Gene conversion is a likely cause of mutation in PKD1

Approximately 70% of the gene responsible for the most common form of autosomal dominant polycystic kidney disease ( PKD1 ) is replicated in several highly homologous copies located more proximally on chromosome 16. We recently have described a novel technique for mutation detection in the duplicated region of PKD1 that circumvents the difficulties posed by these homologs. We have used this method to identify two patients with a nearly identical cluster of base pair substitutions in exon 23. Since pseudogenes are known to be reservoirs for mutation via gene conversion events for a number of other diseases, we decided to test whether these sequence differences in PKD1 could have arisen as a result of this mechanism. Using changes in restriction digest patterns, we were able to show that these sequence substitutions are also present in N23HA, a rodent-human somatic cell hybrid that contains only the PKD1 homologs. Moreover, these changes were also detected in total DNA from several affected and unaffected individuals that did not harbor this mutation in their PKD1 gene copy. This is the first example of gene conversion in PKD1 , and our findings highlight the importance of using gene-specific reagents in defining PKD1 mutations.      

Development of ocular dominance columns in the absence of retinal input

The initial establishment of ocular dominance columns in visual cortex is believed to involve the segregation of overlapping geniculocortical axons into eye-specific patches based on patterns of correlated activity. However, we found that total removal of retinal influence early in visual development did not prevent segregation of geniculocortical axons into alternating stripes with periodicity normal for ocular dominance columns. Because the patterning of geniculocortical afferents resists this dramatic change in the level, source and pattern of spontaneous activity, we propose that formation of ocular dominance columns relies on molecular cues present on thalamic axons, cortical cells or both.     

From dud to stud: copulatory behavior elicited through conditioned arousal in sexually inactive male rats

Sexually inactive male rats were conditioned to associate tones with electric shock to the flanks. In subsequent tests, when only tones were presented in the presence of an estrous female, these males began to copulate, pacing their behavior to the tone presentations. Only one animal tested for sexual behavior without the tones mated. It is suggested that the momentary augmentation of nonspecific arousal can lead to the initiation and subsequent pacing of mating responses in noncopulators.     

Biochemical mapping of the noradrenergic ventral bundle projection sites: Evidence for a noradrenergic-dopaminergic interaction

Norepinephrine (NE) and dopamine (DA) concentration and dopamine turnover were measured 12 days after a unilateral or bilateral noradrenergic ventral bundle (VB) transection to determine the noradrenergic projection sites and possible interactions with dopaminergic systems.Both bilateral and unilateral VB transection resulted in a significant reduction of NE of the nucleus accumbens, lateral septal nucleus, medial forebrain bundle, ventromedial nucleus, dorsomedial nucleus and medial amygdaloid nucleus. Bilateral transection also decreased NE content of the median eminence and the periventricular and arcuate nuclei. In the medial preoptic nucleus, the nucleus interstitialis striae terminalis and the central gray catecholamine area, bilateral transection significantly decreased NE concentrations while unilateral lesions had no significant effect. The anterior hypothalamic, lateral preoptic, and paraventricular nuclei responded to bilateral VB transection with a decrease in NE concentration and to unilateral lesion with a bilateral increase in NE. In the dorsal hippocampus and the caudate nucleus, bilateral lesions had no effect on NE concentrations while unilateral transection significantly decreased NE concentrations. Regions in which neither bilateral nor unilateral VB transection produced a significant change in NE content are the olfactory tubercle, the nucleus tractus diagonalis, substantia nigra pars compacta and reticulata, ventral tegmental area, habenula, superior colliculus, and the cingulate and piriform cortices.Transection of the noradrenergic ventral bundle also produced changes in dopaminergic systems suggesting a noradrenergic-dopaminergic interaction. Bilateral VB transection decreased the dopamine concentration and turnover in the nucleus accumbens, increased steady-state levels and turnover in the nucleus tractus diagonalis and increased dopamine concentration in the lateral septum. Unilateral VB transection decreased DA concentration bilaterally in the caudate nucleus, olfactory tubercle, nucleus accumbens and the nucleus interstitialis striae terminalis but increased concentrations in the substantia nigra pars reticulata (ipsilateral) and in the ventral tegmental area (bilateral). These results indicate a broad projection field for the noradrenergic ventral bundle and suggest a noradrenergic-dopaminergic interaction.     

Endovascular coil embolization of a spinal epidural arteriovenous fistula with associated cord compression from an enlarging venous varix

Spinal arteriovenous fistulas (AVFs) completely isolated to the epidural compartment are exceedingly rare. As such, the optimal management of these lesions is poorly defined. The aim of this technical note is to describe our endovascular technique for the occlusion of a purely epidural AVF of the thoracic spine associated with cord compression from an associated enlarging venous varix. A 40-year-old male presented with severe right-sided back pain and anterior thigh numbness after a sports-related back injury six months previously. Spinal magnetic resonance imaging (MRI) showed an enhancing, extradural mass lesion at T12. Spinal angiography revealed an epidural AVF supplied by a radicular branch of the right T12 subcostal artery and draining into the paravertebral lumbar veins, as well as an adjacent 20 × 13 mm² contrast-filling sac, compatible with a dilated venous varix. There was no evidence of intradural venous drainage. We elected to proceed with endovascular treatment of the lesion. At the time of embolization five days later, the venous varix had enlarged to 26 × 16 mm². The T12 epidural AVF was completely occluded with two coils, without residual or recurrent AVF on follow-up angiography one month later. The patient made a full recovery, and complete resolution of the venous varix and cord compression were noted on MRI at three months follow-up. Endovascular coil embolization can be successfully employed for the treatment of appropriately selected spinal epidural AVFs. Cord compression from an enlarging venous varix can be treated concurrently with endovascular occlusion of an associated spinal epidural AVF.     

