Micro-punches versus micro-slices for serial sampling of human dentine: Striking a balance between improved temporal resolution and measuring additional isotope systems.

RATIONALE: The last decade has seen a dramatic increase in the application of serial sampling of human dentine in archaeology. Rapid development in the field has provided many improvements in the methodology, in terms of both time resolution as well as the ability to integrate more isotope systems in the analysis. This study provides a comparison of two common sampling approaches, allowing researchers to select the most suitable approach for addressing specific research questions. METHODS: Two common approaches for sequential sampling of human dentine (micro-punches and micro-slices) are compared in terms of viability and efficacy. Using archaeological deciduous second molars and permanent first molars, this study demonstrates how the two approaches capture aspects of the weaning process in different ways. In addition, different aspects related to the extraction protocols, such as the thickness of the central slide and the solubilisation step, are also evaluated. RESULTS: While both approaches show similar intra-tooth isotopic patterns, the micro-punches approach is preferable for research that requires a very fine temporal resolution, while the micro-slices approach is best for research where δ34 S values are needed, or when the samples are poorly preserved. In addition, the solubilisation step has a large effect on collagen yield, and, to a lesser extent, on isotopic compositions. Therefore, it is important to ensure that only samples that have undergone the same pre-treatment protocol are directly compared. CONCLUSIONS: We present the pros and cons of the two micro-sampling approaches and offer possible mitigation strategies to address some of the most important issues related to each approach.

Design, synthesis and structure-activity relationships of substituted oxazole-benzamide antibacterial inhibitors of FtsZ.

The design, synthesis and structure-activity relationships of a series of oxazole-benzamide inhibitors of the essential bacterial cell division protein FtsZ are described. Compounds had potent anti-staphylococcal activity and inhibited the cytokinesis of the clinically-significant bacterial pathogen Staphylococcus aureus. Selected analogues possessing a 5-halo oxazole also inhibited a strain of S. aureus harbouring the glycine-to-alanine amino acid substitution at residue 196 of FtsZ which conferred resistance to previously reported inhibitors in the series. Substitutions to the pseudo-benzylic carbon of the scaffold improved the pharmacokinetic properties by increasing metabolic stability and provided a mechanism for creating pro-drugs. Combining multiple substitutions based on the findings reported in this study has provided small-molecule inhibitors of FtsZ with enhanced in vitro and in vivo antibacterial efficacy.

Environmental pH affects photoautotrophic growth of Synechocystis sp. PCC 6803 strains carrying mutations in the lumenal proteins of PSII.

Synechocystis sp. strain PCC 6803 grows photoautotrophically across a broad pH range, but wild-type cultures reach a higher density at elevated pH; however, photoheterotrophic growth is similar at high and neutral pH. A number of PSII mutants each lacking at least one lumenal extrinsic protein, and carrying a second PSII lumenal mutation, are able to grow photoautotrophically in BG-11 medium at pH 10.0, but not pH 7.5. We investigated the basis of this pH effect and observed no pH-specific change in variable fluorescence yield from PSII centers of the wild type or the pH-dependent ΔPsbO:ΔPsbU and ΔPsbV:ΔCyanoQ strains; however, 77 K fluorescence emission spectra indicated increased coupling of the phycobilisome (PBS) antenna at pH 10.0 in all mutants. DNA microarray data showed a cell-wide response to transfer from pH 10.0 to pH 7.5, including decreased mRNA levels of a number of oxidative stress-responsive transcripts. We hypothesize that this transcriptional response led to increased tolerance against reactive oxygen species and in particular singlet oxygen. This response enabled photoautotrophic growth of the PSII mutants at pH 10.0. This hypothesis was supported by increased resistance of all strains to rose bengal at pH 10.0 compared with pH 7.5.

An improved small-molecule inhibitor of FtsZ with superior in vitro potency, drug-like properties, and in vivo efficacy.

