STING-associated vasculopathy with onset in infancy (SAVI) presenting with skin lesions.

STING-associated vasculopathy with onset in infancy (SAVI) is caused by pathogenic gain-of-function variants in the gene TMEM173 (also named stimulator of interferon genes, STING1). This report details the case of an 11-year-old girl with SAVI who presented with skin-limited symptoms and discusses the phenotype-genotype correlations of the TMEM173 variant present in our patient. Treatment of SAVI focuses on preventing the development or progression of organ damage by reducing systemic inflammation. We summarize the available treatments for this syndrome.

Gene therapy with mesenchymal stem cells expressing IFN-ß ameliorates neuroinflammation in experimental models of multiple sclerosis.

BACKGROUND AND PURPOSE: Recombinant IFN-ß is one of the first-line treatments in multiple sclerosis (MS), despite its lack of efficacy in some patients. In this context, mesenchymal stem cells (MSCs) represent a promising therapeutic alternative due to their immunomodulatory properties and multipotency. Moreover, by taking advantage of their pathotropism, these cells can be genetically modified to be used as carriers for delivering or secreting therapeutic drugs into injured tissues. Here, we report the therapeutic effect of systemic delivery of adipose-derived MSCs (AdMSCs), transduced with the IFN-ß gene, into mice with experimental autoimmune encephalomyelitis (EAE). EXPERIMENTAL APPROACH: Relapsing-remitting and chronic progressive EAE were induced in mice. Cells were injected i.v. Disease severity, inflammation and tissue damage were assessed clinically, by flow cytometry of spleens and histopathological evaluation of the CNS respectively. KEY RESULTS: Genetic engineering did not modify the biological characteristics of these AdMSCs (morphology, growth rate, immunophenotype and multipotency). Furthermore, the transduction of IFN-ß to AdMSCs maintained and, in some cases, enhanced the functional properties of AdMSCs by ameliorating the symptoms of MS in EAE models and by decreasing indications of peripheral and central neuro-inflammation. CONCLUSION AND IMPLICATIONS: Gene therapy was found to be more effective than cell therapy in ameliorating several clinical parameters in both EAE models, presumably due to the continuous expression of IFN-ß. Furthermore, it has significant advantages over AdMSC therapy, and also over systemic IFN-ß treatment, by providing long-term expression of the cytokine at therapeutic concentrations and reducing the frequency of injections, while minimizing dose-limiting side effects.

Recurrent lipoatrophic panniculitis of children.

BACKGROUND: Recurrent panniculitis in children with lipoatrophy has been loosely described and reported under different names, but has never been systematically evaluated by immunohistochemical stains. OBJECTIVE: To depict the profile of children with recurrent idiopathic panniculitis. METHODS: Study of clinical, histopathological and immunohistochemical features in five cases with recurrent idiopathic panniculitis. RESULTS: Five children with repeated attacks of painful subcutaneous nodules in association with fever, malaise and abdominal pain or arthralgia, with subsequent lipoatrophy were reviewed. In two patients, extensive involvement led to loss of the cutaneous fatty tissue. Laboratory abnormalities included increased acute phase reactants, leukocytosis with mild neutrophilia, microcytic anaemia and elevated liver enzymes. Histopathology showed lobar panniculitis without vasculitis and with a mixed infiltrate, composed of neutrophils, mononuclear cells, lymphocytes, macrophages and myeloid cells. Neutrophils and myeloid cells were more prominent in early lesions, whereas macrophages predominated in late stages, leading to lipophagia and lipoatrophy. Immunohistochemistry showed positive staining for myeloperoxidase around the necrotic adipocytes in early stages and CD68/PGM1 macrophages in late stages. Intense STAT1 staining was observed in the inflammatory infiltrate. All patients improved with methotrexate and corticosteroids. CONCLUSION: We present five cases of lobar panniculitis and lipoatrophy in childhood. The clinico-pathologic presentation shares features with other autoinflammatory diseases.

Recombinant soluble IFN receptor (sIFNAR2) exhibits intrinsic therapeutic efficacy in a murine model of Multiple Sclerosis.

