Occurrence and characterization of Escherichia coli ST410 co-harbouring blaNDM-5, blaCMY-42 and blaTEM-190 in a dog from the UK.

BACKGROUND/OBJECTIVES: Carbapenemase-producing Enterobacteriaceae (CPE) are a public health threat, and have been found in humans, animals and the environment. Carbapenems are not authorized for use in EU or UK companion animals, and the prevalence of carbapenem-resistant Gram-negative bacilli (CRGNB) in this population is unknown. METHODS: We investigated CRGNB isolated from animal specimens received by one diagnostic laboratory from 34 UK veterinary practices (September 2015-December 2016). Any Gram-negative isolates from clinical specimens showing reduced susceptibility to fluoroquinolones and/or aminoglycosides and/or cephalosporins were investigated phenotypically and genotypically for carbapenemases. A complete genome assembly (Illumina/Nanopore) was generated for the single isolate identified to investigate the genetic context for carbapenem resistance. RESULTS: One ST410 Escherichia coli isolate [(CARB35); 1/191, 0.5%], cultured from a wound in a springer spaniel, harboured a known carbapenem resistance gene (blaNDM-5). The gene was located in the chromosome on an integrated 100 kb IncF plasmid, also harbouring other drug resistance genes (mrx, sul1, ant1 and dfrA). The isolate also contained blaCMY-42 and blaTEM-190 on two separate plasmids (IncI1 and IncFII, respectively) that showed homology with other publicly available plasmid sequences from Italy and Myanmar. CONCLUSIONS: Even though the use of carbapenems in companion animals is restricted, the concurrent presence of blaCMY-42 and other antimicrobial resistance genes could lead to co-selection of carbapenemase genes in this population. Further studies investigating the selection and flow of plasmids carrying important resistance genes amongst humans and companion animals are needed.

Preclinical absorption, distribution, metabolism and excretion (ADME) characterization of ICAM1988, an LFA-1/ICAM antagonist, and its prodrug.

Intercellular adhesion molecule (ICAM)-1988 is a small molecule lymphocyte function-associated antigen-1 (LFA-1) antagonist being considered for its anti-inflammatory properties. Following intravenous administration of ICAM1988, clearances in mice, rats, dogs, and monkeys were 17.8, 3.31, 15.4, and 6.85 ml min(-1) kg(-1), respectively. In mass balance studies using [(14)C]-ICAM1988 in rats dosed intravenously, unchanged ICAM1988 contributed to 25.1% of the dose. In rats, the systemic bioavailability of ICAM1988 was improved to 0.28 when the drug was administered orally as its isobutyl ester, ICAM2660. In rats, this was consistent with the complete in vitro conversion of ICAM2660 to ICAM1988 in plasma, and liver and intestinal S9. In dogs and monkeys, ICAM2660 did not improve the bioavailability of ICAM1988. This is consistent with limited in vitro conversion of ICAM2660 to ICAM1988 in plasma and liver S9. In human in vitro studies, ICAM2660 conversion to ICAM1988 in liver was similar to rats while no conversion in plasma and intestinal S9 fractions were observed. Based on the in vitro metabolism similarities of human and rat, it would be anticipated that in human oral administration of ICAM2660 would improve the systemic exposure of ICAM1988.

Generation of an LFA-1 antagonist by the transfer of the ICAM-1 immunoregulatory epitope to a small molecule.

The protein-protein interaction between leukocyte functional antigen-1 (LFA-1) and intercellular adhesion molecule-1 (ICAM-1) is critical to lymphocyte and immune system function. Here, we report on the transfer of the contiguous, nonlinear epitope of ICAM-1, responsible for its association with LFA-1, to a small-molecule framework. These LFA-1 antagonists bound LFA-1, blocked binding of ICAM-1, and inhibited a mixed lymphocyte reaction (MLR) with potency significantly greater than that of cyclosporine A. Furthermore, in comparison to an antibody to LFA-1, they exhibited significant anti-inflammatory effects in vivo. These results demonstrate the utility of small-molecule mimics of nonlinear protein epitopes and the protein epitopes themselves as leads in the identification of novel pharmaceutical agents.

Presentation rate in comprehension of natural and synthesized speech.

This study examined the effect of four presentation rates (approximately 130, 150, 170, and 190 words per minute) on the comprehension of natural and synthesized speech by having 96 subjects [1 man, 95 women ranging in age from 18 to 40 years (M=21.0)] perform a sentence-verification task. Analysis showed that, while their response latencies were significantly faster to natural than to synthesized speech, presentation rate did not have a significant effect on response latencies when sentences were presented at rates within the average speaking range (approximately 130 to 190 words per minute). Implications these findings may have for the use of synthesized speech in human factors applications are discussed.

A novel family of hairpin peptides that inhibit IgE activity by binding to the high-affinity IgE receptor.

