PCR–RFLP technique for species identification of molted feathers in six species of co-occurring Ardeids

Naturally molted feathers provide a valuable tissue resource for avian research. Here we present a convenient and accurate species identification method for the naturally molted white feathers of six co-occurring Ardeid species, Egretta eulophotes, E. garzetta, Ardea alba, Bubulcus ibis, Mesophoyx intermedia and Ardeola bacchus, based on the polymerase chain reaction–restriction fragment length polymorphism of the partial mitochondrial cytochrome c oxidase subunit I gene. The resulting species-specific restriction patterns are easily distinguished by agarose gel electrophoresis. In this study, 81 of 88 naturally molted feathers noninvasively collected from heronries and 139 plucked feathers were all successfully distinguished, while the 7 feathers that could not be successfully distinguished by this method did not provide enough amplified products. This newly developed method for species identification of molted feathers will be useful for further studies on the vulnerable Chinese Egret and the other five co-occurring Ardeid species in East Asia.     

A rapid method for species identification and interspecies introgression based on a PCR–RFLP: application to the closely related Neotropical toads Rhinella marina and R. schneideri (Anura: Bufonidae)

Toad species Rhinella marina and R. schneideri (Bufonidae, Anura) are widely distributed in Amazon and Cerrado/Caatinga, respectively. Recent data has hypothesized that these two toad species share a history of a massive introgressive hybridization. Therefore, the aim of the present work was the precise identification of organisms with signs of past introgression or actual ongoing hybridization. In this paper we conducted a method based on polymerase chain reaction–restriction fragment length polymorphism, through the use of the CfoI enzyme. The individuals of R. marina have two specific digested bands, which is not digested in R. schneideri, possessing the hybrids three bands. The method was very efficient and allowed fast identification of these species.     

Molecular analysis of NPC1 and NPC2 gene in 34 Niemann–Pick C Italian Patients: identification and structural modeling of novel mutations

Niemann–Pick C, the autosomal recessive neuro-visceral disease resulting from a failure of cholesterol trafficking within the endosomal–lysosomal pathway, is due to mutations in NPC1 or NPC2 genes. We characterized 34 unrelated patients including 32 patients with mutations in NPC1 gene and two patients in NPC2 gene. Overall, 33 distinct genotypes were encountered. Among the 21 unpublished NPC1 alleles, 15 were due to point mutations resulting in 13 codon replacements (p.C100S, p.P237L, p.R389L, p.L472H, p.Y634C, p.S636F, p.V780G, p.Q921P, p.Y1019C, p.R1077Q, p.L1102F, p.A1187V, and p.L1191F) and in two premature stop codons (p.R934X and p.Q447X); a new mutant carried two in cis mutations, p.[L648H;M1142T] and four other NPC1 alleles were small deletions/insertions leading both to frame shifts and premature protein truncations (p.C31WfsX26, p.F284LfsX26, p.E1188fsX54, and p.T1205NfsX53). Finally, the new intronic c.464-2A>C change at the 3′ acceptor splice site of intron 4 affected NPC1 messenger RNA processing. We also found a new NPC2 mutant caused by a change of the first codon (p.M1L). The novel missense mutations were further investigated by two bioinformatics approaches. Panther proein classification system computationally predicted the detrimental effect of all new missense mutations occurring at evolutionary conserved positions. The other bioinformatics approach was based on prediction of structural alterations induced by missense mutations on the NPC1 atomic models. The in silico analysis predicted protein malfunctioning and/or local folding alteration for most missense mutations. Moreover, the effects of the missense mutations (p.Y634C, p.S636F, p.L648H, and p.V780G) affecting the sterol-sensing domain (SSD) were evaluated by docking simulation between the atomic coordinates of SSD model and cholesterol.     