The bacterial cell division protein FtsZ is an attractive target for small-molecule antibacterial drug discovery. Derivatives of 3-methoxybenzamide, including compound PC190723, have been reported to be potent and selective antistaphylococcal agents which exert their effects through the disruption of intracellular FtsZ function. Here, we report the further optimization of 3-methoxybenzamide derivatives towards a drug candidate. The in vitro and in vivo characterization of a more advanced lead compound, designated compound 1, is described. Compound 1 was potently antibacterial, with an average MIC of 0.12 µg/ml against all staphylococcal species, including methicillin- and multidrug-resistant Staphylococcus aureus and Staphylococcus epidermidis. Compound 1 inhibited an S. aureus strain carrying the G196A mutation in FtsZ, which confers resistance to PC190723. Like PC190723, compound 1 acted on whole bacterial cells by blocking cytokinesis. No interactions between compound 1 and a diverse panel of antibiotics were measured in checkerboard experiments. Compound 1 displayed suitable in vitro pharmaceutical properties and a favorable in vivo pharmacokinetic profile following intravenous and oral administration, with a calculated bioavailability of 82.0% in mice. Compound 1 demonstrated efficacy in a murine model of systemic S. aureus infection and caused a significant decrease in the bacterial load in the thigh infection model. A greater reduction in the number of S. aureus cells recovered from infected thighs, equivalent to 3.68 log units, than in those recovered from controls was achieved using a succinate prodrug of compound 1, which was designated compound 2. In summary, optimized derivatives of 3-methoxybenzamide may yield a first-in-class FtsZ inhibitor for the treatment of antibiotic-resistant staphylococcal infections.

Early cystic fibrosis lung disease detected by bronchoalveolar lavage and lung clearance index.

RATIONALE: Unrecognized airway infection and inflammation in young children with cystic fibrosis (CF) may lead to irreversible lung disease; therefore early detection and treatment is highly desirable. OBJECTIVES: To determine whether the lung clearance index (LCI) is a sensitive and repeatable noninvasive measure of airway infection and inflammation in newborn-screened children with CF. METHODS: Forty-seven well children with CF (mean age, 1.55 yr) and 25 healthy children (mean age, 1.26 yr) underwent multiple-breath washout testing. LCI within and between-test variability was assessed. Children with CF also had surveillance bronchoalveolar lavage performed. MEASUREMENTS AND MAIN RESULTS: The mean (SD) LCI in healthy children was 6.45 (0.49). The LCI was higher in children with CF (7.21 [0.81]; P < 0.001). The upper limit of normal for the LCI was 7.41. Fifteen (32%) children with CF had an elevated LCI. LCI measurements were repeatable and reproducible. Airway infection was present in 17 (36%) children with CF, including 7 (15%) with Pseudomonas aeruginosa. Polymicrobial growth was associated with worse inflammation. The LCI was higher in children with Pseudomonas (7.92 [1.16]) than in children without Pseudomonas (7.02 [0.56]) (P = 0.038). The LCI correlated with bronchoalveolar lavage IL-8 (R(2) = 0.20, P = 0.004) and neutrophil count (R(2) = 0.21, P = 0.001). An LCI below the upper limit of normality had a high negative predictive value (93%) in excluding Pseudomonas. CONCLUSIONS: The LCI is elevated early in CF, especially in the presence of Pseudomonas and airway inflammation. The LCI is a feasible, repeatable, and sensitive noninvasive marker of lung disease in young children with CF.

Teaching Joint Attention: Assessing Generalization and Maintenance of Effects using Multiple Exemplar Training.

The purpose of the present study was to evaluate the effects of multiple exemplar training and social reinforcement on the maintenance and generalization of joint attention initiations across toy classes. Three children with autism spectrum disorder (ASD) participated. After analyzing samples of joint attention initiations from neurotypical peers, a composite score was developed and used to evaluate joint attention initiations of the children with ASD. Gaze shifting and commenting were taught using social reinforcement in a multiple exemplar format. Training sessions were followed by probes of untrained stimuli both within and across toy classes. Results showed that the participants acquired both gaze shifting and commenting using social consequences as reinforcers. Following training, participants initiated joint attention with a frequency within the range of their typically developing peers. Multiple exemplar training was also effective in facilitating acquisition within classes and joint attention maintenance during follow-up probes. Additionally, all participants generalized the acquired skills to a class of untrained stimuli.

