MHC class I evolution; from Northern pike to salmonids.

BACKGROUND: Salmonids are of major importance both as farmed and wild animals. With the changing environment comes changes in pathogenic pressures so understanding the immune system of all salmonid species is of essence. Major histocompatibility complex (MHC) genes are key players in the adaptive immune system signalling infection to responding T-cells populations. Classical MHC class I (MHCI) genes, defined by high polymorphism, broad expression patterns and peptide binding ability, have a key role in inducing immunity. In salmonids, the fourth whole genome duplication that occurred 94 million years ago has provided salmonids with duplicate MHCI regions, while Northern Pike, a basal sister clade to salmonids, represent a species which has not experienced this whole genome duplication. RESULTS: Comparing the gene organization and evolution of MHC class I gene sequences in Northern pike versus salmonids displays a complex picture of how many of these genes evolved. Regional salmonid Ia and Ib Z lineage gene duplicates are not orthologs to the Northern pike Z lineage sequences. Instead, salmonids have experienced unique gene duplications in both duplicate regions as well as in the Salmo and Oncorhynchus branch. Species-specific gene duplications are even more pronounced for some L lineage genes. CONCLUSIONS: Although both Northern pike as well as salmonids have expanded their U and Z lineage genes, these gene duplications occurred separately in pike and in salmonids. However, the similarity between these duplications suggest the transposable machinery was present in a common ancestor. The salmonid MHCIa and MHCIb regions were formed during the 94 MYA since the split from pike and before the Oncorhynchus and Salmo branch separated. As seen in tetrapods, the non-classical U lineage genes are diversified duplicates of their classical counterpart. One MHCI lineage, the L lineage, experienced massive species-specific gene duplications after Oncorhynchus and Salmo split approximately 25 MYA. Based on what we currently know about L lineage genes, this large variation in number of L lineage genes also signals a large functional diversity in salmonids.

Topical dura mater application of CFA induces enhanced expression of c-fos and glutamate in rat trigeminal nucleus caudalis: attenuated by KYNA derivate (SZR72).

BACKGROUND: Migraine is a debilitating neurological disorder where trigeminovascular activation plays a key role. We have previously reported that local application of Complete Freund's Adjuvant (CFA) onto the dura mater caused activation in rat trigeminal ganglion (TG) which was abolished by a systemic administration of kynurenic acid (KYNA) derivate (SZR72). Here, we hypothesize that this activation may extend to the trigeminal complex in the brainstem and is attenuated by treatment with SZR72. METHODS: Activation in the trigeminal nucleus caudalis (TNC) and the trigeminal tract (Sp5) was achieved by application of CFA onto the dural parietal surface. SZR72 was given intraperitoneally (i.p.), one dose prior CFA deposition and repeatedly daily for 7 days. Immunohistochemical studies were performed for mapping glutamate, c-fos, PACAP, substance P, IL-6, IL-1ß and TNFα in the TNC/Sp5 and other regions of the brainstem and at the C1-C2 regions of the spinal cord. RESULTS: We found that CFA increased c-fos and glutamate immunoreactivity in TNC and C1-C2 neurons. This effect was mitigated by SZR72. PACAP positive fibers were detected in the fasciculus cuneatus and gracilis. Substance P, TNFα, IL-6 and IL-1ß immunopositivity were detected in fibers of Sp5 and neither of these molecules showed any change in immunoreactivity following CFA administration. CONCLUSION: This is the first study demonstrating that dural application of CFA increases the expression of c-fos and glutamate in TNC neurons. Treatment with the KYNA analogue prevented this expression.

KYNA analogue SZR72 modifies CFA-induced dural inflammation- regarding expression of pERK1/2 and IL-1ß in the rat trigeminal ganglion.

