Study of population genetic polymorphism and gene flow rate in Indian snow trout, Schizothorax richardsonii fish of Himalaya, India.

The genetic polymorphism and gene flow rate among the Indian snow trout fish population S. richadsonii from three different locations viz., Chirapani stream of Champawat district, Kosi and Gola river of Nainital district, Uttarakhand State, India were assessed by employing twenty numbers of Randomly Amplified Polymorphic DNA (RAPD) markers. The overall percent polymorphisms among these three populations were 14.76 with 6.56, 4.92 and 3.28 in Chirapani, Kosi and Gola river population, respectively. Chirapani population had higher proportion of polymorphic loci as compared to the Kosi and Gola. The higher value of genetic distance (0.1565) was obtained between Chirapani and Gola population and the lower value of genetic distance was observed between Chirapani and Kosi (0.1058) river population. The cluster analysis revealed that in the formation of two clusters, one consisted of Chirapani and Kosi and the other was Gola fish population. Gst estimates among these populations showed some extent of homogeneity with lower genetic differentiation rate between populations and further suggested that higher tolerance to mutation, as expected that RAPD bands, arose from both coding and non-coding DNA regions. The findings revealed that the rate of gene flow in three populations seemed very low i.e. highly conserved its genetic diversity in their natural waterbodies and indicative of little migration among populations (geographically isolated and not the possibilities man made interventions/introduction of similar kind of fish species). It is further concluded that the Chirapani, Kosi and Gola river populations of S. richardsonii were being conserved naturally in their habitat and the species actual genetic potential were being maintained (adaptation to local climatic conditions, reproduction, production traits and disease resistance trait etc) in their natural habitat.

Understanding the processes underpinning IMPlementing IMProved Asthma self-management as RouTine (IMP2ART) in primary care: study protocol for a process evaluation within a cluster randomised controlled implementation trial.

BACKGROUND: Providing supported self-management for people with asthma can reduce the burden on patients, health services and wider society. Implementation, however, remains poor in routine clinical practice. IMPlementing IMProved Asthma self-management as RouTine (IMP2ART) is a UK-wide cluster randomised implementation trial that aims to test the impact of a whole-systems implementation strategy, embedding supported asthma self-management in primary care compared with usual care. To maximise opportunities for sustainable implementation beyond the trial, it is necessary to understand how and why the IMP2ART trial achieved its clinical and implementation outcomes. METHODS: A mixed-methods process evaluation nested within the IMP2ART trial will be undertaken to understand how supported self-management was implemented (or not) by primary care practices, to aid interpretation of trial findings and to inform scaling up and sustainability. Data and analysis strategies have been informed by mid-range and programme-level theory. Quantitative data will be collected across all practices to describe practice context, IMP2ART delivery (including fidelity and adaption) and practice response. Case studies undertaken in three to six sites, supplemented by additional interviews with practice staff and stakeholders, will be undertaken to gain an in-depth understanding of the interaction of practice context, delivery, and response. Synthesis, informed by theory, will combine analyses of both qualitative and quantitative data. Finally, implications for the scale up of asthma self-management implementation strategies to other practices in the UK will be explored through workshops with stakeholders. DISCUSSION: This mixed-methods, theoretically informed, process evaluation seeks to provide insights into the delivery and response to a whole-systems approach to the implementation of supported self-management in asthma care in primary care. It is underway at a time of significant change in primary care in the UK. The methods have, therefore, been developed to be adaptable to this changing context and to capture the impact of these changes on the delivery and response to research and implementation processes.

Obstructive nephropathy secondary to AA-type amyloidosis in ankylosing spondylitis.

AA-type amyloidosis of the genitourinary tract is a rare phenomenon and few cases are described in the literature. We report a 42-year-old man with ankylosing spondylitis, who developed hematuria, bilateral hydronephrosis, and renal failure caused by AA amyloidosis.

Histological changes in human dental pulp following application of intrusive and extrusive orthodontic forces.