Stereotypy in young children with autism and typically developing children.

Although stereotypy is one of the key diagnostic features of autism, few studies have compared stereotypic behavior in children with autism and typically developing children. The present study employed direct observational measurement methods to assess levels of stereotypic behavior in 2-, 3- and 4-year-old children with autism or pervasive developmental disorder - not otherwise specified (PDD-NOS) and age-matched typically developing peers. Thirty children with autism or PDD-NOS and 30 typically developing children participated. Each child's performance of several early learning and play skills was assessed using a direct observational assessment protocol developed for children with autism who were entering early intensive behavioral treatment. Duration of episodes of vocal and motor stereotypy was recorded from a videotaped 10 min portion of that assessment session. Results indicated that the 2-year-old children with autism or PDD-NOS had somewhat higher levels of stereotypic behavior than the typically developing 2-year-olds, while the 3- and 4-year-old children with autism or PDD-NOS displayed substantially higher levels stereotypic behavior than their same-age peers.

Assessing and treating vocal stereotypy in children with autism.

Previous research implies that stereotypic behavior tends to be maintained by the sensory consequences produced by engaging in the response. Few investigations, however, have focused on vocal stereotypy. The current study examined the noncommunicative vocalizations of 4 children with an autism spectrum disorder. First, functional analyses were conducted in an attempt to identify the function of each child's behavior. For each of the participants, it was found that vocal stereotypy was likely not maintained by the social consequences. Following assessment, response interruption and redirection (RIRD) was implemented in an ABAB design to determine whether vocal stereotypy could be successfully redirected. RIRD involved a teacher issuing a series of vocal demands the child readily complied with during regular academic programming. Vocal demands were presented contingent on the occurrence of vocal stereotypy and were continuously presented until the child complied with three consecutively issued demands without emitting vocal stereotypy. For each child, RIRD produced levels of vocal stereotypy substantially lower than those observed in baseline. For 3 of the children, an increase in appropriate communication was also observed. The children's teachers were trained to implement RIRD. Brief follow-up probes and anecdotal information implied that the treatment had a positive impact in the natural environment.

Behavioral assessment of joint attention: a methodological report.

This paper describes a highly structured assessment protocol with objective behavioral measures for joint attention responding and initiation. The assessment was given to 26 children diagnosed with autism spectrum disorders and 21 typically developing children, aged two to four years. Interobserver agreement was high for all behavioral measures. Children with autism had relatively minor deficits in joint attention responding and more severe deficits in joint attention initiation, relative to typically developing children. These results replicate those reported in previous research. The protocol can be used reliably to assess behavior indicative of joint attention responding and initiation in typically developing children and children with autism.

Taking Full Responsibility: the Ethics of Supervision in Behavior Analytic Practice.

The supervision of professionals in the field of behavior analysis is multifaceted. The BACB® Professional and Ethical Compliance Code for Behavior Analysis provides guidance for effective supervisory practices, as supervision impacts both the supervisee and the consumers. The purpose of this article is (1) to discuss rationales and consequences relative to supervision issues, (2) to provide directions for professional development in each of the seven identified supervisory areas within the code, and (3) to set the occasion for critical discourse relative to supervision. Case examples are used to illustrate each of the seven supervisory subcomponents of the "Behavior Analysts as Supervisors" section of the Code. A rationale is provided for each component, as well as a discussion of possible undesirable consequences resulting from not following the rule. While the code provides clear expectations of the desired behavior, this article explores more of the subtle nuances inherent in each section of the supervision code, with the goal of achieving a better understanding of the Code and enhancing supervisory skills.