Endogenous interferon beta (IFNß) is an important cytokine involved in several chronic inflammatory diseases, such as Multiple Sclerosis (MS). In spite of the numerous therapeutic approaches available for MS patients, the administration of recombinant IFNß continues being one of the first line treatment to these patients. The soluble form of IFNß receptor (sIFNAR2) could act as critical regulator of the endogenous and the systemically administered IFNß, but whether it functions as an agonist or antagonist of its ligand is not completely elucidated. Morover, the possible role of sIFNAR2 in autoimmune diseases like MS is still unknown and so far overlooked. Here we evaluated the efficacy of the combined therapy of IFNß and our recombinant protein analogous to human sIFNAR2 as a treatment in a chronic mice model of MS (CP-EAE). We also tested the effect of the sIFNAR2 administered as a monotherapy over these EAE-animals. The results showed that our recombinant sIFNAR2 protein potentiates the immunomodulatory effects of exogenous IFNß in CP-EAE by increasing the reduction of the induced inflammation and the tissue damage. Furthermore, we demonstrate for the first time that sIFNAR2 shows intrinsic properties by modulating the CP-EAE progression and the neuroinflammation processes related to this disease. Another intrinsic activity showed by sIFNAR2 is the inhibition of the T cells proliferation, which increase its potential as therapeutic molecule.

Olfactory system and demyelination.

Within the central nervous system, the olfactory system represents one of the most exciting scenarios since it presents relevant examples of long-life sustained neurogenesis and continuous axonal outgrowth from the olfactory epithelium with the subsequent plasticity phenomena in the olfactory bulb. The olfactory nerve is composed of nonmyelinated axons with interesting ontogenetic interpretations. However, the centripetal projections from the olfactory bulb are myelinated axons which project to more caudal areas along the lateral olfactory tract. In consequence, demyelination has not been considered as a possible cause of the olfactory symptoms in those diseases in which this sense is impaired. One prototypical example of an olfactory disease is Kallmann syndrome, in which different mutations give rise to combined anosmia and hypogonadotropic hypogonadism, together with different satellite symptoms. Anosmin-1 is the extracellular matrix glycoprotein altered in the X-linked form of this disease, which participates in cell adhesion and migration, and axonal outgrowth in the olfactory system and in other regions of the central nervous system. Recently, we have described a new patho-physiological role of this protein in the absence of spontaneous remyelination in multiple sclerosis. In the present review, we hypothesize about how both main and satellite neurological symptoms of Kallmann syndrome may be explained by alterations in the myelination. We revisit the relationship between the olfactory system and myelin highlighting that minor histological changes should not be forgotten as putative causes of olfactory malfunction.

Fenoterol e salbutamol via inalatória em cães: aspectos clínico, hemogasométrico e eletrocardiográfico / Fenoterol and salbutamol inhalation in dogs: clinical, haemogasometric and electrocardiographic aspects

Avaliaram-se, por meio de exame clínico, hemogasométrico e eletrocardiográfico, os efeitos do salbutamol e do fenoterol, administrados por via inalatória em cães. Doze cães foram distribuídos em dois grupos: os do grupo FE receberam fenoterol na dose de 2 gotas/5kg de peso vivo, diluídas em solução de cloreto de sódio 0,9 por cento por aparelho de inalação, e os do grupo SA receberam salbutamol pelo dosador de aerossol, na dose de 100mg. Foram avaliados: frequência cardíaca (FC), frequência respiratória (FR), temperatura retal (TR), pressão arterial sistólica (PAS), hemogasometria e eletrocardiograma antes e após 30min, duas horas e seis horas do uso dos fármacos. Discreta estimulação cardíaca ocorreu nos animais do grupo SA duas horas após sua administração em relação ao momento-controle, e tremores foram predominantes nestes animais. Diminuição da PAS e aumento da FR foram observados nos dois grupos, e não houve alteração significativa da onda T, da hemogasometria e do eletrocardiograma em ambos os grupos. O fenoterol provocou menor estimulação cardíaca e menos tremores comparado ao salbutamol, foi mais seguro e houve maior facilidade, menor custo e menor gasto de tempo na administração do salbutamol por inalador dosimetrado em relação ao fenoterol por nebulização.