The aim of this study was to compare the effects of orthodontic extrusive and intrusive forces on histological changes of the human dental pulp. In this clinical trial, 52 sound upper first premolars from 26 patients scheduled for extraction for orthodontic reasons were selected. They were divided into 2 groups, based on the time intervals for histological evaluation (3 days and 3 weeks). In each group, 10 teeth received orthodontic extrusive forces, 10 teeth underwent intrusive forces, and 6 teeth served as controls. After each period, teeth were extracted and prepared for histological examination under light microscopy and some histological parameters were evaluated. The data were statistically analyzed by Kruskall-Wallis and Mann Whitney tests. Of the parameters evaluated, just vacuolization and disruption of the odontoblastic layer showed statistically significant differences between the control group and both of the experimental groups in each test period (P < 0.05). Additionally, there was no significant difference between 3-day and 3-week intervals in each experimental group, except for fibrosis in the extrusive group which significantly increased after 3 weeks of force application (P = 0.001). Histological pulp changes following extrusive and intrusive force applications for 3 days and 3 weeks show no difference from each other.

Identification, Prediction and Data Analysis of Noncoding RNAs: A Review.

BACKGROUND: Noncoding RNAs (ncRNAs) which play an important role in various cellular processes are important in medicine as well as in drug design strategies. Different studies have shown that ncRNAs are dis-regulated in cancer cells and play an important role in human tumorigenesis. Therefore, it is important to identify and predict such molecules by experimental and computational methods, respectively. However, to avoid expensive experimental methods, computational algorithms have been developed for accurately and fast prediction of ncRNAs. OBJECTIVE: The aim of this review was to introduce the experimental and computational methods to identify and predict ncRNAs structure. Also, we explained the ncRNA's roles in cellular processes and drugs design, briefly. METHOD: In this survey, we will introduce ncRNAs and their roles in biological and medicinal processes. Then, some important laboratory techniques will be studied to identify ncRNAs. Finally, the state-of-the-art models and algorithms will be introduced along with important tools and databases. RESULTS: The results showed that the integration of experimental and computational approaches improves to identify ncRNAs. Moreover, the high accurate databases, algorithms and tools were compared to predict the ncRNAs. CONCLUSION: ncRNAs prediction is an exciting research field, but there are different difficulties. It requires accurate and reliable algorithms and tools. Also, it should be mentioned that computational costs of such algorithm including running time and usage memory are very important. Finally, some suggestions were presented to improve computational methods of ncRNAs gene and structural prediction.

Nephrotic syndrome complicating chronic visceral leishmaniasis: re-emergence in patients with AIDS.

Leishmania infection may be associated with immunecomplex-mediated glomerular injury. Contrary to immune-competent individuals, leishmaniasis in HIV patients is a chronic, relapsing disease. Despite the increasing frequency of the Leishmania/ HIV co-infection, there is a paucity of information on the effects of such co-infection in the kidney. We present a patient with AIDS and refractory, relapsing visceral leishmaniasis who developed nephrotic syndrome associated with renal involvement by Leishmania in the absence of immunecomplex glomerular deposition. For the first time, the relapsing nature of renal injury in this context is documented.

Anti-angiogenic drug scheduling optimisation with application to colorectal cancer.

Bevacizumab (bvz) is a first choice anti-angiogenic drug in oncology and is primarily administered in combination with chemotherapy. It has been hypothesized that anti-angiogenic drugs enhance efficacy of cytotoxic drugs by "normalizing" abnormal tumor vessels and improving drug penetration. Nevertheless, the clinical relevance of this phenomenon is still unclear with several studies over recent years suggesting an opposing relationship. Herein, we sought to develop a new computational tool to interrogate anti-angiogenic drug scheduling with particular application in the setting of colorectal cancer (CRC). Specifically, we have employed a mathematical model of vascular tumour growth which interrogates the impact of anti-angiogenic treatment and chemotherapeutic treatment on tumour volume. Model predictions were validated using CRC xenografts which underwent treatment with a clinically relevant combinatorial anti-angiogenic regimen. Bayesian model selection revealed the most appropriate term for capturing the effect of treatments on the tumour size, and provided insights into a switch-like dependence of FOLFOX delivery on the tumour vasculature. Our experimental data and mathematical model suggest that delivering chemotherapy prior to bvz may be optimal in the colorectal cancer setting.