BACKGROUND: Neurogenic inflammation has for decades been considered an important part of migraine pathophysiology. In the present study, we asked the question if administration of a novel kynurenic acid analogue (SZR72), precursor of an excitotoxin antagonist and anti-inflammatory substance, can modify the neurogenic inflammatory response in the trigeminal ganglion. METHODS: Inflammation in the trigeminal ganglion was induced by local dural application of Complete Freunds Adjuvant (CFA). Levels of phosphorylated MAP kinase pERK1/2 and IL-1ß expression in V1 region of the trigeminal ganglion were investigated using immunohistochemistry and Western blot. FINDINGS: Pretreatment with one dose of SZR72 abolished the CFA-induced pERK1/2 and IL-1ß activation in the trigeminal ganglion. No significant change was noted in case of repeated treatment with SZR72 as compared to a single dose. CONCLUSIONS: This is the first study that demonstrates that one dose of KYNA analog before application of CFA can give anti-inflammatory response in a model of trigeminal activation, opening a new line for further investigations regarding possible effects of KYNA derivates.

Dural administration of inflammatory soup or Complete Freund's Adjuvant induces activation and inflammatory response in the rat trigeminal ganglion.

BACKGROUND: Migraine is a painful disorder with a huge impact on individual and public health. We hypothesize that migraine pain originates from a central mechanism that results secondarily in hypersensitivity in peripheral afferents associated with the cerebral and cranial blood vessels. It has previously been shown that application of inflammatory or algesic substances onto the dura mater or chemical stimulation of the dural receptive fields causes hypersensitivity to mechanical and thermal stimulation together with direct activation of the TG. We asked whether local inflammation of dura mater induces inflammatory activation in the trigeminal ganglion. METHODS: We performed topical administration of inflammatory soup (IS) or Complete Freund's Adjuvant (CFA) onto an exposed area of the rat dura mater in vivo for 20 min. The window was closed and the rats were sacrificed after 4 h and up to 7 days. Myography was performed on middle meningeal arteries. The trigeminal ganglia were removed and processed for immunohistochemistry or Western blot. RESULTS: Both CFA and IS induced enhanced expression of pERK1/2, IL-1ß and CGRP in the trigeminal ganglia. The pERK1/2 immunoreactivity was mainly seen in the satellite glial cells, while IL-1ß reactivity was observed in the neuronal cytoplasm, close to the cell membrane, seemingly as sign of neuro-glial interaction. The CGRP expression in the neurons and nerve fibres was enhanced after the application of either inflammatory agent. Myography resulted in a strong vasoconstrictor response to IS, but not to CFA. CONCLUSIONS: These results suggest that the application of IS or CFA onto the dura mater causes long-term activation of the TG and demonstrate the importance of the neuro-glial interaction in the activation of the trigeminovascular system.

Changes in muscle fiber density following a stroke.

OBJECTIVES: Previous studies have revealed a selective functional loss of the large, high-threshold motor units in the paretic muscles after lesion of the upper motor neuron. We set out to study the degree and the time course of the reorganization of the motor units following a stroke. METHODS: Examinations were performed on 59 patients with a unilateral ischemic stroke in the territory of the middle cerebral artery, and on 42 healthy controls. The duration of hemiparesis ranged from 2 weeks to 48 months. The fiber density (FD) in the abductor digiti minimi muscle was determined by means of single-fiber electromyography on both the hemiparetic and the unaffected side in the patients, and unilaterally in the control subjects. RESULTS: The FD was increased on the hemiparetic side relative to the unaffected side and the control group. This change correlated with the severity of the clinical signs. The FD increased during the first 10 months following the stroke and subsequently remained stable. CONCLUSIONS: The process of reinnervation in the muscles takes place in the acute phase after stroke. These changes are related to the severity of the symptoms. SIGNIFICANCE: Our findings suggest that trans-synaptic degeneration of the spinal motor neurons occurs shortly after the lesion of the upper motor neurons.

Large motor units are selectively affected following a stroke.