Through physical examination, blood gas and the electrocardiographic effects of salbutamol and fenoterol, administered by inhalation in dogs was assessed. Twelve dogs were distributed into two groups: EF group animals received a fenoterol dose of 2 drops/5kg bodyweight, diluted in sodium chloride 0.9 percent inhalation device and animals in the SA group received salbutamol through aerosol feeder at a dose of 100µg. Rectal temperature (RT), heart rate (HR), respiratory rate (RR), systolic blood pressure (SBP), blood gas and electrocardiogram before and after 30min., 2h and 6 h after drug were evaluated. Mild cardiac stimulation occurred in SA group animals 2 hours after its administration compared to the control group, and tremors were predominant in these animals. A decrease in SBP and an increase in RR were observed in both animal groups and no significant alteration of the T wave in the electrocardiogram and blood gas analysis in both groups were observed. The fenoterol caused less cardiac pacing and shivering compared to salbutamol, which was more secure. However, there was more ease, lower cost and less time spent in the administration of salbutamol administered through metered-dose inhaler compared to fenoterol nebulization.

Guía de uso adecuado de modelos animales para el desarrollo de terapias en esclerosis múltiple / Guidelines on the appropriate use of animal models for developing therapies in multiple sclerosis

Introducción. La búsqueda de terapias efectivas para la esclerosis múltiple (EM) y la definición de ventanas terapéuticas apropiadas, así como el establecimiento de mejores biomarcadores diagnósticos y pronósticos, continúan siendo un reto para investigadores tanto básicos como clínicos. El desarrollo y el método de evaluación de los estudios preclínicos en modelos animales podrían subyacer al hecho de que terapias eficaces en modelos animales fracasen en su aplicación clínica. Objetivo. Unificar la metodología en la aplicación de los modelos experimentales para la EM mediante la elaboración, por parte de grupos españoles expertos pertenecientes a la Red Española de Esclerosis Múltiple, de una guía de recomendaciones para los estudios preclínicos. Desarrollo. Se ha realizado una valoración detallada de los modelos experimentales adecuados y su aplicación en función del objetivo perseguido, incorporando estándares y criterios de calidad imprescindibles en un estudio preclínico. Conclusiones. El éxito traslacional en el avance terapéutico de la EM conlleva la adquisición de compromisos metodológicos en los modelos experimentales, de manera que se optimice la bondad y adecuación del modelo al estudio perseguido. Las recomendaciones establecidas en esta guía podrían ayudar a generar datos preclínicos de utilidad en la práctica clínica (AU)

Introduction. The advance in the achievement of effective therapies for multiple sclerosis (MS), the definition of appropriate therapeutic windows and to establish better diagnostic and prognostic biomarkers have become a challenging task for both researchers and clinicians. Some pitfalls in clinical trials might be related to lack of adequacy of the preclinical studies in MS experimental animal models Aim. To standardize the methodological protocols of experimental models by developing a set of guidelines for preclinical studies by groups of experts from REEM (Spanish Network for MS). Development. A guide with recommendations for the application of MS models including a detailed assessment of appropriate experimental models taking into account the objective of the study that has been presented. Standards and quality criteria necessary in a preclinical study have been included. Conclusions. Standardized animal models of MS are essential to increase the success of the preclinical findings in order to transfer them to the clinical practice (AU)

Characterization of NADPH-diaphorase-positive glial cells of the tench optic nerve after axotomy.

NADPH-diaphorase (ND) positive cell types were characterized throughout the optic nerve of the tench in normal conditions and after optic nerve transection with survival periods of 1, 3, 7, 14, 30, 60, 120 and 180 days. Astrocytic markers (S100 and glutamine synthetase) and the microglial marker tomato lectin were employed. In the control prechiasmatic optic nerve two types (types I and II) of ND-positive glial cells appeared. All type I cells showed S100 immunoreactivity, whereas only a subpopulation of them were positive to glutamine synthetase. Type II cells only presented S100 immunoreactivity. In the control anterior optic tract, all ND-positive glial cells (type III) presented immunolabeling to S100 and glutamine synthetase. After transection, types I and II did not show any changes in the staining patterns for the glial markers when observed. Two new types of ND-positive glial cells (types IV and V) were observed after axotomy. All type IV cells were S100-immunopositive, and a subpopulation presented glutamine synthetase immunolabeling. Only a subpopulation of type V cells showed glutamine synthetase immunostaining. The presence of type IV or V cells in the lesioned optic nerve occurred simultaneously with significant decreases or absence of type I cells. These data suggest that type I and III cells are astrocytes and type II cells are oligodendrocytes. Types IV and V cells are the result of the activation of type I cells after optic nerve section. The polymorphism observed in ND-positive cells may reflect different cell functions during degenerative and regenerative processes.