Author Correction: Anti-angiogenic drug scheduling optimisation with application to colorectal cancer.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

The microRNA-205-5p is correlated to metastatic potential of 21T series: A breast cancer progression model.

MicroRNA is a class of noncoding RNAs able to base pair with complementary messenger RNA sequences, inhibiting their expression. These regulatory molecules play important roles in key cellular processes including cell proliferation, differentiation and response to DNA damage; changes in miRNA expression are a common feature of human cancers. To gain insights into the mechanisms involved in breast cancer progression we conducted a microRNA global expression analysis on a 21T series of cell lines obtained from the same patient during different stages of breast cancer progression. These stages are represented by cell lines derived from normal epithelial (H16N2), atypical ductal hyperplasia (21PT), primary in situ ductal carcinoma (21NT) and pleural effusion of a lung metastasis (21MT-1 and 21MT-2). In a global microRNA expression analysis, miR-205-5p was the only miRNA to display an important downregulation in the metastatic cell lines (21MT-1; 21MT-2) when compared to the non-invasive cells (21PT and 21NT). The lower amounts of miR-205-5p found also correlated with high histological grades biopsies and with higher invasion rates in a Boyden chamber assay. This work pinpoints miR-205-5p as a potential player in breast tumor invasiveness.

Recurrent acute postinfectious glomerulonephritis.

Recurrent acute postinfectious glomerulonephritis is infrequent in childhood and exceptional in adults. The factors that determine recurrence are poorly understood. Selective IgA deficiency is characterized by an increased incidence of gastrointestinal and respiratory infections. The case of a 33-year-old man with a history of repetitive sinopulmonary infections and diagnosed with selective IgA deficiency is described. He suffered 2 episodes of postinfectious glomerulonephritis within a 15-year period. Selective IgA deficiency may have predisposed to the development of recurrent postinfectious glomerulonephritis

Heparin and the solubilization of asymmetric acetylcholinesterase.

Heparin solubilizes asymmetric acetylcholinesterase, from chick skeletal muscle and retina, as a 24 S complex which is quantitatively converted to conventional asymmetric molecular forms of the enzyme (A12 and A8, either class I or class II) upon exposure to high salt. The simultaneous presence of salt and heparin in the homogenization medium selectively prevents, however, the release of class II A-forms in both muscle and retina. Heparin may generally act by displacing native proteoglycans involved in the attachment of the enzyme tail to the extracellular matrix, or its neural equivalent, being in turn removed by salt to yield typical asymmetric enzyme forms. Heparin would also appear to displace some other molecules specifically involved in the EDTA-sensitive attachment of class II tailed forms, this effect being antagonized by salt.

Chondroitinases release acetylcholinesterase from chick skeletal muscle.

Bacterial chondroitinases (both ABC and AC types) release asymmetric and globular forms of AChE from chick skeletal muscle samples. Heparinases, however, including heparitinase I, fail to do so under different incubation conditions. These results do not support the direct implication of the heparin/heparan sulfate family of GAGs in the interaction of the different AChE molecular forms with the muscle ECM. GAGs of the chondroitin/dermatan sulfate group could however be involved, either directly or indirectly, in the attachment of the AChE collagen-like tail to the muscle basal lamina.

Guanine nucleotides protect against kainate toxicity in an ex vivo chick retinal preparation.

Ex vivo preparations of chick neural retina have been successfully used in the assessment of excitotoxicity and in the evaluation of the protective effects of glutamate antagonists. Using a variation of this approach, and measuring the acute and delayed toxic effects of kainate (KA) in terms of lactate dehydrogenase release, we have shown that guanine nucleotides behave as effective neuroprotecting agents. The anti-excitotoxic potency of guanine nucleotides (in the case of GMP and GDPbetaS it is about 100 times lower than that of DNQX, a powerful kainate antagonist) correlates well with their ability to displace KA from retinal KA receptors.

GABAergic modulation of acetylcholine release in cholinergic synaptosomes from Torpedo marmorata electric organ.