Lake Ontario salmon (Salmo salar) were not migratory: A long-standing historical debate solved through stable isotope analysis.

Lake Ontario once supported a large complex of Atlantic Salmon (Salmo salar) populations that became extinct prior to scientific study. Since the 1860s, research efforts to conserve and reintroduce a sustainable population of Atlantic Salmon have focused on determining whether Lake Ontario's original salmon populations had migrated to the Atlantic Ocean as part of their lifecycle (anadromy), stayed in the lake year-round (potamodromy), or both. We used stable carbon, nitrogen, and sulfur isotope analyses of archaeological bones and historical museum-archived salmon scales to show that the original salmon populations from Lake Ontario completed their entire lifecycle without migrating to the Atlantic Ocean. With a time depth of more than 500 years, our findings provide a unique baseline with significant potential for informing modern restocking and conservation efforts.

Collagen-carbon nanotube composite materials as scaffolds in tissue engineering.

Carbon nanotubes (CNT) are attractive for use in fiber-reinforced composite materials due to their very high aspect ratio, combined with outstanding mechanical and electrical properties. Composite materials comprising a collagen matrix with embedded CNT were prepared by mixing solubilized Type I collagen with solutions of carboxylated single-walled carbon nanotubes (SWNT) at concentrations of 0, 0.2, 0.4, 0.8, and 2.0 weight percent. Living smooth muscle cells were incorporated at the time of collagen gelation to produce cell-seeded collagen-CNT composite matrices. Constructs containing 2.0 wt % CNT exhibited delayed gel compaction, relative to lower concentrations that compacted at the same rate as pure collagen controls. Cell viability in all constructs was consistently above 85% at both Day 3 and Day 7, whereas cell number in CNT-containing constructs was lower than in control constructs at Day 3, though statistically unchanged by Day 7. Scanning electron microscopy showed physical interactions between CNT and collagen matrix. Raman spectroscopy confirmed the presence of CNT at the expected diameter (0.85-1.30 nm), but did not indicate strong molecular interactions between the collagen and CNT components. Such collagen-CNT composite matrices may have utility as scaffolds in tissue engineering, or as components of biosensors or other medical devices.

Comparison of direct observational methods for measuring stereotypic behavior in children with autism spectrum disorders.

We compared partial-interval recording (PIR) and momentary time sampling (MTS) estimates against continuous measures of the actual durations of stereotypic behavior in young children with autism or pervasive developmental disorder-not otherwise specified. Twenty-two videotaped samples of stereotypy were scored using a low-tech duration recording method, and relative durations (i.e., proportions of observation periods consumed by stereotypy) were calculated. Then 10, 20, and 30s MTS and 10s PIR estimates of relative durations were derived from the raw duration data. Across all samples, PIR was found to grossly overestimate the relative duration of stereotypy. Momentary time sampling both over- and under-estimated the relative duration of stereotypy, but with much smaller errors than PIR (Experiment 1). These results were replicated across 27 samples of low, moderate and high levels of stereotypy (Experiment 2).

Toward a behavioral analysis of joint attention.

Joint attention (JA) initiation is defined in cognitive-developmental psychology as a child's actions that verify or produce simultaneous attending by that child and an adult to some object or event in the environment so that both may experience the object or event together. This paper presents a contingency analysis of gaze shift in JA initiation. The analysis describes reinforcer-establishing and evocative effects of antecedent objects or events, discriminative and conditioned reinforcing functions of stimuli generated by adult behavior, and socially mediated reinforcers that may maintain JA behavior. A functional analysis of JA may describe multiple operant classes. The paper concludes with a discussion of JA deficits in children with autism spectrum disorders and suggestions for research and treatment.

Assessing progress and outcome of early intensive behavioral intervention for toddlers with autism.