El sistema cannabinoide en situaciones de neuroinflamación: perspectivas terapéuticas en la esclerosis múltiple / Cannabinoid system and neuroinflammation: therapeutic perspectives in multiple sclerosis

Introducción. El sistema endocannabinoide está constituidopor los receptores cannabinoides, los ligandos endógenos y loselementos enzimáticos implicados en su síntesis y degradación.Objetivo. Describir el estado actual de conocimiento sobre la funcióndel sistema como modulador de los procesos neuroinflamatoriosasociados con enfermedades crónicas como la esclerosis múltiple.Desarrollo. Los cannabinoides se sintetizan y se liberan en demanda y su producción aumenta en situaciones de neuroinflamacióny de daño neural. En este contexto, sus acciones en la microglía y enlos astrocitos se caracterizan por una disminución en la expresión demediadores inflamatorios y de citocinas proinflamatorias. Además,los cannabinoides pueden ejercer acciones neuroprotectoras a travésde diferentes tipos de mecanismos y en modelos experimentalesde esclerosis múltiple atenúan la sintomatología, disminuyen lainflamación y pueden favorecer la remielinización. Conclusiones. Eluso clínico de cannabinoides o agentes farmacológicos que incidenen el sistema endógeno cannabinoide durante la inflamación del sistemanervioso central y en la esclerosis múltiple está actualmentesometido a consideración y debate. El análisis detallado de los resultadosobtenidos en la última década ha permitido establecer que sonmúltiples los mecanismos de actuación de los cannabinoides en patologíasdel sistema nervioso central que cursan con inflamación crónicay ponen de manifiesto el interés del sistema cannabinoide comonueva herramienta terapéutica

Introduction. The endocannabinoid system consists of cannabinoid receptors, endogenous ligands and the enzymaticelements involved in their synthesis and breakdown. Aim. To report on currently held knowledge about the functioning of thesystem as a modulator of the neuroinflammatory processes associated with chronic diseases such as multiple sclerosis.Development. Cannabinoids are synthesised and released on demand and their production increases in times of neuroinflammationand neural damage. In this context then, their actions in the microglial cells and in the astrocytes arecharacterised by a lowered expression of inflammatory mediators and pro-inflammatory cytokines. Furthermore,cannabinoids can play a role as neuroprotectors by means of different types of mechanisms and, in experimental models ofmultiple sclerosis, they slow down the symptoms, reduce inflammation and can favour remyelination. Conclusions. Theclinical use of cannabinoids or pharmacological agents that affect the endogenous cannabinoid system during inflammationof the central nervous system and in multiple sclerosis is currently under consideration and subject to debate. Detailedanalysis of the results obtained over the past decade has made it possible to establish the existence of several mechanisms ofaction of cannabinoids in pathologies affecting the central nervous system that are accompanied by chronic inflammation.Likewise, they also clearly show that the cannabinoid system is an interesting proposal as a new therapeutic tool

[Cannabinoid system and neuroinflammation:therapeutic perspectives in multiple sclerosis]. / El sistema cannabinoide en situaciones de neuroinflamación: perspectivas terapéuticas en la esclerosis múltiple.

