Can you see what I see? A comparison of client and observer perspectives of the alliance and group cohesion in CBT.

Both therapeutic alliance and group cohesion have been identified as "demonstrably effective" relationship elements in therapy. However, the overwhelming majority of process-outcome research has relied on clients as raters of the therapeutic relationship. A lack of convergence between client, therapist, and observer perspectives has raised questions regarding how best to measure relationships in therapy. Interest in observational measures has grown, as they may offer more objective and reliable measurements of process. This study compared the predictive validity of client and observer ratings of the alliance (Agreement and Bond) and group cohesion in the context of group cognitive behaviour therapy (CBT) for anxiety disorders. Results showed that client and observer ratings of process were not significantly correlated, and regarding the alliance, only client-rated Agreement predicted client-rated treatment gains. In contrast, both client and observer-ratings of group cohesion were found to uniquely contribute to treatment outcomes. If replicated, the findings from the present study suggest that (1) while client ratings of alliance consistently predict client-rated outcomes, the predictive validity of observer measures has yet to be established, and (2) both clients and observers provide meaningful and distinct information about group cohesion in therapy.

Which, who and when? Therapeutic processes in group cognitive behaviour therapy for anxiety disorders.

The evidence for the effect of therapeutic alliance in group cognitive behaviour therapy (CBT) for anxiety disorders is unclear. Identifying whether the alliance-outcome relationship depends on (1) which components are assessed, (2) who is measuring the alliance and (3) when the alliance is measured will help to clarify the role of the client-therapist relationship in therapy. The present study explored the effects of alliance component (agreement vs. bond), rater perspective (client vs. therapist) and timing (early vs. late therapy) on the alliance-outcome relationship. Individuals with an anxiety disorder enrolled into transdiagnostic group CBT were studied, with n = 78 at early therapy and n = 57 at late therapy. Results showed that greater client-rated agreement significantly predicted improved post-treatment outcomes throughout the course of therapy, while stronger client-rated bond in late therapy predicted reduced treatment gains. In contrast, therapist perceptions of agreement and bond were not associated with post-treatment outcomes at any point in therapy. Client-reported group cohesion also was not associated with additional variance in outcome after accounting for client-rated alliance. Overall, the findings highlight the importance of prioritizing the client's perception of the client-therapist relationship in CBT for anxiety disorders, as well as distinguishing the effects of component, rater and timing in future process-outcome studies.

Molecular evolution of candidate genes involved in post-mating-prezygotic reproductive isolation.

Traits involved in post-copulatory interactions between the sexes may evolve rapidly as a result of sexual selection and/or sexual conflict, leading to post-mating-prezygotic (PMPZ) reproductive isolating barriers between diverging lineages. Although the importance of PMPZ isolation is recognized, the molecular basis of such incompatibilities is not well understood. Here, we investigate molecular evolution of a subset of Drosophila mojavensis and Drosophila arizonae reproductive tract genes. These include genes that are transcriptionally regulated by conspecific mating in females, many of which are misregulated in heterospecific crosses, and a set of male genes whose transcripts are transferred to females during mating. As a group, misregulated female genes are not more divergent and do not appear to evolve under different selection pressures than other female reproductive genes. Male transferred genes evolve at a higher rate than testis-expressed genes, and at a similar rate compared to accessory gland protein genes, which are known to evolve rapidly. Four of the individual male transferred genes show patterns of divergent positive selection between D. mojavensis and D. arizonae. Three of the four genes belong to the sperm-coating protein-like family, including an ortholog of antares, which influences female fertility and receptivity in Drosophila melanogaster. Synthesis of these molecular evolutionary analyses with transcriptomics and predicted functional information makes these genes candidates for involvement in PMPZ reproductive incompatibilities between D. mojavensis and D. arizonae.

Development and stability of bacteriocin resistance in Campylobacter spp.