A family of structured peptides that bind to FcepsilonRIalpha, the alpha-chain of the high-affinity receptor for IgE, has been identified. Binding selections using FcepsilonRIalpha and polyvalent peptide-phage libraries yielded a dominant 18-residue peptide-phage clone, as well as related sequences that did not resemble fragments of IgE. Synthetic peptides based on these sequences inhibited IgE binding to its receptor, and were found by NMR analysis to adopt a stable beta-hairpin structure in solution. Optimized peptides with micromolar receptor affinity exhibited high stability in biological fluids and inhibited cellular histamine release in an in vitro bioassay of IgE activity. The structure-activity relationships of these peptides, which are less than 1% of the size of IgE, suggest an overlap between their binding site and that of IgE on FcepsilonRI. Thus, the peptides demonstrate that blocking a small epitope on this receptor chain is sufficient to block IgE activity. Such structured peptides represent a possible starting point for the design of novel antagonists, and offer the potential for testing in vivo a new approach for treating allergic disease.

Behavioral health care: the keys to successful planning and management.

Behavioral health care is an important component of health system development. Unfortunately, many systems believe it is unmanageable in relation to both cost and effectiveness. The keys to success include thorough planning and modernized oversight procedures.

Growth inhibition of subcutaneously transplanted hepatomas without cachexia by alteration of the dietary arginine-methionine balance.

Previous studies have shown that alteration of the dietary arginine-methionine balance by use of synthetic L-amino acids inhibits tumor growth of a subcutaneously transplanted Morris hepatoma at the expense of maintaining body weight. However, L-methionine is susceptible to degradation and, therefore, may contribute to a deficiency state. The present studies were performed to determine whether growth of subcutaneous hepatoma transplants is inhibited, and body growth maintained, when rats are fed diets containing L-methionine in replacement of N-acetyl-L-methionine (NALM) for 28 days. Tumor-free and tumor-bearing rats fed a control diet, with amino acids replacing protein, had gains in body weight: 31.3 +/- 1.0 and 19.1 +/- 0.5 g (12% and 7%), respectively. Rats fed six experimental diets, with varying L-arginine-NALM balances, had body weight gains ranging from 18.4 +/- 0.3 to 26.7 +/- 0.9 g (7-10%). Tumor weight of control rats was 10.65 +/- 0.24% of body weight. Diets supplemented with L-arginine in combination with normal and deficient NALM decreased tumor weights by 35% and 38%, respectively, It is concluded that dietary replacement of L-methionine with NALM and supplementation with L-arginine inhibits growth of a subcutaneously transplanted Morris hepatoma in the absence of cachexia.

Synthetic speech comprehension: a comparison of children with normal and impaired language skills.

This study compared the ability of children with normal language (NL) and children with specific language impairment (SLI) to comprehend natural speech and DECtalk synthetic speech by using a sentence verification task. The effect of listening practice on subjects' ability to comprehend both types of speech also was investigated. Subjects were matched for age and sex. Mean nonverbal intelligence scores of the groups did not differ significantly. Results showed that DECtalk was significantly more difficult for all subjects to comprehend than was natural speech and false sentences were significantly more difficult to comprehend than were true sentences. Response latencies shortened significantly from time 1 to time 2 for all subjects. Subjects with SLI had significantly more difficulty comprehending both natural and synthetic speech than did subjects with NL. Implications these results might have for theories of the underlying cause of specific language impairment are discussed.

Effect of visual cuing on synthetic speech intelligibility: a comparison of native and nonnative speakers of English.

This study compared the effect of visual cuing on the intelligibility of DECtalk for native and nonnative speakers of English in both ideal listening conditions and in the presence of background noise at a signal to noise (S/N) ratio of + 10dB. Visual cuing improved DECtalk's intelligibility for nonnative speakers more than for native speakers, especially in the background noise condition. Implications of these findings and the need for further research are discussed.

Growth inhibition of subcutaneously transplanted hepatomas by alterations of the dietary arginine-methionine balance.

We hypothesized that alteration of the dietary arginine-methionine balance might inhibit tumor growth and suggest nutritional strategies for cancer therapy. The Morris hepatoma 3924A was subcutaneously transplanted in ACI rats. Control diets containing normal levels of arginine, methionine, and other amino acids in replacement of protein (24%), carbohydrates (59%), fat (10%), and fiber, vitamins, and minerals (7%) were fed for 28 days. Six experimental diets were adjusted to maintain amino acids at 23-25% and carbohydrates at 58-60%; these diets were 1%-2% deficient in arginine or supplemented with 1-2% arginine (expressed as percent amino acid content of diet) in combination with normal, deficient, and supplementary levels of methionine. Daily food intake was unaffected by the experimental diets. The control groups gained 26.4 +/- 2.8 g body weight, and small body weight decrements ranged from 3.5% to 8.4% in the groups fed the experimental diets. Tumor weight of controls was 8.5 +/- 1.5% of body weight. The experimental diets that produced significant tumor growth inhibition (TGI) were 1) the arginine-methionine-deficient diet, 2) the arginine-excess-methionine-deficient diet, 3) the arginine-deficient diet, and 4) the excess-arginine diet. Diets containing excess methionine failed to produce TGI. TGI resulted in tumor weights 41-46% of control values. TGI was associated with significantly lower blood urea nitrogen, plasma protein, and tumor spermidine-to-spermine ratio than in tumor-bearing controls. It is concluded that dietary alteration of a single amino acid, arginine, might be a potentially useful nutritional strategy for controlling tumor growth.