INTRODUCTION: The endocannabinoid system consists of cannabinoid receptors, endogenous ligands and the enzymatic elements involved in their synthesis and breakdown. AIM: To report on currently held knowledge about the functioning of the system as a modulator of the neuroinflammatory processes associated with chronic diseases such as multiple sclerosis. DEVELOPMENT: Cannabinoids are synthesised and released on demand and their production increases in times of neuroinflammation and neural damage. In this context then, their actions in the microglial cells and in the astrocytes are characterised by a lowered expression of inflammatory mediators and pro-inflammatory cytokines. Furthermore, cannabinoids can play a role as neuroprotectors by means of different types of mechanisms and, in experimental models of multiple sclerosis, they slow down the symptoms, reduce inflammation and can favour remyelination. CONCLUSIONS: The clinical use of cannabinoids or pharmacological agents that affect the endogenous cannabinoid system during inflammation of the central nervous system and in multiple sclerosis is currently under consideration and subject to debate. Detailed analysis of the results obtained over the past decade has made it possible to establish the existence of several mechanisms of action of cannabinoids in pathologies affecting the central nervous system that are accompanied by chronic inflammation. Likewise, they also clearly show that the cannabinoid system is an interesting proposal as a new therapeutic tool.

Centrosymmetric and pseudo-centrosymmetric structures refined as non-centrosymmetric.

The behaviour of the Flack parameter for centrosymmetric and pseudo-centrosymmetric crystal structures based on crystal structures published as being non-centrosymmetric is presented. It is confirmed for centrosymmetric structures that the value obtained for the Flack parameter is critically dependent on the Friedel coverage of the intensity data, approaching 0.5 for a coverage of 100% and sticking near the starting value for a coverage of 0%. For pseudo-centrosymmetric structures, even those very close to being centrosymmetric, it is found that it is often possible to obtain significant values of the Flack parameter. A theoretical basis for this surprising result is established. It has also been possible to establish an a priori estimate of the standard uncertainty of the Flack parameter based only on the chemical composition of the compound and the wavelength of the radiation. The paper concludes with preliminary presentations of bias in the Flack parameter and of inconsistent chemical and crystallographic data.

Density and structural changes in the bone of growing rats after weekly alendronate administration with and without a methotrexate challenge.

Alendronate (ALN) and other bisphosphonates have been used successfully in pediatric patients with osteopenia secondary to connective tissue diseases. Loss of growth in height has not been reported, but concerns remain regarding the effect of these potent antiresorptive agents when used in children and adolescents. High-dose methotrexate (MTX) and other chemotherapy drugs have been implicated in osteoporosis and a high fracture incidence in survivors of childhood cancers and are also associated with osteopenia in adult animals. The effect of high dose MTX on bone density during rapid skeletal growth, however, has not been widely studied, nor has the potentially therapeutic effect of bisphosphonates in this setting. We examined the effects of ALN and MTX administration, alone and in combination, on bone density, morphology, mechanical strength, and longitudinal growth in normal growing rats. Sprague-Dawley rats were given ALN once weekly (0.3 mg/kg) from 5 to 11 weeks of age, with and without a course of methotrexate (MTX) given daily in weeks 1 and 3 (0.75 mg/kg/day). Twenty-four animals were randomly divided into four groups: Control (vehicle), ALN alone, ALN + MTX, and MTX alone. After 6 weeks, the femora, tibiae, and lumbar spine were studied by dual-energy X-ray absorptiometry, peripheral quantitative computed tomography, mechanical strength testing, microradiography, light microscopy, and by determination of ash weights and bone lengths. ALN treatment increased bone mineral density (BMD) by 23% to 68%. The largest increases in the femur occurred in the distal third where endochondral bone growth was greatest and included large increases in trabecular bone and total cross-sectional area. ALN + MTX produced similar effects to ALN alone. MTX only reduced BMD by 8% in the vertebrae, but not significantly at other sites. MTX also led to femoral length reductions of 2.9%. The small reductions in BMD due to MTX were overwhelmed by the increases due to ALN, whereas the length loss was unaffected. Transverse density banding corresponding to weekly ALN administrations were clearly evident radiographically throughout the growing skeleton, likely due to decreased resorption and possibly increased mineralization in the bands. ALN or ALN + MTX treatment also led to increases in mechanical strength in the femora. Although MTX administration during growth leads to some BMD reduction, ALN given with MTX eliminates this reduction and in fact bone density and strength increase above control levels.

Tyrosine hydroxylase immunoreactivity in the developing visual pathway of the zebrafish.