AIMS: Several bacteriocins (BCNs) that were identified from chicken commensal bacteria dramatically reduced Campylobacter colonization in poultry and are being directed toward on-farm control of this important foodborne human pathogen. A recent study has shown that BCN resistance in Campylobacter jejuni is very difficult to develop in vitro. In this study, in vivo development and stability of BCN resistance in Campylobacter was examined. METHODS AND RESULTS: Chickens infected with Camp. jejuni NCTC 11168 were treated with BCN E-760 at the dose of 5 mg kg(-1) body weight day(-1) via oral gavages for three consecutive days, which selected BCN-resistant (BCN(r)) mutants in the treated birds. However, all the in vivo-selected mutants only displayed low levels of resistance to BCN (MIC = 2-8 mg l(-1)) when compared to parent strain (MIC = 0.5 mg l(-1)). Inactivation of CmeABC efflux pump of the BCN(r) mutants led to increased susceptibility to BCN (8-32 fold MIC reduction). Three different BCN(r) Campylobacter strains (in vitro- or in vivo-derived) were examined for the stability of BCN resistance using both in vitro and in vivo systems. The low level of BCN resistance in these strains was not stable in vitro or in vivo in the absence of BCN selection pressure. CONCLUSIONS: Usage of BCN E-760 only selected low-level BCN(r) Camp. jejuni mutants in vivo, and the low-level BCN resistance was not stable in vitro and in vivo. SIGNIFICANCE AND IMPACT OF THE STUDY: The study provides helpful information for risk assessment of the future practical application of the anti-Campylobacter BCNs in animals.

Elements of the therapeutic relationship in CBT for anxiety disorders: A systematic review.

To optimise the effects of cognitive behaviour therapy (CBT) for anxiety disorders, research has increasingly focussed on understanding mechanisms of change. Specifically, the therapeutic relationship has been identified as a potential "active ingredient" of therapy. The evidence for the effects of eleven elements of the therapeutic relationship (alliance, collaboration, goal consensus, group cohesion, empathy, positive regard, feedback, emotional expression, outcome expectations, treatment credibility, alliance rupture-repair) on treatment outcomes in CBT for anxiety disorders was systematically reviewed. Fifty unique studies were included, and findings were qualitatively reviewed and summarised. Results revealed consistent and sizeable evidence for the cohesion-outcome and expectation-outcome relationships. There was emerging evidence for the effects of collaboration, empathy, and alliance rupture-repair on outcomes. However, the evidence for goal consensus and credibility on outcomes was limited. Notably, review of the alliance literature revealed substantial inconsistencies across studies. No studies were identified for positive regard, feedback, and emotional expression. Overall, further research is needed to clarify the role of the therapeutic relationship in CBT for anxiety disorders. These findings will contribute to the conceptual integration of therapeutic relationship constructs in cognitive behavioural models, and help to improve treatments and outcomes for individuals.

Activation of membrane-associated procaspase-3 is regulated by Bcl-2.

The mechanism by which membrane-bound Bcl-2 inhibits the activation of cytoplasmic procaspases is unknown. Here we characterize an intracellular, membrane-associated form of procaspase-3 whose activation is controlled by Bcl-2. Heavy membranes isolated from control cells contained a spontaneously activatable caspase-3 zymogen. In contrast, in Bcl-2 overexpressing cells, although the caspase-3 zymogen was still associated with heavy membranes, its spontaneous activation was blocked. However, Bcl-2 expression had little effect on the levels of cytoplasmic caspase activity in unstimulated cells. Furthermore, the membrane-associated caspase-3 differed from cytosolic caspase-3 in its responsiveness to activation by exogenous cytochrome c. Our results demonstrate that intracellular membranes can generate active caspase-3 by a Bcl-2-inhibitable mechanism, and that control of caspase activation in membranes is distinct from that observed in the cytoplasm. These data suggest that Bcl-2 may control cytoplasmic events in part by blocking the activation of membrane-associated procaspases.

Dexamethasone protects organ of corti explants against tumor necrosis factor-alpha-induced loss of auditory hair cells and alters the expression levels of apoptosis-related genes.

OBJECTIVE: Determine the molecular mechanism(s) behind tumor necrosis factor-alpha (TNFalpha)-induced loss of auditory hair cells and the ability of dexamethasone base (DXMb) to protect against TNFalpha ototoxicity. METHODS: Hair cell counts: Three-day-old rat organ of Corti explants were cultured under three different conditions: 1) untreated-control; 2) TNFalpha (2 mug/ml); and 3) TNFalpha (2 mug/ml)+DXMb (70 mug/ml) for 4 days, fixed, and stained with FITC-phalloidin. Hair cells were counted in the basal and middle turns. Gene expression: total RNA was extracted from the three different groups of explants at 0, 12, 24 and 48 h. Using quantitative real-time RT-PCR, mRNAs were transcribed into cDNAs and amplification was performed using primers for rat ss-actin (housekeeping gene), TNFR1, Bcl-2, Bax, and Bcl-xl. RESULTS: DXMb protected explant hair cells from TNFalpha-induced loss. Bax gene expression was greater in TNFalpha-exposed explants compared with TNFalpha+DXMb-treated explants at 48 h (P=0.023), confirmed by the increase in the Bax/Bcl-2 ratio at 48 h (P<0.001). These results correlated with increased TNFR1 expression at 24 h (P=0.038). DXMb otoprotection in TNFalpha-exposed cultures was accompanied by an up-regulation of Bcl-xl at both the 24 (P<0.001) and 48 h time points (P=0.030) and up-regulation of Bcl-2 expression at 24 h (P=0.018). DXMb treatment also prevented increases in the expression levels of Bax, TNFR1, and the Bax/Bcl-2 ratio that occurred in untreated TNFalpha-exposed explants. CONCLUSIONS: TNFalpha's ototoxicity may be mediated through an up-regulation of Bax and TNFR1 expression as well as an increase in the Bax/Bcl-2 ratio. DXMb protects the organ of Corti against TNFalpha ototoxicity by up-regulating Bcl-2 and Bcl-xl expression and by inhibiting TNFalpha-induced increases in Bax, TNFR1, and the Bax/Bcl-2 ratio. These results support the use of local dexamethasone treatment to conserve hearing following a trauma.