Intensive behavioral intervention for young children diagnosed with autism can produce large gains in social, cognitive, and language development. Although several studies have identified behaviors that are possible indicators of best outcome, changes in performance are typically measured using norm-referenced standardized scores referencing overall functioning level rather than via repeated observational measures of autism-specific deficits (i.e., social behavior). In the current study, 83 children with autism (CWA), aged 1, 2 and 3 years, and 58 same-aged typically developing children (TDC) were directly observed in the areas of cognitive skills, joint attention (JA), play, and stereotypic behavior using a measure called the Early Skills Assessment Tool (ESAT; MacDonald et al., 2006). CWA were assessed at entry into an EIBI program and again after 1 year of treatment. Changes in performance were compared pre- and post-treatment as well as to the normative data by age. Results indicate significant gains on the ESAT across all age groups with the greatest gains seen in the children who entered treatment prior to their second birthday. Increases were seen on direct measures of JA, play, imitation and language while decreases were seen in stereotypy regardless of level of performance at entry into EIBI. The ESAT, a direct measurement tool, served as a sensitive tool to measure changes in autism symptomatology following EIBI treatment.

Social validation of the New England Center For Children--Core Skills Assessment.

We investigated the social validity of the NECC Core Skills Assessment (NECC-CSA) with parents and professionals as participants. The NECC-CSA is a measurement tool consisting of direct and indirect measures of skills important to all individuals with autism, across the lifespan. Participants (N = 245) were provided with a list of 66 skills, 47 of which were Core Skills from the NECC-CSA, and were asked to indicate which items they considered to be foundational. Participants endorsed items from the NECC-CSA as foundational skills, more than they endorsed the other items. Differences between parents and professionals are described and detailed with respect to individual assessment items. The NECC-CSA consists of socially validated skills that can be taken as a starting point for programs of instruction for individuals with ASDs.

Using video modeling with substitutable loops to teach varied play to children with autism.

Children with autism often engage in repetitive play with little variation in the actions performed or items used. This study examined the use of video modeling with scripted substitutable loops on children's pretend play with trained and untrained characters. Three young children with autism were shown a video model of scripted toy play that included a substitutable loop that allowed various characters to perform the same actions and vocalizations. Three characters were modeled with the substitutable loop during training sessions, and 3 additional characters were present in the video but never modeled. Following video modeling, all the participants incorporated untrained characters into their play, but the extent to which they did so varied.

Creating an antibacterial with in vivo efficacy: synthesis and characterization of potent inhibitors of the bacterial cell division protein FtsZ with improved pharmaceutical properties.

3-Methoxybenzamide (1) is a weak inhibitor of the essential bacterial cell division protein FtsZ. Alkyl derivatives of 1 are potent antistaphylococcal compounds with suboptimal drug-like properties. Exploration of the structure-activity relationships of analogues of these inhibitors led to the identification of potent antistaphylococcal compounds with improved pharmaceutical properties.

Using video modeling to teach reciprocal pretend play to children with autism.

The purpose of the present study was to use video modeling to teach children with autism to engage in reciprocal pretend play with typically developing peers. Scripted play scenarios involving various verbalizations and play actions with adults as models were videotaped. Two children with autism were each paired with a typically developing child, and a multiple-probe design across three play sets was used to evaluate the effects of the video modeling procedure. Results indicated that both children with autism and the typically developing peers acquired the sequences of scripted verbalizations and play actions quickly and maintained this performance during follow-up probes. In addition, probes indicated an increase in the mean number of unscripted verbalizations as well as reciprocal verbal interactions and cooperative play. These findings are discussed as they relate to the development of reciprocal pretend-play repertoires in young children with autism.

Teaching Discrimination of Adult Gaze Direction to Children with Autism.

Three young children diagnosed with autism did not reliably locate objects in the environment on the basis of an adult's gaze shifts. A training program designed to teach gaze following used the activation of remote controlled mechanical toys as both prompts and consequences. Over several training sessions, toy activation was progressively delayed following the adult's gaze-shift cues. All of the children eventually came to anticipate the toy activation and locate the target object on the basis of the adult's gaze-shift cue alone. Discrimination of another person's gaze direction is discussed in relation to joint attention deficits in children with autism.