We analyzed the distribution of tyrosine hydroxylase immunoreactivity in the central nervous zones involved in the processing of visual information during zebrafish ontogeny, employing a segmental approach. In the retina, we observed immunolabeled cells in the inner nuclear layer after hatching. From the juvenile stages onwards, some of these cells presented two immunolabeled processes towards the inner and outer plexiform layers of the retina, which are identified as interplexiform cells. In the adult zebrafish retina, we have identified two cellular types displaying immunoreactivity for tyrosine hydroxylase: interplexiform and amacrine cells. In the optic tectum, derived from the mesencephalon, no immunolabeled neurons were observed in any of the stages analyzed. The periventricular gray zone and the superficial white zone display immunostained neuropile from the end of fry life onwards. At the 30-day postfertilization, the tyrosine hydroxylase immunoreactive neuropile in the optic tectum presents two bands located within the retinorecipient strata and deeper strata, respectively. All diencephalic regions, which receive direct retinal inputs, show immunolabeled cells in the preoptic area, in the pretectum, and in the ventral thalamus from embryonic stages onwards. During the fry development, the immunolabeled neurons can be observed in the periventricular pretectum from 15-days postfertilization and in both the ventrolateral thalamic nucleus and suprachiasmatic nucleus from 30-days postfertilization. The transient expression of tyrosine hydroxylase is observed in fibers of the optic tract during fry and juvenile development. The existence of immunolabeled neuropile in the zebrafish retinorecipient strata could be related to the turnover of retinotectal projections.

Potential relationship between herpes viruses and rheumatoid arthritis: analysis with quantitative real time polymerase chain reaction.

OBJECTIVE: To determine whether the human herpes viruses, cytomegalovirus (CMV), Epstein-Barr virus (EBV), and human herpesvirus 6 (HHV-6), are detectable in serum and peripheral blood mononuclear cells (PBMCs) of patients with rheumatoid arthritis (RA). METHODS: 133 PBMC samples (61 RA, 72 healthy donors) and 136 serum samples (59 RA, 77 healthy donors) were analysed by quantitative real time polymerase chain reaction for DNA prevalence and viral load of HHV-6, EBV, and CMV. RESULTS: For PBMC samples significant differences were found for EBV in DNA prevalence (56% in RA v 33% in controls, p = 0.009) and viral load (copies/microg DNA 0-592.3 for RA v 0-40.4 for controls, p = 0.001). For serum samples a significant difference was found for HHV-6 DNA prevalence (10% in RA v 0% in controls, p = 0.006) and viral load (copies/microg DNA 0-529.1 for RA v 0 for controls, p = 0.007). CONCLUSIONS: Herpes viruses may have a role in RA, although alternative explanations are possible: (a) defects in cellular immunity in patients with RA may result in a relatively high viral load; (b) patients with RA may be more prone to infection/reactivation. The usefulness of monitoring the DNA viral load in patients with RA is questioned by these data.

Human parvovirus B19, varicella zoster virus, and human herpes virus 6 in temporal artery biopsy specimens of patients with giant cell arteritis: analysis with quantitative real time polymerase chain reaction.

OBJECTIVE: To evaluate the role of parvovirus B19 (B19), varicella zoster virus (VZV), and human herpes virus 6 (HHV-6) in the aetiopathology of giant cell arteritis (GCA). METHODS: Temporal artery biopsy specimens from 57 patients with GCA and 56 controls were investigated. DNA was obtained by biopsy, and quantitative real time polymerase chain reaction assay performed to establish the prevalence and viral load of B19, VZV, and HHV-6. Amplification of the human beta-globin gene was used as internal positive control. RESULTS: (a) B19 was detected in 31/57 (54%) patients (median viral load 45.2 (25th-75th centiles 0-180.2) copies/microg DNA) v 21/56 (38%) controls (median viral load 0 (0-66.7) copies/microg of DNA; p = 0.07 for DNA prevalence, p = 0.007 for viral load. Among 31 B19 positive samples, 21 (68%) patients with biopsy proven GCA had >10(2) B19 copies/microg of DNA v 5/21 (24%) controls; p = 0.001. (b) No significant difference was found for VZV (p = 0.94 for DNA prevalence; p = 0.76 for viral load) and HHV-6 (p = 0.89 for DNA prevalence; p = 0.64 for viral load) in the GCA group compared with controls. CONCLUSION: B19 may have a role in the aetiopathology of GCA, particularly in those patients with high viral load; no evidence was found for VZV and HHV-6.