Use of sequence analysis of the VP4 gene to classify recent Vietnamese rotavirus isolates.

Twenty-eight strains of P(8), four of P(4) and one of P(19) rotavirus, isolated in Ho Chi Minh City, Vietnam, during 2002-2003, were investigated by sequence analysis of the VP4 gene. Seven of the 28 P(8) rotavirus VP4 sequences clustered in the P(8)-3 lineage, or the rare, so-called OP354-like lineage. Amino-acid sequence comparison revealed that Vietnamese P(8)-3 rotaviruses were generally very similar to Malawian strains, including the prototype OP354 strain. The numerical severity scores of diarrhoeal disease caused by the Vietnamese P(8)-3 rotaviruses were statistically higher than those of diarrhoeal disease caused by rotaviruses in the more common P(8)-2 lineage. Sequence and phylogenetic analysis of the VP4 gene of a Vietnamese G9P(19) rotavirus isolate showed a high degree of homology with the cognate genes of other human and porcine rotaviruses, including the prototype 4F strain.

Purified Inactivated Zika Vaccine Candidates Afford Protection against Lethal Challenge in Mice.

In response to the 2016 global public health emergency of international concern announced by the World Health Organization surrounding Zika virus (ZIKV) outbreaks, we developed a purified inactivated Zika virus vaccine (PIZV) candidate from ZIKV strain PRVABC59, isolated during the outbreak in 2015. The virus isolate was plaque purified, creating six sub-isolated virus stocks, two of which were selected to generate PIZV candidates for preclinical immunogenicity and efficacy evaluation in mice. The alum-adjuvanted PIZV candidates were highly immunogenic in both CD-1 and AG129 mice after a 2-dose immunization. Further, AG129 mice receiving 2 doses of PIZV formulated with alum were fully protected against lethal ZIKV challenge and mouse immune sera elicited by the PIZV candidates were capable of neutralizing ZIKVs of both African and Asian genetic lineages in vitro. Additionally, passive immunization of naïve mice with ZIKV-immune serum showed strong positive correlation between neutralizing ZIKV antibody (NAb) titers and protection against lethal challenge. This study supported advancement of the PIZV candidate toward clinical development.

Contrast-enhanced ultrasound in gallbladder disease: a pictorial review.

With its excellent contrast and spatial resolution, and the ability to image in real-time, ultrasound is the main imaging modality for assessing the gallbladder (GB). The application of contrast-enhanced ultrasound (CEUS) of the GB is now increasingly recognized as a useful addition to ultrasound and other cross-sectional imaging in the assessment of neoplastic and non-neoplastic GB disease. With the ability to image microcirculation and optimal contrast resolution, CEUS allows high-quality delineation in real-time, allowing for increased diagnostic confidence. In addition, ultrasound contrast agents have a favorable safety profile and can be used if CT or MR contrast agents are contraindicated or undesired. In this review, the CEUS appearances of a range of GB diseases encountered are presented, including adenomyomatosis, polyps, carcinoma, sludge, and cholecystitis with mural ulceration or perforation.

A comparison of independent component analysis algorithms and measures to discriminate between EEG and artifact components.

