Enhanced trafficking to the pancreatic lymph nodes and auto-antigen presentation capacity distinguishes peritoneal B lymphocytes in non-obese diabetic mice.

AIMS/HYPOTHESIS: NOD.Igmicro ( null ) mice lacking mature B cells are highly resistant to diabetes and display poor CD4 T cell responses to autoantigens. Nevertheless, the degree to which different B cell subsets contribute to diabetes in NOD mice remains unresolved. Due to their role in the recognition of microbial and autoantigens, peritoneal B cell characteristics were examined in NOD mice to see if they differ developmentally, phenotypically or functionally in aspects relevant to diabetogenesis. METHODS: The population dynamics, activation state, migratory behaviour and antigen presentation function were investigated in NOD peritoneal B cells. RESULTS: NOD peritoneal B cells were found to express abnormally high levels of co-stimulatory molecules (CD40, CD86 and CD69). In contrast, the expression of L-selectin and integrin alpha4beta1 was markedly reduced in NOD mice compared with BALB/c and C57BL/6 mice. The number of B cells in the peritoneum was lower in NOD than in control mice throughout development; migration of B cells from the peritoneum to the pancreatic lymph nodes in NOD mice was enhanced tenfold. NOD B cells showed no chemotactic response to sphingosine-1-phosphate, which normally acts to retain B cells in the peritoneum. Peritoneal B cells of NOD mice also presented insulin autoantigen to CD4 T cells, inducing T cell proliferation. CONCLUSIONS/INTERPRETATION: NOD peritoneal B cells are hyperactivated, migrate to the pancreatic lymph nodes and are capable of driving insulin-specific CD4 T cell activation. These characteristics could make them important for inducing or amplifying T cell responses against islet-antigens.

An extra cysteine in one of the non-calcium-binding epidermal growth factor-like motifs of the FBN1 polypeptide is connected to a novel variant of Marfan syndrome.

We present here a family with a clinical phenotype resembling Marfan syndrome (MFS), and displaying joint contracture and episodes of knee joint effusions, but lacking the cardiovascular features of the syndrome. The phenotype of this family represents a unique mixture of connective tissue symptoms, some of which are found in classical MFS and some of which are typical of dominant ectopia lentis. Linkage analyses suggested a linkage (LOD score 2.4; theta = 0) between the phenotype of the family and a polymorphic marker in the vicinity of the fibrillin locus on chromosome 15 (FBN1). Furthermore, a novel transition mutation was identified in the FBN1 gene in all the affected members of the family. In contrast to the majority of fibrillin mutations reported so far, this mutation substitutes a cysteine for arginine, producing an extra cysteine in one of the non-calcium-binding EGF-like motifs of the fibrillin polypeptide, most probably disturbing the formation of one of the three disulfide bridges known to be essential for the normal conformation of this motif.

Peptide immunoreactive neurons in the human retina.

The distribution of peptide-immunoreactive neurons in the human retina was investigated. Neurons displaying immunoreactivity towards substance P, vasoactive intestinal polypeptide (VIP), somatostatin, neuropeptide Y (NPY) and peptide histidine-isoleucine (PHI) were found in amacrine cells with cell bodies situated in the innermost part of the inner nuclear layer and nerve fibers ramifying in the inner plexiform layer in a manner differing according to the peptide investigated. Two other cell types were found. In the middle of the inner plexiform layer cell bodies showing immunoreactivity towards substance P, VIP and PHI were found. In the ganglion cell layer there were cell bodies showing immunoreactivity towards substance P, somatostatin, VIP and NPY. Substance P immunoreactive, somatostatin and NPY immunoreactive fibers situated at the border between the inner nuclear and outer plexiform layers and traversing the inner nuclear layer were also found.

Glucagon and VIP in the retina.