GABAA- and GABAB-receptor-specific agonists inhibit the depolarization-evoked release of acetylcholine in cholinergic synaptosomes from Torpedo electric organ. Over 60% of the release is inhibited by a 10(-4) M concentration of GABA itself. IC50s for muscimol and baclofen are 1.3 x 10(-4) and 2.2 x 10(-6) M, respectively. The effect of muscimol is totally blocked by the direct antagonist bicuculline methiodide, and also by the allosteric antagonists methyl 6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate, picrotoxinin and tert-butylbicyclo-orthobenzoate; the effect of baclofen is blocked by delta-aminovalerate. Furthermore, the inhibitory action of muscimol on acetylcholine release is substantially enhanced by flunitrazepam and pentobarbital. These results suggest the existence of typical GABAA and GABAB receptors in the presynaptic nerve terminals of the Torpedo electric organ regulating the liberation of acetylcholine and therefore the discharge of the electroplaques.

Characterization of binding sites for [(3)H]Gaba in Drosophila melanogaster CNS membranes.

Drosophila melanogaster membranes prepared from cephalic ganglia contain two apparently independent populations of saturable binding sites for [(3)H]GABA: the higher-affinity sites (K(d) = 23 nM; B(max = 0.97 pmol/mg)) are already labeled at 0 degrees C, while a lower-affinity population of binding sites (K(d = 258 nM); B(max = 8.51 pmol/mg)) can be additionally identified at 23 degrees C and higher temperatures. GABA and then muscimol, together with ?-aminovalerate, are very efficient displacers of the specific binding while the A-specific agonist isoguvacine, the A-specific antagonist bicuculline methiodide and the B-specific agonist baclofen have low to negligible effect. [(3)H]GABA binding is not modulated by allosteric effectors characteristic of the vertebrate A-site, except for an anomalous stimulation by DMCM. Both [(3)H]flunitrazepam and [(3)H]TBOB give "specific" binding values under different experimental conditions but, after kinetic analysis, the binding appears to be non-saturable in both cases. We conclude that there are marked differences between the Drosophila GABA receptor and the vertebrate GABA(A) receptor complex, although our results do not exclude the existence of an insect receptor complex, associated to a Cl(?) channel, with different structural and pharmacological properties.

The GABA(A) receptor complex in the developing chick optic tectum: characterization of [(3)H]muscimol binding sites.

As part of an ongoing study on the GABA(A) receptor complex in the developing chick optic tectum we describe some properties of the agonist site, as labeled by [(3)H]muscimol, including methodological, kinetic and pharmacological aspects. 16-day embryos and 10-day chicks have been selected as representative age points for the initial characterization of the receptor, prior to more detailed developmental studies. Our data indicate the existence, in both embryos and young birds, of a single class of statistically equivalent, high-affinity, saturable binding sites, with a dissociation constant (K(d)) of 80-90 nM in freeze-thawed/washed membranes, and about 8 nM in membranes additionally extracted with low concentrations of Triton X-100. Maximal densities of binding sites are nearly identical in both membrane preparations, ranging from ?2 to ?3 pmol/mg for the two age points considered. The pharmacological profiles suggest that avian receptors for [(3)H]muscimol are generally similar to the corresponding mammalian sites, behaving as typical bicuculline-sensitive, baclofen-insensitive type A GABA receptor sites. However, bicuculline and its derivatives are less efficient displacers of [(3)H]muscimol in detergent-extracted membrane preparations, being in all cases, as usually, much less effective displacers than GABA agonists. The effect of Triton X-100 on the muscimol site in the GABA(A) receptor, increasing the affinity for the radioligand by a factor of 10, and diminishing the efficiency of antagonists, is considered here in terms of structural changes in the receptor, induced by the action of the detergent on the membrane microenvironment.

The GABA(a) receptor complex in the developing chick optic tectum: Ontogeny of [(3)H]muscimol, [(3)H]flunitrazepam and [(35)S]TBPS binding sites.

