difference with age or body mass index percentiles. Tibiotalar angle (TTA) and medial distal tibial angle (MDTA) were higher in patients who had ankle pain. In Group III, correlation analysis was performed between the different measurements with the strongest correlations found between TTA and MDTA. CONCLUSION: In children with Down syndrome, radiographic evaluation of the foot and ankle reveals higher prevalence of deformities than clinical examination. However, foot and ankle radiographs are needed only for symptomatic children with pain and gait changes. LEVEL OF EVIDENCE: Level IV - Prognostic Study.

Prophylactic antibiotics in soft-tissue procedures in children with cerebral palsy.

PURPOSE: Data on the benefits of perioperative prophylactic antibiotics in the paediatric population are lacking. In this study, we aimed to assess the rate of infection after isolated soft-tissue procedures in patients with cerebral palsy with and without preoperative prophylactic antibiotics between 2009 and 2015. METHODS: We reviewed the records of all children with cerebral palsy who underwent isolated soft-tissue procedures (on the upper and lower limb) at our hospital between 2009 and 2015. Children with at least 30-day postoperative follow-up were included. Children who had groin incisions were excluded. RESULTS: Two groups were identified: the antibiotic group (77 children with 203 incisions and 343 procedures) had one surgical site infection; the no-antibiotic group (48 children with 102 incisions and 177 procedures) had no surgical site infections. CONCLUSION: These results suggest that the use of preoperative antibiotics does not change the rate of postoperative surgical site infections. LEVEL OF EVIDENCE: Level III therapeutic study.

Arthroscopic knee anatomy in young achondroplasia patients.

PURPOSE: Achondroplasia is the most common form of skeletal dysplasia, affecting more than 250 000 individuals worldwide. In these patients, the developing knee undergoes multiple anatomical changes. The purpose of this study was to characterise the intra-articular knee anatomy in children with achondroplasia who underwent knee arthroscopy. METHODS: Records of achondroplasia patients who underwent knee arthroscopy between 2009 and 2014 were reviewed. Demographic data, operative reports, follow-up notes, MRI and arthroscopy images were reviewed. Bony, cartilaginous and ligamentous changes were noted. The trochlea sulcus angle was measured from intra-operative arthroscopic images. RESULTS: A total of 12 knee arthroscopies in nine patients were performed. The mean age at surgery was 16.9 years (12 to 22). In all patients, the indication for surgery was knee pain and/or mechanical symptoms that were refractory to non-operative treatment. Three anatomical variations involving the distal femur were found in all knees: a deep femoral trochlea; a high A-shaped intercondylar notch; and a vertically oriented anterior cruciate ligament. The average trochlea sulcus angle measured 123°. Pathology included: synovial plica (one knee); chondral lesions (three knees); discoid lateral meniscus (11 knees); and meniscal tears (six knees). All patients were pain-free and returned to normal activity at final follow-up. CONCLUSION: Children with achondroplasia have characteristic distal femur anatomy noted during knee arthroscopy. These variations should be considered normal during knee arthroscopy in these patients. Arthroscopic findings confirmed previous MRI findings within this specific population with the addition of a deep trochlear groove which was not previously reported.

Re-alignment and intramedullary rodding of the humerus and forearm in children with osteogenesis imperfecta: revision rate and effect on fracture rate.

PURPOSE: Intramedullary rodding is indicated for patients with osteogenesis imperfecta (OI) to manage deformity and help treat recurrent fractures. Historically, the focus of intramedullary stabilisation has been the lower extremity. Here we report our experience of intramedullary rodding of the humerus and forearm in children with OI and its impact on the fracture rate of those bone segments. PATIENTS AND METHODS: This is a retrospective chart review of all OI patients who have undergone re-alignment and intramedullary rodding of the humerus or forearm between October 1994 and February 2016. Patient demographics, surgical information, complications and pre-operative and post-operative fracture rates were gathered. RESULTS: A total of 45 upper extremity segments (26 humeri, 19 forearms) were rodded at an average age of 8.7 years (3.1 to 19.2). Of these, 15 (33.3%) of the bone segments required a return to the operating room at a mean 30.8 months (1 to 90) post-operatively. Fracture data was available for 24 of the bone segments. The average number of pre-operative and post-operative fractures was 3.58 (SD 2.84) and 0.46 (SD 0.72) respectively. The average pre-operative and post-operative fracture rates were 0.87 fractures/year (SD 0.47) and 0.10 fractures/year (SD 0.16) respectively. CONCLUSION: In this OI population, re-alignment and rodding appeared to reduce the fracture rate of the humerus and forearm. Among our population, one third returned to the operating room and one fifth required revision to a new intramedullary implant. This data may help families better understand the potential outcomes of upper extremity realignment and rodding and its effect on the rate of upper extremity fractures.