Functional and Molecular Characterization of a Novel Traumatic Peripheral Nerve–Muscle Injury Model

Traumatic injuries to human peripheral nerves are frequently associated with damage to nerve surrounding tissues including muscles and blood vessels. Currently, most rodent models of peripheral nerve injuries (e.g., facial or sciatic nerve) employ surgical nerve transection with scissors or scalpels. However, such an isolated surgical nerve injury only mildly damages neighboring tissues and weakly activates an immune response. In order to provide a rodent nerve injury model accounting for such nerve-associated tissue damage and immune cell activation, we developed a drop tower-based facial nerve trauma model in mice. We compare nerve regeneration in this novel peripheral nerve trauma model with the established surgical nerve injury along several parameters. These include gene expression, histological and functional facial motoneuron (FMN) regeneration, facial nerve degeneration, immune cell activation and muscle damage. Regeneration-associated genes (RAGs; e.g., Atf3) were strongly induced in FMNs subjected to traumatic and surgical injury. Regeneration of FMNs and functional recovery of whisker movement were faster in traumatic versus complete surgical injury, thus cutting down experimentation time. Wallerian degeneration of distal nerve stumps was readily observed in this novel trauma injury model. Importantly, drop tower-inflicted facial nerve injury resulted in muscle damage, activation of muscle satellite cell markers (PAX7) and pronounced infiltration of immune cells to the injury site only in this model but not upon surgical nerve transection. Thus, we provide a novel rodent PNS trauma model that can be easily adopted to other PNS nerves such as the sciatic nerve. Since this nerve trauma model replicates multiple tissue damage frequently encountered in clinical routine, it will be well suited to identify molecular and cellular mechanisms of PNS nerve repair in wild-type and genetically modified rodents.     

Rapid and reversible enhancement of blood–brain barrier permeability using lysophosphatidic acid

The present study characterizes the effects of lysophosphatidic acid (LPA) on blood–brain barrier (BBB) permeability focusing specifically on the time of onset, duration, and magnitude of LPA-induced changes in cerebrovascular permeability in the mouse using both magnetic resonance imaging (MRI) and near infrared fluorescence imaging (NIFR). Furthermore, potential application of LPA for enhanced drug delivery to the brain was also examined by measuring the brain accumulation of radiolabeled methotrexate. Exposure of primary cultured brain microvessel endothelial cells (BMECs) to LPA produced concentration-dependent increases in permeability that were completely abolished by clostridium toxin B. Administration of LPA disrupted BBB integrity and enhanced the permeability of small molecular weight marker gadolinium diethylenetriaminepentaacetate (Gd-DTPA) contrast agent, the large molecular weight permeability marker, IRdye800cwPEG, and the P-glycoprotein efflux transporter probe, Rhodamine 800 (R800). The increase in BBB permeability occurred within 3 minutes after LPA injection and barrier integrity was restored within 20 minutes. A decreased response to LPA on large macromolecule BBB permeability was observed after repeated administration. The administration of LPA also resulted in 20-fold enhancement of radiolabeled methotrexate in the brain. These studies indicate that administration of LPA in combination with therapeutic agents may increase drug delivery to the brain.     

Longitudinal study of reading and writing rehabilitation using a bigraph–biphone correspondence approach

Background: Treatments for sound‐blending ability in phonological dyslexia that train single grapheme–phoneme correspondences have had mixed success. A more recent approach to re‐establishing sound‐blending abilities is to train correspondences of bigraph–biphone units (CV + VC = CVC) (Berndt, Haendiges, Mitchum & Wayland, 1996 Berndt, R. S., Haendiges, A. N., Mitchum, C. C. and Wayland, S. C. 1996. An investigation of nonlexical reading impairments.. Cognitive Neuropsychology, 13(6): 763–801.  [Google Scholar]; Friedman & Lott, 2002 Friedman, R. B. and Lott, S. N. 2002. Successful blending in a phonological reading treatment for deep alexia.. Aphasiology, 16: 355–372. [Taylor & Francis Online], [Web of Science ®] , [Google Scholar]). This approach has proved beneficial thus far, although the reasons for its success are not yet fully understood.