Language familiarity in magnitude-estimation scaling of loudness by young adults.

The purpose was to examine the effect of language familiarity on magnitude-estimation scaling of loudness by young adults. Two groups of subjects participated in this study. Group 1 of 20 subjects (M age = 23.95 yr.) were familiar with English and not familiar with Hindi. Group 2 of 20 subjects (M age = 24.30 yr.) were familiar with English as well as Hindi. Two separate magnitude-estimation scaling tasks were performed. On the first scaling task, an English sentence was used as the stimulus, and on the second scaling task, a Hindi sentence was used as the stimulus. Statistical analysis showed that there was no significant difference between the two groups in loudness judgments of the English and Hindi sentences. Subjects scaled the loudness of an unfamiliar language in the same manner as they scaled the loudness of a familiar language. The findings suggest that magnitude-estimation scaling is an effective measure of loudness whether the language being listened to is familiar to the listener.

An examination of alveolar stop retraction during pacifier use.

Employing a single subject A-B-A experimental design, a four year old female with normal articulation and a history of pacifier use was observed to demonstrate alveolar stop retraction more than fifty percent of the time while using her orthodontic device. Contextual factors appeared to have played a role in this variability. Upon confirming this phenomenon in the four year old subject, a follow-up investigation was undertaken with seven children between the ages of three and five years. All subjects had age appropriate articulation skills with no evidence of alveolar stop retraction without the pacifier in place and only rarely with the pacifier in place (.85%). Implications of these findings and the need for further research are discussed.

Benzodiazepine peptidomimetic inhibitors of farnesyltransferase.

A structural survey of protein Zn2+ binding geometries was instigated based upon the functional requirement of Ras farnesyltransferase for Zn2+. The Cys-X-X-Cys motif found in Zn(2+)-binding proteins such as aspartate transcarbamylase was used as a template to devise a bidentate-coordination model for Cys-A1-A2-X peptide inhibitors. Accordingly, replacement of the central dipeptide with the hydrophobic scaffold 3-amino-1-carboxymethyl-2,3-dihydro-5- phenyl-1H-1,4-benzodiazepin-2-one (BZA) yielded a peptidomimetic inhibitor, Cys(BZA)Met, of moderate potency (IC50 = 400 nM). N-Methylation of the cysteine amide improved potency almost 100-fold (IC50 = 0.3-1 nM). The increased affinity presumably correlates with a preferred conformation of the inhibitor which maximizes a hydrophobic interaction between the scaffold and the enzyme, and the proper presentation of cysteine and methionine to allow bidentate coordination at Zn2+. These non-peptide inhibitors have been shown to block farnesylation of the Ras protein in intact cells and provide lead compounds for the development of new cancer therapeutic agents.

Localization of nerve growth factor receptor mRNA in contused rat spinal cord by in situ hybridization.

Northern blot analysis using a probe for the low-affinity nerve growth factor receptor (NGFR) revealed that a mild contusive injury induces the expression of NGFR mRNA in rat spinal cord with a maximal expression at 7 days post-injury. We have now localized this induction using in situ hybridization and found the highest concentration of NGFR mRNA at the lesion epicenter. The location and pattern of autoradiographic grains were compared with that of various cell types at the injury site as determined by immunocytochemical studies. The results suggest that cells associated with blood vessels at the epicenter are induced to express NGFR mRNA at 7 days post-injury.

Monocyte esterase deficiency in malignant neoplasia.

A survey of the incidence of monocyte esterase deficiency in 4000 inpatients (including 808 with malignant neoplastic disease) and 474 normal controls was performed using an automated esterase method. A highly significant excess of patients with malignant disease and the deficiency was evident when compared with normal controls or all other patients. Within the group of patients with malignant disease the demonstrable excess occurred in B chronic lymphocytic leukaemia, non-Hodgkin's and Hodgkin's lymphoma, and carcinoma of the gastrointestinal tract. There was also a significant excess of patients with the deficiency attending the renal unit, both among patients who had had renal transplants and those who had not. A familial incidence of monocyte esterase deficiency was found in 19 (35%) of first degree relatives of those patients in whom family studies were done. It is suggested that the reason for the increased prevalence of the anomaly in these disorders might be that the diminution of esterase activity has a role in their development.