Long-term outcome of reconstruction of the hip in young children with cerebral palsy.

We reviewed the long-term radiological outcome, complications and revision operations in 19 children with quadriplegic cerebral palsy and hip dysplasia who underwent combined peri-iliac osteotomy and femoral varus derotation osteotomy. They had a mean age of 7.5 years (1.6 to 10.9) and comprised 22 hip dislocations and subluxations. We also studied the outcome for the contralateral hip. At a mean follow-up of 11.7 years (10 to 15.1) the Melbourne cerebral palsy (CP) hip classification was grade 2 in 16 hips, grade 3 in five, and grade 5 in one. There were five complications seen in four hips (21%, four patients), including one dislocation, one subluxation, one coxa vara with adduction deformity, one subtrochanteric fracture and one infection. A recurrent soft-tissue contracture occurred in five hips and ten required revision surgery. In pre-adolescent children with quadriplegic cerebral palsy good long-term outcomes can be achieved after reconstruction of the hip; regular follow-up is required.

Automated external defibrillator use at NCAA Division II and III universities.

OBJECTIVE: The placement of automated external defibrillators (AEDs) at collegiate sporting venues is a growing trend. The purpose of this study was to investigate the prevalence, location and past utilisation of AEDs at National Collegiate Athletic Association (NCAA) Division II and III universities. DESIGN: Cross-sectional survey. SETTING: NCAA Division II and III universities. PARTICIPANTS: Questionnaires were mailed to the head athletic trainer at NCAA Division II and III (N=711) colleges and universities in the fall of 2003. Findings were compared to previously published results at Division I institutions. MAIN OUTCOME MEASURE: Prevalence, location and past utilisation of AEDs. RESULTS: Completed surveys were returned by 254 NCAA Division II and III institutions for a 35.7% response rate (254/711). 205 (81%) institutions had at least one AED in the university athletic setting, with a median of 2 AEDs per institution (range 1-9). Athletic training rooms (75%) were the most likely location to place an AED. Twelve cases of AED use for sudden cardiac arrest were reported with 67% (8/12) occurring in older non-students, 16% (2/12) in intercollegiate athletes and 16% (2/12) in students (non-intercollegiate athletes). The AED deployed a shock in eight cases. 8 of 12 (66%) victims were immediately resuscitated, but only 4 survived to hospital discharge (overall survival 33%). None of the intercollegiate athletes or students survived. CONCLUSIONS: Most NCAA Division II and III institutions that responded to the survey have implemented AEDs in their athletic programs, although they have a lower prevalence of AEDs than previously reported at Division I universities. Although no benefit was demonstrated in a small number of intercollegiate athletes, AEDs were successfully used in older individuals on campus with cardiac arrest.

A high-capacity LC/MS system for the bioanalysis of samples generated from plate-based metabolic screening.

HPLC/MS is a linear technique characterized by serial injection and analysis of individual samples. Parallel-format high-throughput screens for druglike properties present a significant analytical challenge. Analysis speed and system ruggedness are key requirements for bioanalysis of thousands of samples per day. The tasks involved in LC/MS analysis are readily divided into three areas, sample preparation/liquid handling, LC/MS method building/sample analysis, and data processing. Several automation and multitasking strategies were developed and implemented to minimize plating and liquid handling errors, reduce dead times within the analysis cycle, and allow for comprehensive review of data. Delivering multiple samples to multiple injectors allows the autosampler time to complete its wash cycles and aspirate the next set of samples while the previous set is being analyzed. A dual-column chromatography system provides column cycling and peak stacking and allows rapid throughput using conventional LC equipment. Collecting all data for a compound into a single file greatly reduces the number of data files collected, increases the speed of data collection, allows rugged and complete review of all data, and provides facile data management. The described systems have analyzed over 40 000 samples per month for two years and have the capacity for over 2000 samples per instrument per day.