Immunoreactive glucagon and immunoreactive vasoactive intestinal polypeptide (VIP) have been demonstrated in neuronal elements in the retina of a number of species by immunohistochemistry. In the present study, the concentrations of glucagon-like and VIP-like material in retinae from different species were determined by radioimmunoassay. The retinal concentration of glucagon-like immunoreactivity was 10-35 pg/mg in goldfish, chicken, pigeon, and frog, whereas retinae from cow, pig, rabbit, and rat contained very little. Retinae from the latter four species were on the other hand rich in VIP-like material whereas retinae from cat, guinea pig, and goldfish contained very little. The glucagon-like immunoreactive material in chicken and frog retina was subjected to gel chromatography and high performance liquid chromatography (HPLC). The results indicate that the extracted protein is of similar molecular size as porcine pancreatic glucagon, distinct from porcine glicentin. VIP immunoreactive material extracted from bovine retina was similar in molecular size as authentic porcine VIP by gel chromatography.

Neuropeptide Y immunoreactive neurons in the guinea-pig uvea and retina.

Neuropeptide Y (NPY) is a recently discovered, amidated 36 amino acid residue neuropeptide present in many but not all sympathetic noradrenergic neurons. In the guinea-pig eye, NPY immunoreactive fibers were found to have the same distribution as noradrenergic fibers except that there were fewer at the iris dilator, in the cornea, and in the chamber angle. In the anterior uvea, the NPY immunoreactive fibers disappeared after excision of the homolateral superior cervical sympathetic ganglion, whereas in the choroid, many NPY immunoreactive fibers remained, indicating that they originate elsewhere. NPY immunoreactivity thus is not found in all sympathetic adrenergic neurons nor is it found only in such nerve fibers. In the retina, NPY immunoreactive fibers formed a single layer of processes in sublamina 1 of the inner plexiform layer. NPY immunoreactive cell bodies were found in the innermost cell row of the inner nuclear layer. The immunoreactivity was concentrated to the hillock region of these cells.

Simultaneous immunohistochemistry and autoradiography of peptides and 5-hydroxytryptamine in bird retina.

A method was developed for the simultaneous demonstration of two specific cellular constituents. Cryostat sections of formaldehyde fixed tissue from retinae treated with ((3)H)-5-hydroxytryptamine were subjected to immunohistochemistry and subsequent autoradiography. The method takes advantage of the very efficient and specific uptake mechanism that many types of neurons possess which makes it possible to label them with radioactivity. With this method a study was made on the possible co-occurrence in the avian retina of 5-hydroxytryptamine on one hand and somatostatin, glucagon and substance P on the other. Substance P and 5-hydroxytryptamine were investigated in pigeon retina whereas 5-hydroxytryptamine and somatostatin or glucagon were investigated in chicken retina. Though a large number of cell bodies were examined no co-occurrence of 5-hydroxytryptamine and any of the peptides was found. The sensitivity of the method allows an assertion that if present, double labelled neurons are likely to number less than 0.5-1% of the respective populations.

Immunohistochemical and quantitative analysis of 5-hydroxytryptamine in the retina of some vertebrates.

5-Hydroxytryptamine immunoreactive neurons were found in retinae from chicken, pigeon, frog and goldfish. They were localized among the amacrine cells with a distribution of cell bodies and nerve fibres that varied with the species. In chicken and pigeon, bipolar-like cell bodies were also found in the middle of the inner nuclear layer, sending processes inwards to the inner plexiform layer and outwards to the horizontal cells. The signalling direction of these cells is doubtful. No 5-hydroxytryptamine immunoreactivity was found in retinae from cow, pig, cat, rabbit, guinea-pig, rat or mouse. Quantitative analyses were performed with HPLC on extracts from chicken, pigeon, frog and goldfish retinae. High concentrations were found in goldfish and frog whereas less, about 100 ng/g, was observed in chicken and pigeon. The results suggest that 5-hydroxytryptamine is the transmitter of a set of amacrine cells in cold-blooded vertebrates and perhaps also in birds. The transmitter of the indoleamine accumulating neurons of mammals remains to be further elucidated.

Incidence of registered visual impairment in the Nordic child population.