Independent Component Analysis (ICA) is a powerful statistical tool capable of separating multivariate scalp electrical signals into their additive independent or source components, specifically EEG or electroencephalogram and artifacts. Although ICA is a widely accepted EEG signal processing technique, classification of the recovered independent components (ICs) is still flawed, as current practice still requires subjective human decisions. Here we build on the results from Fitzgibbon et al. [1] to compare three measures and three ICA algorithms. Using EEG data acquired during neuromuscular paralysis, we tested the ability of the measures (spectral slope, peripherality and spatial smoothness) and algorithms (FastICA, Infomax and JADE) to identify components containing EMG. Spatial smoothness showed differentiation between paralysis and pre-paralysis ICs comparable to spectral slope, whereas peripherality showed less differentiation. A combination of the measures showed better differentiation than any measure alone. Furthermore, FastICA provided the best discrimination between muscle-free and muscle-contaminated recordings in the shortest time, suggesting it may be the most suited to EEG applications of the considered algorithms. Spatial smoothness results suggest that a significant number of ICs are mixed, i.e. contain signals from more than one biological source, and so the development of an ICA algorithm that is optimised to produce ICs that are easily classifiable is warranted.

Subtle overlapping deletions in the terminal region of chromosome 6q24.2-q26: three cases studied using FISH.

Interstitial deletions in the terminal region of chromosome 6 are rare. We describe three new cases with subtle interstitial deletions in the q24-q26 region of the long arm of chromosome 6. The karyotypes were analyzed at a 550 band level. Patient1 is a 9-month-old boy with an interstitial deletion, del(6)(q24.2q25.1), developmental delay, low birth weight, hypotonia, heart murmur, respiratory distress, craniofacial and genital anomalies. This is the first report of a case with deletion del(6)(q24.2q25.1). Patient 2 is a 17-year-old young man with an interstitial deletion del(6)(q25.1q25.3), developmental delay, short stature, mental retardation, autism, head, face, chest, hand and feet anomalies and a history of seizures. For the first time autism was described as a manifestation in 6q deletions. Patient 3 is baby boy with a de novo interstitial deletion, del(6)(q25.1q26), anomalies of the brain, genital organs, limbs and feet. This is the first report of a case with deletion, del(6)(q25.1q26). In all three patients, fluorescence in situ hybridization (FISH) using chromosome 6 painting probe ruled out an insertion. The ESR (6q25.1) and TBP (6q27) probes were used to confirm the breakpoints. Since TBP signal is present in all cases, it confirmed an interstitial deletion proximal to this probe. Patient 1 has a deletion of the ESR locus; Patient 2 and 3 have signals for the ESR locus on both chromosomes 6. Therefore the deletion in Patients 2 and 3 are between ESR and TBP loci distal to that of Patient 1. FISH validated the deletion breakpoints assessed by conventional cytogenetics.

Benign thyroid hyperplasia presenting as bilateral vocal cord paralysis. Complete remission following surgery.

A 52-year-old woman developed respiratory arrest on two separate occasions that required mechanical ventilation. Fiberoptic bronchoscopy demonstrated bilateral vocal cord paralysis, and a CT scan of the neck demonstrated a right neck mass. On surgical exploration, the mass was found extending from the thyroid gland and was identified as benign thyroid tissue. Thyroid hyperplasia should be considered in the differential diagnosis of bilateral vocal cord paralysis.

Constrictive pericarditis due to coccidiomycosis.

Coccidiomycosis is a fungal infection that rarely causes cardiac disease. Constrictive pericarditis in the setting of disseminated coccidiomycosis can be fatal, despite antifungal therapy and pericardiectomy. We report on a patient with constrictive pericarditis due to localized infection by Coccidioides immitis. The patient underwent successful surgical pericardiectomy and antifungal chemotherapy, and remains well 1 year later.

Physiologically based pharmacokinetic models: mathematical fundamentals and simulation implementations.

This review paper gives an overview of the building blocks of physiologically based pharmacokinetic (PBPK) models and their implementation using computer facilities. The approach focuses on the development of a PBPK model with the most important and appropriate limiting steps for the conditions and exposure scenarios under study. In this approach, the assumptions made in constructing the set of equations, as well as the fitting of variables to specific experimental results, need to be accounted for when making extrapolation to other conditions. A well-constructed PBPK model should account for all possible ranges of extrapolation from the development stages so that appropriate experimental studies and assumptions can be designed to handle the intended applications. Two common assumptions are revisited: the flow-limited assumption and the metabolic clearance using Michaelis-Menten kinetics assumption. Computer hardware and software requirements for implementing PBPK models are briefly reviewed.

Correlation between gadolinium-diethylenetriaminepentaacetic acid contrast enhancement and thallium-201 chloride uptake in pediatric brainstem glioma.