AutoScan: an automated workstation for rapid determination of mass and tandem mass spectrometry conditions for quantitative bioanalytical mass spectrometry.

An automated flow injection analysis (FIA) mass spectrometry system (AutoScan) was developed to allow rapid unattended determination of optimal conditions during mass (ms) and tandem mass spectrometry (ms/ms) on new chemical entities (NCEs) arranged in 96-well plates. The 96-well plate is placed on the deck of a modified Gilson Multiprobe autosampler for injection into a PE Sciex API 2000 triple quadrupole mass spectrometer. A customized software interface is used to create the necessary scan experiments by associating each 96-well plate of NCEs to be scanned with an index file containing data on the identity of each analyte and its expected molecular weight. Analytes are injected four at a time into a custom injection manifold and conventional mass spectra are acquired in both polarities (+/-) using an alternating positive/negative Q1 scan function. The software determines the optimal polarity and definitive precursor ion for all analytes and uses the results to build the injection sequence for product ion scanning. The samples are automatically re-injected under MS/MS conditions, and product ion scans that loop among different collision energies are collected for each analyte. The resulting data are processed automatically and the optimal MS/MS transitions for each analyte are selected. A color-coded graphical interface facilitates data review. Any unusual ion transitions or transposition errors made during plate preparation are noted and corrected. Complete MS and MS/MS conditions are obtained for 96 compounds in about one hour and the resulting data are available for download as sample control injection sequence files.

Arterial hypoxaemia in endurance athletes is greater during running than cycling.

The effect of both training discipline and exercise modality on exercise-induced hypoxaemia (EIH) was examined in seven runners and six cyclists during 5 min high intensity treadmill and cycle exercise. There were no significant interactions between training discipline, exercise modality and arterial P(O(2)) (Pa(O(2))) when subject groups were considered separately but when pooled there were significant differences between exercise modalities. After min 2 of exercise arterial hydrogen ion concentration, minute ventilation, alveolar P(O(2)) (PA(O(2))) and Pa(O(2)) were all lower with treadmill running with the largest differential for the latter occurring at min 5 (treadmill, 80.8+/-1.8; cycle, 90.2+/-2.5, mmHg, N=13, P< or = 0.05). At every min of exercise, the differences in Pa(O(2)) between the ergometers were strongly associated with similar differences in PA(O(2)) and alveolar to arterial P(O(2)) (PA(O(2))-Pa(O(2))). It is concluded that the greater EIH with treadmill running is a consequence of the combined effect of a reduced lactic acidosis-induced hyperventilation and greater ventilation-perfusion inequality with this exercise mode.

A phase II trial of murine monoclonal antibody 17-1A and interferon-gamma: clinical and immunological data.

A group of 15 patients with metastatic colorectal adenocarcinoma received a combination of interferon gamma (0.1 mg/m2, days 1-15) and the murine monoclonal antibody 17-1A (400 mg, days 5, 7, 9 and 12). The treatment was tolerated with minimal toxicity. Of the 14 evaluable patients, 13 developed human antibody to murine 17-1A, with 11 patients demonstrating antibody to the variable region of 17-1A (anti-idiotype). Antibody to the variable region was inhibited by 17-1A but not by mouse immunoglobulin. Sera from patients with substantial anti-idiotype reactivity were capable of inhibiting the binding of murine 17-1A to antigen expressing LS174-T cells thus indicating the presence of antibody directed against the 17-1A combining site (mirror-image anti-idiotype). The median survival of the whole group was 56 weeks and there was no correlation between clinical response/survival and the development of anti-idiotype antibody.

Quantitative assay of a human monoclonal IgM antibody (HA-1A) in human serum.