A collaborative, population based, prospective register study on the incidence of visual impairment in children during the year 1993 was carried out in five Nordic countries with a total population of 17 million inhabitants. The child population was 3.8 million individuals aged 0-17 years. The following variables were taken into account: nationality, age, sex, diagnoses, aetiology, degree of visual impairment, and additional impairments. Classification routines from an earlier prevalence study were used. The present study included 304 children corresponding to an incidence of notification of 8/100,000 children, varying from 5.7 to 11.1 in the five countries. Fifty per cent of the visually impaired children were reported before they were 3 years of age. In approximately 45% of the children, visual impairment was due to various brain disorders, with cerebral amblyopia and secondary optic atrophy as the two leading causes. The relative impact of retinopathy of prematurity had decreased from the third most frequent cause (10%) in the prevalence study to seventh place (4%) in the incidence study. Two thirds of the children had additional impairments and these children also suffered from the most severe visual impairments. Among aetiological factors the majority (64%) were prenatal. The overall male:female ratio of 1.4:1 was identical to the sex ratio of the prevalence study.

Intrafamilial variation of the phenotype in Bardet-Biedl syndrome.

AIMS: To describe the variation of the phenotype within families with several individuals with Bardet-Biedl syndrome. METHODS: The phenotypes of affected siblings in 11 Scandinavian families with two or more members who had at least three of the features: retinal dystrophy, polydactyly, obesity, hypogenitalism, and mental retardation, were compared [corrected]. Individuals without retinal dystrophy were excluded. RESULTS: Intrafamilial variation of expressivity of the features obesity, polydactyly, abnormal radiograms of the extremities, hypogenitalism, short stature, paraplegia, and dental abnormalities was found. The retinal dystrophy varied with respect to both the onset of symptoms and the course of the disease. The morphology of the fundus, however, was consistent within the families. The disorder showed statistically significant genetic linkage to the BBS4 locus on chromosome 15 in the affected siblings in two of the families, but the clinical features in these patients did not differ from the other cases of Bardet-Biedl syndrome. CONCLUSION: Comparison of siblings with the Bardet-Biedl syndrome showed variation of the typical features. In addition, the course of retinal dystrophy varied. No distinctive clinical features were found to separate the BBS4 phenotype from the remaining patients.

Sigma-1 receptor agonist PRE084 is protective against mutant huntingtin-induced cell degeneration: involvement of calpastatin and the NF-κB pathway.

Alterations in mitochondria and increased oxidative stress are associated with the disease progression in Huntington's disease (HD). Endoplasmic reticulum (ER) stress and oxidative damage are linked through the close communication between the ER and mitochondria. Sigma-1 receptor (Sig-1R) is a chaperone protein in the ER that is involved in ER stress regulation, but little is known about its role in HD or the mechanisms for cell protection. Here we show that the Sig-1R agonist, PRE084 increases cell survival and counteracts the deleterious effects caused by N-terminal mutant huntingtin proteins in neuronal PC6.3 cells. Particularly, PRE084 increased the levels of cellular antioxidants by activating the NF-κB pathway that is compromised by the expression of mutant huntingtin proteins. These results show that the Sig-1R agonist has beneficial effects in models of HD and that compounds affecting the Sig-1R may be promising targets for future drug development in HD.

Visual impairment in Swedish children. III. Diagnoses.

PURPOSE: To gain an overview of the spectrum of diagnoses among Swedish visually impaired children. METHODS: An epidemiological study of all known visually impaired children was made by review of medical records. RESULTS AND CONCLUSION: In all we found 2373 children, 0-19 years of age, with an age-specific prevalence of 10.9/10,000. The two largest diagnostic groups included neuro-ophthalmological and retinal diseases. The most frequent disorders were cerebral visual impairment, non-hereditary optic atrophy, retinal dystrophy (when regarded as a general entity), congenital hypoplasia of the optic nerve and congenital cataract. Nystagmus secondary to brain disorder, albinism, congenital nystagmus, retinopathy of prematurity and high myopia were also found in a considerable number of patients. The leading diagnoses in children with WHO-defined childhood blindness were non-hereditary optic atrophy, cerebral visual impairment and retinopathy of prematurity. A large proportion of the children, especially in the groups with neuro-ophthalmological disorders and malformations of the posterior segment had additional impairments, emphasizing the importance of a multi-disciplinary approach when assessing multi-handicapped children.