The developmental profiles of the neurotransmitter recognition site, labeled by [(3)H]muscimol, and of the two main modulatory sites, labeled by [(3)H]flunitrazepam and [ (35)S ]t- butylbicyclophosphorothionate , respectively, within the GABA(A) receptor complex, have been determined in chick tectal lobes between embryonic day 8 and postnatal day 20. The consonance among the rates of appearance and accumulation of the three receptor sites in tectal membranes suggests a coordinated expression and assembly of the protein subunits involved in the spatial configuration of the complex and its three binding sites, although the existence of isolated muscimol binding subunits during early embryogenesis cannot be excluded at the present time. Furthermore, the total number of binding sites of each kind, per pair of lobes, is compatible with a 1/1/I stoichiometry. The GABA(A) receptor complex reaches a maximum of expression, relative to total membrane protein, immediately after hatching, suggesting that the tectal GABAergic system may be instrumental in damping the effects of sudden exposure to light of the chick visual system upon eye opening.

Two classes of collagen-tailed, asymmetric molecular forms of acetylcholinesterase in skeletal muscle: differential effects of denervation.

Skeletal muscles of different vertebrate species contain, as it is the case in other cholinergic tissues, two classes of collagen-tailed, asymmetric forms (A-forms) of acetylcholinesterase (AChE). Class I A-forms are readily brought into solution in the presence of high salt, while class II A-forms do additionally require a chelating agent, such as EDTA, for solubilization. All A-forms aggregate at low ionic strength but only class II A-forms are reaggregated by excess Ca(++), even in the presence of 1M NaCl. This Ca(++)-mediated aggregability of class II A-forms is slowly lost upon exposure to detergents such as Triton X-100. Although these two classes of AChE tailed forms seem to be present in endplate and non-endplate areas, and in both the extra- and intracellular compartments, class II A-forms are predominantly extracellular and endplate-specific, at least in the rat diaphragm. On the other hand, well-characterized fast- and slow-twitch muscles show no preference for either class of asymmetric AChE species. Upon denervation, class I A-forms are degraded faster and disappear earlier than their class II counterparts, which are still easily detectable 17 days after nerve section. Class I and class II AChE molecular species exist in similar relative proportions in many vertebrate muscles. Thus, collagen-tailed forms may be altogether more abundant, in skeletal muscle, than it was hitherto realized. It is expected that this further example of AChE polymorphism will contribute to a better understanding of cholinergic transmission in skeletal muscle and, more specially, of nerve-muscle interactions.

Kainic acid binding sites in the developing chick optic tectum.

A membrane fraction from embryonic or newborn chick tectum has been found to bind [(3)H]kainic acid in a specific, saturable manner. A similar membrane preparation from chick liver does not show any appreciable specific binding under the same experimental conditions. Specific [(3)H]kainic acid binding to tectal membranes is displaced by suitable concentrations of unlabelled kainic acid, l-glutamate and other excitatory amino acid analogues, both agonists and antagonists. The appearance and accumulation of kainic acid binding sites in chick optic tectum has been studied between embryonic day 8 and postnatal day 20. Although by hatching time a sizable amount of binding can be measured in tectal membranes, most of the receptor accumulation takes place after birth. Saturation kinetic studies of kainic acid binding, before and after birth, show one single binding mode with an apparent dissociation constant K (d) ? 160 nM and a binding capacity increasing from 0.83 pmol/mg of protein, at embryonic day 16, to 6.45 pmol/mg of protein at day 6 after birth. Changing levels of visual input (both deprivation and stimulation) modulate the postnatal rate of receptor accumulation, with a higher density of binding sites in visually deprived optic lobes and a lower density in stimulated lobes, as compared with tectal lobes in normally raised chicks.

Solubilization of collagen-tailed molecular forms of acetylcholinesterase from several brain areas in different vertebrate species.

The relative efficiency of a buffered medium containing a high salt concentration and EDTA as a means to solubilize collagen-tailed molecular forms of acetylcholinesterase has been examined in four brain areas of several species belonging to different vertebrate classes. This extraction procedure has proved successful in most cases, with the yield of tailed enzyme varying between less than 1 and 26% of the total tissue activity. The solubilization values are consistently higher in more primitive vertebrates than in mammals and, for a given species, are usually lower in the telencephalon than in other brain structures. Our results confirm that the vertebrate central nervous system contains collagen-tailed quaternary structural forms of acetylcholinesterase.