A rapid, specific and sensitive radiometric assay was developed capable of quantitating serum levels of HA-1A, a human IgM monoclonal antibody to endotoxin. 'Private' anti-idiotypic murine monoclonal antibodies were produced and utilized in the assay to avoid cross-reactivity with normal human IgG, IgM, IgA, IgE or IgD. The presence of E. coli or gram-negative lipopolysaccharide in the sera did not affect the ability of the assay to detect HA-1A. The sensitivity of the assay was calculated to be 25 ng/ml with an interassay coefficient of variation of less than 10%. In one patient given 100 mg of HA-1A, peak serum concentration was 101.5% of the predicted value with a mean plasma half life of 24.5 h. This assay will be useful in establishing the pharmacokinetics of HA-1A and in monitoring serum levels during phase II and phase III clinical trials.

Development and characterization of a monoclonal antibody to human embryonal carcinoma.

A monoclonal anti-testicular carcinoma antibody was obtained via the somatic cell fusion technique by immunization of BALB/c mice with freshly prepared single cell suspension from a patient with testicular embryonal carcinoma with choriocarcinoma components. The hybridoma supernates were screened against the testicular carcinoma cells used in the immunization as well as normal mononuclear white blood cells isolated from the same patient. An antibody (5F9) was selected which bound to fresh tumor cells from two patients with embryonal testicular carcinoma and failed to bind to fresh tumor cells from 24 patients (2 seminoma, 2 melanoma, 3 neck, 2 esophageal, 1 ovarian, 3 colon, 1 prostate, 2 breast, 1 liposarcoma, 3 endometrial, 1 kidney, 1 adrenal, 1 larynx and 1 bladder tumors) or cell suspensions prepared from normal liver, lung, spleen, ovary, testes, kidney, red blood cells or white blood cells. The antibody was tested for its binding to several well established cancer cell lines, and was found to bind to the BeWo human choriocarcinoma and two human embryonal carcinoma cell lines. The antibody did not react with 22 other cell lines or with hCG. The antibody was labeled with 131I and injected into nude mice bearing BeWo tumors and evaluated for tumor localization by performing whole body scans with a gamma camera 5 days later. Six mice injected with the antibody showed positive tumor localization without the need for background subtraction while six mice injected with MOPC-21, a murine myeloma immunoglobulin, demonstrated much less tumor localization. Tissue distribution studies performed after scanning showed specific tumor localization (8:1 tumor: muscle) for the monoclonal antibody and no specific localization for MOPC-21. This antibody thus has selective reactivity with the surface of tumor cells from embryonal carcinoma (testicle) and choriocarcinoma both in vitro and in vivo.

Sonographic detection of fibromatosis: a rare tumor in the retroperitoneum.

Abdominal sonography for the detection and evaluation of mass lesions is a common investigative modality. The authors wish to report a case of an unusual retroperitoneal tumor that was detected by ultrasound. The lesion, fibromatosis, was symptomatic but went undiagnosed for over a year, despite two evaluations which included multiple standard roentgenographic studies. Fibromatosis is discussed.

Effect of hypophysectomy on persistence of methylated purines in rat liver deoxyribonucleic acid after administration of dimethylnitrosamine.

The formation of methylated purines in DNA following dimethylnitrosamine administration was studied in control and hypophysectomized rats. When given the same dose of this carcinogen (in mg/kg body weight) the formation of the major product 7-methylguanine and of the minor products 1-, 3- and 7-methyladenine and 3-methylguanine was slightly greater in the livers of hypophysectomized rats than in controls. The rate of loss of these products from the DNA was not affected by hypophysectomy. O6-Methylguanine levels were significantly greater in the hepatic DNA of hypophysectomized rats compared to controls after doses of dimethylnitrosamine ranging from 1 to 20 mg/kg. This difference was due to a slower rate of loss of this purine from the DNA in the hypothysectomized rats. Growth hormone treatment increased the rate of removal of O6-methylguanine in the hypophysectomized rats but did not restore the activity to that found in controls. The possible significance of these results in the induction of tumors by dimethylnitrosamine is discussed.