Aims: The purpose of this longitudinal investigation was to use the bigraph–biphone segment‐blending approach to improve both reading and writing abilities in an individual with phonological dyslexia/dysgraphia. Re‐establishing this ability laid the foundation for continued treatment with longer words and phrases.

Methods & Procedures: A case study design combining reading and writing treatment was used in three treatment protocols. Initially, treatment focused on improving the participant's awareness of bigraph–biphone correspondences and sound‐blending abilities for one‐syllable nonwords. The successful completion of this protocol was followed by two treatments to extend these abilities to reading and writing two‐syllable words and eventually phrase‐length material.

Outcomes & Results: Gains were made in all treatment protocols. The participant progressed from an inability to read one‐syllable nonwords to reading and writing phrase‐length material.

Conclusions: This study provides further evidence that using bigraph–biphone correspondences to train sound‐blending abilities can improve both reading and writing abilities in cases of phonological dyslexia. Furthermore, the success of this treatment programme illustrates the benefit of a targeted treatment programme even 5 years post onset of aphasia, for reading and writing rehabilitation.     

Is bruxism associated with changes in neural pathways? A systematic review and meta-analysis of clinical studies using neurophysiological techniques

Brain Imaging and Behavior - This study aimed to systematically review the literature to identify clinical studies assessing neuroplasticity changes induced by or associated with bruxism or a...     

Characterization of 24 polymorphic microsatellite markers for Silene nutans,a gynodioecious–gynomonoecious species,and cross-species amplification in other Silene species

Silene nutans (Caryophyllaceae) is a rare, vulnerable plant species that exhibits gynodioecy, containing both female and hermaphroditic individuals in natural populations. We developed and characterized 24 novel polymorphic microsatellite markers from next-generation sequencing to gain insights into the mating system and population-genetic structure of this species. In 36 individuals from three populations, the number of alleles and expected heterozygosity ranged from 5 to 30 and from 0.156 to 0.903 respectively. Departures from panmixia were found for 58.33 % of the loci with a mean multilocus F _is estimate of 0.232, which is expected in a self-compatible species exhibiting a mixed-mating system. Cross-species amplification was examined among eight additional Silene species and was successful for 7–19 loci, depending on the taxa. Overall, these newly developed microsatellite markers exhibited a high level of polymorphism, which will facilitate paternity analyses and fine- and large-scale population-genetic studies.     

Olfactory identification deficits and associated response inhibition in obsessive-compulsive disorder: On the scent of the orbitofronto–striatal model

Olfactory identification ability implicates the integrity of the orbitofrontal cortex (OFC). The fronto–striatal circuits including the OFC have been involved in the neuropathology of Obsessive Compulsive Disorder (OCD). However, only a few studies have examined olfactory function in patients with OCD. The Brief Smell Identification Test (B-SIT) and tests from the Cambridge Neuropsychological Automated Battery (CANTAB) were administered to 25 patients with OCD and to 21 healthy matched controls. OCD patients showed a significant impairment in olfactory identification ability as well as widely distributed cognitive deficits in visual memory, executive functions, attention, and response inhibition. The degree of behavioural impairment on motor impulsivity (prolonged response inhibition Stop-Signal Reaction Time) strongly correlated with the B-SIT score. Our study is the first to indicate a shared OFC pathological neural substrate underlying olfactory identification impairment, impulsivity, and OCD. Deficits in visual memory, executive functions and attention further indicate that regions outside of the orbitofronto–striatal loop may be involved in this disorder. Such results may help delineate the clinical complexity of OCD and support more targeted investigations and interventions. In this regard, research on the potential diagnostic utility of olfactory identification deficits in the assessment of OCD would certainly be useful.     