OBJECTIVE: Previous studies have revealed a loss of functioning motor units in stroke patients. However, it remained unclear whether the motor units are affected randomly or in some specific pattern. We assessed whether there is a selective loss of the large (high recruitment threshold) or the small (low recruitment threshold) motor units following a stroke. METHODS: Forty-five stroke patients and 40 healthy controls participated in the study. Macro-EMG was recorded from the abductor digiti minimi muscle at two levels of force output (low and high). The median macro motor unit potential (macro-MUP) amplitude on the paretic side was compared with those on the unaffected side and in the controls. RESULTS: In the control group and on the unaffected side, the macro-MUPs were significantly larger at the high force output than at the low one. However, on the paretic side the macro-MUPs at the high force output had the same amplitude as those recorded at the low force output. These changes correlated with the severity of the paresis. CONCLUSIONS: Following a stroke, there is a selective functional loss of the large, high-threshold motor units. These changes are related to the severity of the symptoms. SIGNIFICANCE: Our findings furnish further insight into the pathophysiology of the motor deficit following a stroke.

F wave measurements detecting changes in motor neuron excitability after ischaemic stroke.

OBJECTIVE: To study mechanisms that are involved in the hyperexcitability of spinal motor neurons in upper motor neuron lesion, by means of F-wave measurements. METHODS: F waves from 44 hospitalized stroke patients and 35 healthy controls were recorded from abductor pollicis brevis muscles in the course of two experiments: (1) single stimuli following high-intensity ipsilateral cutaneous conditioning were used to stimulate the median nerve; (2) paired stimuli were given to the median nerve at gradually increasing interstimulus intervals to assess recovery curves. Mean F-wave amplitudes elicited by the conditioning stimuli were compared with mean F-wave amplitudes elicited by the test stimuli on both the hemiparetic and the unaffected side. RESULTS: There was no reduction on the hemiparetic side of mean F-wave amplitudes elicited following high-intensity ipsilateral cutaneous stimulation. The recovery of mean F-wave amplitude was completed at longer interstimulus intervals on the hemiparetic side than on the unaffected side and in controls. A correlation was found between this delay of recovery and hemiparetic severity. CONCLUSIONS: The results of our study support the role of changes in lower motor neuron membrane excitability in the enhancement of F wave amplitudes after stroke.

Design and fabrication of ultrafine piezoelectric composites.

Making fine scale (< 20 microm) piezoelectric composites for high frequency (> 50 MHz) ultrasound transducers remains challenging. Interdigital phase bonding (IPhB), described in this paper, presents a new technique developed to make piezoelectric composites at the ultrafine scale using a conventional dicing saw. Using the IPhB technique, a composite structure with a pitch that is less than the dicing saw blade thickness can be created. The approach is flexible enough to make composites of different combination of pitch and volume ratio. Using a conventional dicing saw with a 50 microm thick blade, composite with a 25 microm pitch and a volume ratio of 61 percent are fabricated. Such a composite is suitable for fabrication of ultrasonic transducers and arrays with central frequencies of up to 85 MHz. Single element transducers working at central frequencies of 50-60 MHz were made of these composites as a mean to characterize the acoustic performance. Measurement results of the transducers show that the longitudinal electromechanical coupling coefficient is greater than 0.6 and that there are no noticeable lateral resonances in the frequency range of 55-150 MHz. Design criteria for fine scale elements are also discussed based on theoretical results from finite element analysis (FEA).

[Epstein-Barr virus genome positive lymphoepithelioma-like carcinoma of the stomach]. / A gyomor Epstein-Barr-vírusgenom pozitív lymphoepitheliomaszerú carcinomája.