Visual impairment in Swedish children. I. Register and prevalence data.

Knowledge of the epidemiology of visual impairment in children forms one of the cornerstones of pediatric ophthalmology. In contrast to e.g. the other Nordic countries, Sweden has had no efficient and continuous registration of visually impaired children. An epidemiological study on this subject has now been done, collecting data on all children and adolescents registered at the Low Vision Clinics throughout Sweden. In this work data on 2373 visually impaired children, 0-19 years of age and living throughout Sweden are presented. Data collected in the study include sex, date of birth, county, ocular diagnosis, systemic diagnosis, classification of visual impairment, aetiology and possible additional impairments. The prevalence of visual impairment as defined by WHO was 10.9/10,000 inhabitants in the current age group. A slight male preponderance was seen compared to the total population 0-19 years. This was not reduced when only non-genetic aetiological factors were taken into consideration. A total of 45% of the children had only a moderate visual impairment (WHO category 1), whereas approximately 25% were found in WHO categories 3,4 and 5, i.e. fulfilled the requirements for blindness. Additional impairments were found in 60% of the children. Mental impairment in combination with motor impairment or mental impairment exclusively were the most common ones seen.

Visual impairment in Swedish children. II. Etiological factors.

The analysis of etiological factors in a group of visually impaired children is of considerable importance when trying to find guidelines for possible preventive work. In this study we present etiological data on 2373 Swedish children. Data have been obtained by reviewing medical records on all known children with visual impairment throughout the country. In accordance with similar studies from industrialised countries, the group with prenatal etiology was the predominant, comprising 64% of the material. Within this group, half the patients had a disease of genetic origin. A total of 50% of all patients with prenatal etiology had an additional impairment, but in the group with diseases of genetic origin this proportion was smaller, only 40%. On the other hand, many children with additional impairments were found among those with an unspecified prenatal influence. Peri-/neonatal etiologies were found in 20% of the patients. In this group as many as 83% had additional impairments. This was even more pronounced among children delivered at term. The group with infantile/juvenile etiologies was small, 7%, with additional impairments in 66%. In 9% of all patients the etiology was classified as unknown. Among these, 80% had additional impairments. The visual impairment tended to be more pronounced, the later the disease was acquired. A male preponderance was seen in most etiological subgroups and in the material as a whole.

Electroretinograms in patients with achromatopsia.

Eleven patients with X-linked and 9 patients with autosomal recessive achromatopsia were examined with full-field electroretinograms. In the standard full-field ERG's, normal rod responses were obtained, but the amplitude of the cone b-waves was not detectable. With computer averaging and narrow bandpass filtering, residual cone b-wave responses could be detected in 10 of the 20 patients. The residual cone b-wave amplitudes were markedly different in the 3 families with X-linked achromatopsia. In two of them, residual cone b-wave responses were seen in all patients examined. In contrast, such responses were seen only in 2 of 7 patients in the third family. There were also differences in other clinical observations (mainly in the visual acuity and refractive error) and we therefore suggest that there are at least two forms of X-linked achromatopsia. The ratios of the cone response amplitudes to 30 Hz flickering orange and blue-green light suggested that the defect in the X-linked achromatopsia patients was of the protanope type, whereas in the autosomal patients, both the protanope and the deutanope type was seen. In conclusion, measurements of the residual cone b-wave amplitude responses are of diagnostic and may possibly be of prognostic value when examining children and other members of families with achromatopsia.

Glucagon immunoreactive neurons in the retina of different species.

Neurons displaying glucagon immunoreactivity were detected among the amacrine cells in the retina of goldfish, frog and pigeon. Nerve cell bodies were located in the inner nuclear layer with their processes ramifying in 2-3 more or less well-defined sublayers in the inner plexiform layer. The distribution of cell bodies and processes varied with the species. In pigeon retina two separate populations of glucagon immunoreactive neurons were found among the amacrine cells. In frog retina glucagon immunoreactivity was also discerned in cell bodies in the ganglion cell layer. These cell bodies sent processes outwards to the inner plexiform layer. No glucagon immunoreactive neurons were detected in the retina of the rat, rabbit, cat, pig or cow.