Quantitative imaging of cerebral blood flow velocity and intracellular motility using dynamic light scattering–optical coherence tomography

This paper describes a novel optical method for label-free quantitative imaging of cerebral blood flow (CBF) and intracellular motility (IM) in the rodent cerebral cortex. This method is based on a technique that integrates dynamic light scattering (DLS) and optical coherence tomography (OCT), named DLS–OCT. The technique measures both the axial and transverse velocities of CBF, whereas conventional Doppler OCT measures only the axial one. In addition, the technique produces a three-dimensional map of the diffusion coefficient quantifying nontranslational motions. In the DLS–OCT diffusion map, we observed high-diffusion spots, whose locations highly correspond to neuronal cell bodies and whose diffusion coefficient agreed with that of the motion of intracellular organelles reported in vitro in the literature. Therefore, the present method has enabled, for the first time to our knowledge, label-free imaging of the diffusion-like motion of intracellular organelles in vivo. As an example application, we used the method to monitor CBF and IM during a brief ischemic stroke, where we observed an induced persistent reduction in IM despite the recovery of CBF after stroke. This result supports that the IM measured in this study represent the cellular energy metabolism-related active motion of intracellular organelles rather than free diffusion of intracellular macromolecules.     

A case report of two male siblings with autism and duplication of Xq13–q21, a region including three genes predisposing for autism

Autism spectrum disorder, severe behaviour problems and duplication of the Xq12 to Xq13 region have recently been described in three male relatives. To describe the psychiatric comorbidity and dysmorphic features, including craniosynostosis, of two male siblings with autism and duplication of the Xq13 to Xq21 region, and attempt to narrow down the number of duplicated genes proposed to be leading to global developmental delay and autism. We performed DNA sequencing of certain exons of the TWIST1 gene, the FGFR2 gene and the FGFR3 gene. We also performed microarray analysis of the DNA. In addition to autism, the two male siblings exhibited severe learning disability, self-injurious behaviour, temper tantrums and hyperactivity, and had no communicative language. Chromosomal analyses were normal. Neither of the two siblings showed mutations of the sequenced exons known to produce craniosynostosis. The microarray analysis detected an extra copy of a region on the long arm of chromosome X, chromosome band Xq13.1–q21.1. Comparison of our two cases with previously described patients allowed us to identify three genes predisposing for autism in the duplicated chromosomal region. Sagittal craniosynostosis is also a new finding linked to the duplication.     

Seroprevalence of anti-Toxoplasma gondii and anti-Borrelia species antibodies in patients with schizophrenia: a case–control study from western Turkey

Objectives. We examined IgG antibody seroprevalence and risk factors for anti-Toxoplasma gondii and anti-Borrelia sp. in schizophrenic patients. Methods. This case–control study included 30 schizophrenic patients and 60 healthy individuals. Serological analyses were identified by using ELISA technique. Results. In the case group the Toxoplasma seropositivity was 33.3% and Borrelia seropositivity was 13.3%, while in the control group the Toxoplasma positivity was 21.7% and Borrelia seropositivity was 15.0%. There was no significant difference with regard to seroprevalence between the groups (P = 0.232; P = 0.832, respectively). There was statistically significant difference between case and control groups related to hand and kitchen utensil hygiene after dealing with raw meat (P = 0.001). Conclusions. Our data showed the rate of Toxoplasma antibodies was higher in the case group, while the rate of Borrelia antibodies was higher in the control group. In both groups the high rates of seropositivity for Toxoplasma gondii and Borrelia sp. is thought to be due to neglect of personal hygiene. The present study also is the first to examine the association between Borrelia sp. and schizophrenia. Further studies are needed to determine whether there is an association between Borrelia sp. and schizophrenia or not.     