We previously showed that thallium-201 (201Tl) chloride is accumulated in over 75% of brain tumors, including brainstem gliomas. The imaging of 201Tl with single photon emission computed tomography (SPECT) may require an abnormal increase in permeability of tumor vessels to allow penetration of the blood-brain barrier. To test this hypothesis, we evaluated the correlation between gadolinium enhancement and the degree of 201Tl uptake on SPECT and the contributions of either gadolinium enhancement or 201Tl uptake to the prognosis in children with brainstem gliomas. Forty-two sets of paired SPECT scans and magnetic resonance imaging (MRI) scans were obtained longitudinally in 13 cases. Altogether, 31 of 42 pairs (74%) of scans showed concordance between the presence of gadolinium enhancement and 201Tl uptake. There were no cases that demonstrated 201Tl uptake but lacked gadolinium enhancement. The results indicate that 201Tl SPECT is of value primarily when brainstem tumors have vessels that are demonstrably permeable to gadolinium, prior to or as a result of radiotherapy.

Gene therapy for neurologic disease: benchtop discoveries to bedside applications. 1. The bench.

The overall goal of this review is to provide the pediatric neurologist with a theoretical foundation in gene therapy. Gene therapy became feasible in the early 1970s and the first transfer of a foreign gene into humans was approved by the NIH in 1989. Adenovirus, adeno-associated virus, herpes-simplex virus, retroviruses, and other vectors have been used to efficiently transduce genes into cells in vitro and in vivo. We discuss laboratory experiments that have provided a strong scientific rationale for implementing human clinical trials of gene therapy for neurologic malignancy. The development of viral and nonviral vectors that mediate efficient gene insertion into human cells has created the prospect of using gene therapy for cancer or brain disease. The NIH has approved more than 100 gene therapy protocols since 1989. However, the field will require more research on gene delivery systems before gene therapy becomes an established therapeutic strategy for an array of central nervous system diseases.

Gene therapy for neurologic disease: benchtop discoveries to bedside applications. 2. The bedside.

Important advances in basic research have made it possible to examine the safety, toxicity, and efficacy of gene therapy in humans for over 5 years. The development of sophisticated gene delivery systems has resulted in approval by the Recombinant DNA Advisory Committee (RAC) of 125 gene therapy or gene marking studies. One of the primary applications of current retroviral-mediated gene insertion technology has been for malignant brain tumors. Studies are therefore underway to examine the efficacy of "suicide" gene therapy in children with recurrent brain tumors and adults with newly diagnosed or recurrent gliomas. Since a high proportion of genetic disorders produce neurologic dysfunction, gene therapy is likely to impact the management of neurologic disease in the foreseeable future. Patients with human immunodeficiency virus (HIV), Gaucher's disease, and Hunter syndrome are now enrolled in gene therapy trials. It will be challenging for the child neurologist to stay abreast of rapid developments in the field of gene therapy. By participating in the design and implementation of clinical trials in gene therapy, the neurologist may reduce the intense toll that several neurologic diseases take on children and their families.

"Joubert syndrome" revisited: key ocular motor signs with magnetic resonance imaging correlation.

Joubert syndrome is characterized by episodic hyperpnea and apnea, developmental delay, hypotonia, truncal ataxia, ophthalmologic abnormalities, and vermian dysgenesis. We studied 15 patients with the diagnosis of Joubert syndrome to (1) more fully define the syndrome's clinical features, and (2) correlate the clinical features with magnetic resonance imaging (MRI) findings. Eight of 15 patients had a history of episodic hyperpnea and apnea. All patients had developmental delay and hypotonia. Of the 13 patients receiving detailed neuro-ophthalmologic evaluations, three had optic nerve dysplasia, pendular nystagmus, and gaze-holding nystagmus. All 13 patients had a normal vestibulo-ocular reflex based on head thrust, but had absent to poor ability to cancel the vestibulo-ocular reflex horizontally and vertically. Twelve of 13 patients had impaired smooth pursuit. Twelve of 13 patients had defects in initiation of saccades and quick phases. Two of the most consistent radiologic features were absent or hypoplastic posterior cerebellar vermis, and deformed midbrain and pontomesencephalic junction, which based on ocular motor physiology correlate with the vestibulo-ocular reflex cancellation/ pursuit defect and saccade initiation defect, respectively. As a result of midbrain, vermian, and superior cerebellar peduncle abnormalities, axial neuroimaging showed a unique "molar tooth" appearance of these structures. These results indicate that Joubert syndrome results from maldevelopment of the midbrain and cerebellar vermis, producing a pathognomonic sign on MRI.