Dendritic cells pulsed with protein antigens in vitro can prime antigen- specific, MHC-restricted T cells in situ [published erratum appears in J Exp Med 1990 Oct 1;172(4):1275]

T cells recognize peptides that are bound to MHC molecules on the surface of different types of antigen-presenting cells (APC). Antigen presentation most often is studied using T cells that have undergone priming in situ, or cell lines that have been chronically stimulated in vitro. The use of primed cells provides sufficient numbers of antigen-reactive lymphocytes for experimental study. A more complete understanding of immunogenicity, however, requires that one develop systems for studying the onset of a T cell response from unprimed lymphocytes, especially in situ. Here it is shown that mouse T cells can be reliably primed in situ using dendritic cells as APC. The dendritic cells were isolated from spleen, pulsed with protein antigens, and then administered to naive mice. Antigen-responsive T cells developed in the draining lymphoid tissue, and these T cells only recognized protein when presented on cells bearing the same MHC products as the original priming dendritic cells. In contrast, little or no priming was seen if antigen-pulsed spleen cells or peritoneal cells were injected. Since very small amounts of the foreign protein were visualized within endocytic vacuoles of antigen-pulsed dendritic cells, it is suggested that dendritic cells have a small but relevant vacuolar system for presenting antigens over a several day period in situ.     

Epigenetic modulation at birth – altered DNA-methylation in white blood cells after Caesarean section

Aim:  Delivery by C-section (CS) has been associated with increased risk for allergy, diabetes and leukaemia. Whereas the underlying cause is unknown, epigenetic change of the genome has been suggested as a candidate molecular mechanism for perinatal contributions to later disease risk. We hypothesized that mode of delivery affects epigenetic activity in newborn infants.
Methods:  A total of 37 newborn infants were included. Spontaneous vaginal delivery (VD) occurred in 21, and 16 infants were delivered by elective CS. Blood was sampled from the umbilical cord and 3–5 days after birth. DNA-methylation was analyzed in leucocytes.
Results:  Infants born by CS exhibited higher DNA-methylation in leucocytes compared with that of those born by VD (p < 0.001). After VD, newborn infants exhibited stable levels of DNA-methylation, as evidenced by comparing cord blood values with those 3–5 days after birth (p = 0.55). On postnatal days 3–5, DNA-methylation had decreased in the CS group (p = 0.01) and was no longer significantly different from that of VD (p = 0.10).
Conclusion:  DNA-methylation is higher in infants delivered by CS than in infants vaginally born. Although currently unknown how gene expression is affected, or whether epigenetic differences related to mode of delivery are long-lasting, our findings open a new area of clinical research with potentially important public health implications.     

From the horse's mouth: a case for Combined-Integrated doctoral training in professional psychology

Although new to many members of the psychological community, the notion of a "combined" model of training has been in existence for nearly 30 years (Beutler & Fisher, 1994). The present article is intended to outline the benefits and advantages of the Combined-Integrated (C-I) model. Following a brief discussion of the terms "combined" and "integrated," the advantages of C-I training are discussed across five key themes: (a) the high overlap across traditional practice area training, (b) the advantages of the breadth of training offered, (c) the advantages of C-I training for the professional of psychology, (d) the pragmatic advantages of C-I training to faculty and students, and (e) the benefits of C-I training for salient constituency groups.     

Predominance of null mutations in ataxia-telangiectasia

Ataxia-telangiectasia (A-T) is an autosomal recessive disorder involving cerebellar degeneration, immunodeficiency, chromosomal instability, radiosensitivity and cancer predisposition. The responsible gene, ATM, was recently identified by positional cloning and found to encode a putative 350 kDa protein with a Pl 3-kinase-like domain, presumably involved in mediating cell cycle arrest in response to radiation-induced DNA damage. The nature and location of A-T mutations should provide insight into the function of the ATM protein and the molecular basis of this pleiotropic disease. Of 44 A-T mutations identified by us to date, 39 (89%) are expected to inactivate the ATM protein by truncating it, by abolishing correct initiation or termination of translation, or by deleting large segments. Additional mutations are four smaller in-frame deletions and insertions, and one substitution of a highly conserved amino acid at the Pl 3-kinase domain. The emerging profile of mutations causing A-T is thus dominated by those expected to completely inactivate the ATM protein. ATM mutations with milder effects may result in phenotypes related, but not identical, to A-T.