Measuring regularity of human postural sway using approximate entropy and sample entropy in patients with Ehlers–Danlos syndrome hypermobility type

Ligament laxity in Ehlers–Danlos syndrome hypermobility type (EDS-HT) patients can influence the intrinsic information about posture and movement and can have a negative effect on the appropriateness of postural reactions. Several measures have been proposed in literature to describe the planar migration of CoP over the base of support, and the most used in clinical field are the CoP excursions in antero-posterior and medio-lateral direction. In recent years a growing number of studies have been designed to explore the complexity of the COP trajectories during quiet standing. We assessed 13 adults with EDS-HT (EDSG) and 20 healthy adults (CG) during static posture, evaluating the CoP using time and frequency domain analysis and entropy analysis (SampEn and ApEn parameters). Higher values of CoP displacements in medio-lateral and anterior–posterior directions for EDSG than CG were found; no differences were observed in CoP frequency. The entropy analysis showed lower value for EDSG than CG, pointing out the needing of EDSG to concentrate more attention on postural control, loosing complexity and reflecting a less automatized postural control.     

Simultaneous measurement of glucose blood–brain transport constants and metabolic rate in rat brain using in-vivo 1H MRS

Cerebral glucose consumption and glucose transport across the blood–brain barrier are crucial to brain function since glucose is the major energy fuel for supporting intense electrophysiological activity associated with neuronal firing and signaling. Therefore, the development of noninvasive methods to measure the cerebral metabolic rate of glucose (CMR_glc) and glucose transport constants (K_T: half-saturation constant; T_max: maximum transport rate) are of importance for understanding glucose transport mechanism and neuroenergetics under various physiological and pathological conditions. In this study, a novel approach able to simultaneously measure CMR_glc, K_T, and T_max via monitoring the dynamic glucose concentration changes in the brain tissue using in-vivo¹H magnetic resonance spectroscopy (MRS) and in plasma after a brief glucose infusion was proposed and tested using an animal model. The values of CMR_glc, T_max, and K_T were determined to be 0.44±0.17 μmol/g per minute, 1.35±0.47 μmol/g per minute, and 13.4±6.8 mmol/L in the rat brain anesthetized with 2% isoflurane. The Monte-Carlo simulations suggest that the measurements of CMR_glc and T_max are more reliable than that of K_T. The overall results indicate that the new approach is robust and reliable for in-vivo measurements of both brain glucose metabolic rate and transport constants, and has potential for human application.     

Active PAI-1 as marker for venous thromboembolism: Case–control study using a comprehensive panel of PAI-1 and TAFI assays

Background

Both activated Thrombin Activatable Fibrinolysis Inhibitor (TAFI) and active Plasminogen Activator Inhibitor-1 (PAI-1) attenuate fibrinolysis and may therefore contribute to the pathophysiology of Venous ThromboEmbolism (VTE). Whether increased TAFI and/or PAI-1 concentrations are associated with VTE is unclear.

Objective

To study an association of impaired fibrinolysis and VTE using a comprehensive panel of in-house developed assays measuring intact TAFI, activation peptide of TAFI (AP-TAFI), PAI-1 antigen, endogenous PAI-1:t-PA complex (PAI-1:t-PA) and active PAI-1 levels in 102 VTE patients and in 113 healthy controls (HC).

Results

Active PAI-1 was significantly higher in VTE patients compared to HC (20.9 [9.6-37.8] ng/ml vs. 6.2 [3.5-9.7] ng/ml, respectively). Active PAI-1 was the best discriminator with an area under the ROC curve and 95% confidence interval (AUROC [95%CI]) of 0.84 [0.79-0.90] compared to 0.75 [0.68-0.72] for PAI-1:t-PA, 0.65 [0.58-0.73] for PAI-1 antigen, 0.62 [0.54-0.69] for AP-TAFI and 0.51 [0.44-0.59] for intact TAFI. Using ROC analysis, we defined an optimal cut-off of 12.8 ng/ml for active PAI-1, with corresponding sensitivity of 71 [61–79] % and specificity of 89 [82–94] %. A lack of association with the time between VTE event and sample collection or with the intake of anticoagulant treatment suggests that active PAI-1 levels are sustainable high in VTE patients.

Conclusions

This case–control study emphasizes the clinical importance of measuring active PAI-1 instead of PAI-1 antigen and identifies active PAI-1 as a potential marker of VTE. Prognostic studies will need to address the clinical significance of active PAI-1 as biomarker.