EBV is associated with a high number of tumours and non-tumourous conditions. The rare lymphoepithelioma like carcinoma of the stomach,--just as similar tumours of foregut origin (thymus, lung, salivary gland)--are frequently EBV genom positive with the expression of only a few genes (EBV nuclear antigen 1, EBV encoded ribonucleoproteins/EBER/, latency I). On the basis of the clinicopathological analysis of two cases and literature data the authors point out the male predominance and the relatively favourable prognosis of the patients, furthermore the frequent cardial-subcardial localization of these tumours. Since the frequent non-lymphoepithelioma like stomach tumours,--adenocarcinomas,--show EBV genom positivity in about 1% of the cases, it is concluded that the characteristic lymphoepithelioma like histological pattern is not a sine qua non condition of EBV genom positivity. It may also be assumed, that the CD8 and TIA 1 cytotoxic lymphocytes are not virus but tumour cell specific, however not efficient, perhaps not activated. The low level of apoptotic tumour cells supports this assumption. In one of the cases a double tumour, a genom positive lymphoepithelioma like carcinoma and a genom negative adenocarcinoma, adjacent to each other was seen which speaks in favour of common carcinogenetic factors and shows that microscopic neighbourhood is not a necessary condition in viral association. The origin of the possible oncogenic effect of EBV in the absence of the transforming gene products latent membrane protein 1 and EBNA 2 in genom positive stomach carcinomas is uncertain. The significance of the presence in both cases of CD 5+ tumour cells is not clear, the study of further cases is indicated.

Cooperative oligonucleotides in purification of cycle sequencing products.

Nucleic acid hybridization is an essential component in many of today's standard molecular biology techniques. In a recent study, we investigated whether nucleic acid capture could be improved by taking advantage of stacking hybridization, which refers to the stabilizing effect that exists between oligonucleotides when they hybridize in a contiguous tandem fashion. Here, we describe a specific approach for purification of sequencing products using cooperative probes that hybridize to single-strand targets where one of the probes has been coupled to a magnetic bead. This approach has been developed for standard sequencing primers and has been applied to shotgun plasmid libraries. The cooperative probes have been designed to anneal within the common vector sequence and to avoid co-purification of nonextended sequencing primers and misprimed sequencing products. The reuse of magnetic beads, together with salt independent elution, makes the approach suitable for high-capacity capillary electrophoresis instruments.

Single element high frequency (<50 MHz) PZT sol gel composite ultrasound transducers.

A sol gel composite process has been used to produce lead zirconate titanate coatings in the thickness range of 3 to 100 microm on aluminum substrates. The complex permittivity (epsilon(33)(S )), elastic stiffness (c(33)(D)), and the piezoelectric constant (h(33)) of the coating and the complex elastic stiffness (c(33)(D)) of the substrate have been determined using impedance measurements and a commercially available software program [Piezoelectric Resonance Analysis Program PRAP 2.0, TASI Technical Software, Kingston, Ontario, Canada]. The complex components of the material parameters account for the losses within the film and the substrate. Sol gel composite films on aluminum have a dielectric constant of 220 with an imaginary component of 1% and an electromechanical coupling coefficient of up to 0.24 with an imaginary component of 3%. These films are applied to the fabrication of a high frequency transducers suitable for ultrasound biomicroscopy (UBM). By combining the sol gel composite material with existing transducer fabrication techniques, single-element focusing transducers have been produced that operate in the frequency range of 70 to 160 MHz. Devices have -6-dB bandwidths up to 52% and minimum insertion losses ranging from -47 to -58 dB. Real-time images of phantom materials and ex vivo biological samples are shown.

[Neurofibromatosis (von Recklinghausen disease) and carcinoid tumor in Vater's ampulla]. / Neurofibromatosis (von Recklinghausen-betegség) és carcinoid tumor a Vater-papillában.

The localization of the carcinoid tumors of the gastrointestinal tract--as a part of the APUD system--is extremely seldom at the region of ampulla of Vater. A case of congenital neurofibromatosis is reported in a 49-year-old male patient, with a carcinoid tumor of the ampulla of Vater in the background of long-standing, atypical biliary complaints. Periampullary neoplasm of neural-crest origin are very rare, up to this time 73 cases of Vater papilla carcinoid tumors were described. Regarding the development of VP carcinoid in neurofibromatosis, our patient is the 21st reported case in the world literature, and the first one in Hungary. The authors review the literature, giving special attention to the risk of periampullary neoplasms in von Recklinghausen's disease, the need of accurate diagnosis and the correctly selected operative intervention.