Effects of axotomy on the expression of NADPH-diaphorase in the visual pathway of the tench.

The distribution of NADPH-diaphorase (ND) positive elements was analyzed throughout the visual pathway of the tench in normal conditions and after optic nerve transection. In the control retina, ND-labeled elements were observed in the photoreceptor, inner nuclear, outer nuclear and ganglion cell layers. In the optic nerve of control animals, small and numerous ND-positive glial cells that were identified as presumably astrocyte-like cells were observed. In the optic tracts and optic tectum, a different type of ND-positive glial cell was detected. Axotomy induced severe changes in the ND staining pattern in the visual pathway. A decrease in the number of ND-stained cells was detected in the retina. In the optic nerve of lesioned animals, the number of small cells gradually decreased, whereas the number of large cells did not change. Two new ND-positive cell populations were observed after the lesion: microglial-like cells appeared close to the lesioned area from 24 h to 7 days after transection, and astrocyte-like cells were found throughout the optic nerve from 14 days up to at least 120 days. The total number of ND-stained glial cells increased at 30 and 60 days and returned to control parameters at 120 days. In addition, the number of ND-positive cells increased at the same survival times in the optic tracts and in the retinorecipient strata of the optic tectum with respect to control animals. Thus, degenerative/regenerative processes in the fish visual pathway are accompanied by an increase in the number of ND-positive cells. Synthesis of nitric oxide is elicited in microglial-like cells as a response to axon injury, whereas the expression in astrocyte-like cells seems to be associated with both normal processes under physiological conditions and with the regenerative phase after the lesion.

Molecular structure and cytotoxicity of 3D-transition metal complexes capable to form a stable metal-nitrogen bond.

The cytotoxicity of several Co(II), Ni(II), Cu(II) and Zn(II) complexes with various molecular structures and geometries, has been tested on LoVo and 2008 cells at 1-100 microM concentration for 24 h exposure. On the basis of 24 h results, the exposure time was prolonged to 48 and to 72 hours. The most potent complexes result [Cu(tren)(H2O)]2+ 2Cl-, E, [CoCl3(H2Meppz)], G, and [CoCl3(HMe2ppz)], H, (tren=tris(2-aminoethyl)amine, H2Meppz=1-methylpiperazin-1-ium, HMe2ppz=1,4-dimethylpiperazin-1-ium cations). Nevertheless, these complexes are able to induce cell growth reduction of about 50% at highest doses tested (1-100 microM ) and after 72 h exposure.

[Single coronary artery with a right origin: angiography identification of the R-II subgroup]. / L'arteria coronaria singola di tipo destro: identificazione angiografica del sottogruppo R-II.

This case report illustrates the clinical and angiographic findings of 2 patients undergoing coronary angiography for ischemic heart disease and with the unexpected presence of anomalous origin of the left coronary artery from the right aortic sinus. The angiographic classification of the different subgroups of single coronary artery is reviewed and the 2 cases are identified as type R-II with septal and anterior course of the left main stem.

Experimental infection of Calomys callosus (Rodentia, Cricetidae) by Toxoplasma gondii.

Calomys callosus, Rengger 1830 (Rodentia, Cricetidae), a wild rodent found in Central Brazil, was studied to investigate its susceptibility to Toxoplasma gondii experimental infection and its humoral immune response against this protozoa. The electrophoretic profile of the serum proteins of C. callosus showed that IgG, which shows no affinity to Protein A, has higher cross reactivity with rat IgG than with IgG from other rodents. The susceptibility assay was performed by inoculation groups of animals with various suspensions of T. gondii tachyzoites from 10(2) to 10(6) parasites. All animals died between 3 and 9 days after infection and the kinetics of antibody synthesis was determined. Basically, they recognized predominantly the immunodominant antigen SAG-1 (P30). The immunohistochemistry assays revealed that the liver was the most heavily infected organ, followed by the spleen, lungs, intestine, brain and kidneys. It can be concluded that C. callosus is an excellent experimental model for acute phase of Toxoplasma infection.