The CC2D1A, a member of a new gene family with C2 domains, is involved in autosomal recessive non-syndromic mental retardation.

BACKGROUND: The molecular basis of autosomal recessive non-syndromic mental retardation (NSMR) is poorly understood, mostly owing to heterogeneity and absence of clinical criteria for grouping families for linkage analysis. Only two autosomal genes, the PRSS12 gene on chromosome 4q26 and the CRBN on chromosome 3p26, have been shown to cause autosomal recessive NSMR, each gene in only one family. OBJECTIVE: To identify the gene causing autosomal recessive NSMR on chromosome 19p13.12. RESULTS: The candidate region established by homozygosity mapping was narrowed down from 2.4 Mb to 0.9 Mb on chromosome 19p13.12. A protein truncating mutation was identified in the gene CC2D1A in nine consanguineous families with severe autosomal recessive NSMR. The absence of the wild type protein in the lymphoblastoid cells of the patients was confirmed. CC2D1A is a member of a previously uncharacterised gene family that carries two conserved motifs, a C2 domain and a DM14 domain. The C2 domain is found in proteins which function in calcium dependent phospholipid binding; the DM14 domain is unique to the CC2D1A protein family and its role is unknown. CC2D1A is a putative signal transducer participating in positive regulation of I-kappaB kinase/NFkappaB cascade. Expression of CC2D1A mRNA was shown in the embryonic ventricular zone and developing cortical plate in staged mouse embryos, persisting into adulthood, with highest expression in the cerebral cortex and hippocampus. CONCLUSIONS: A previously unknown signal transduction pathway is important in human cognitive development.

Increased mitotic activity in the dentate gyrus of the hippocampus of adult C57BL/6J mice exposed to the flurothyl kindling model of epileptogenesis.

Repeated flurothyl-induced generalized forebrain seizures result in a progressive and permanent lowering of the generalized seizure threshold in mice and an increase in the percentage of animals expressing forebrain-brainstem seizures, when rechallenged with flurothyl, after a stimulation-free period. Since this seizure paradigm serves as an excellent model for examining changes in seizure threshold and seizure propagation, we were interested in examining mitotic activity in hippocampal progenitors following flurothyl-induced epileptogenesis. In the present studies, we investigated (1). the effect of one or eight flurothyl-induced seizures on mitotic activity in the hippocampal dentate gyrus of adult mice measured by 5-bromo-2'-deoxyuridine incorporation, (2). the time course of change in hippocampal mitotic activity, (3). the cellular phenotype of these mitotically active cells, and (4). the relationship of changes in mitotic activity to changes in seizure threshold and phenotype. Significant increases in hippocampal mitotic activity were observed in mice exposed to either one or eight flurothyl-induced seizures. Increases were observed at 1 and 3 days following one seizure, and at 0, 1, 3, and 7 days following eight seizures. Confocal analyses, using neuronal and glial markers, suggest that the majority of these mitotic cells are neurons. In addition, no correlation was observed between hippocampal mitotic activity and the final seizure type that animals expressed following incubation and flurothyl retest. A significant correlation was observed between hippocampal mitotic activity and seizure threshold in flurothyl-kindled mice. Overall, these results indicate that both one and eight flurothyl-induced seizures are potent inducers of hippocampal neurogenesis in adult mice. Results further suggest that the increases in hippocampal neurogenesis are not directly related to the processes that underlie the shift in behavioral seizure phenotype, but may be involved in either the establishment or the maintenance of seizure threshold in this flurothyl model of epileptogenesis.

Patient satisfaction with laparoscopic Burch retropubic urethropexy.

OBJECTIVE: To assess six-month and three- to four-year patient-oriented outcomes after laparoscopic Burch retropubic urethropexy. STUDY DESIGN: Twenty-two women with urodynamically proven genuine stress incontinence with urethral hypermobility underwent laparoscopic Burch retropubic urethropexy. Preoperatively, all 22 women completed a questionnaire concerning their incontinence. Postoperative measures of symptoms of incontinence, impact of incontinence on daily activities and patient satisfaction were assessed at six months and three to four years postoperatively. In those women who were followed, the questionnaire data at the three-time points (preoperative, six months and three to four years) were compared. RESULTS: Thirteen women (59%) completed postoperative questionnaires at six months and three to four years. When compared to preoperative data, there was a significant improvement in symptoms of stress incontinence at six months (P = .0005) and at three to four years (P = .002). There was also a significant reduction in limitations on daily activities at six months (P = .0005) and at three to four years (P = .0005) as compared to preoperative data. Twelve of the 13 women considered their surgery successful at six months and at three to four years. CONCLUSION: After laparoscopic Burch retropubic urethropexy, there was a significant improvement in patient-oriented outcomes, including complaints of incontinence and functional status.

Bidirectional transfer between electrical and flurothyl kindling in mice: evidence for common processes in epileptogenesis.

PURPOSE: This study sought to determine whether there was a transfer of seizure susceptibility between two models of epileptogenesis, electrical kindling and a newly described model of flurothyl kindling. In this study, we determined the effects of preexposure to one kindling agent on the seizure responsiveness to the other. METHODS: Mice were divided into three groups: (a) six mice (FLK) were kindled with flurothyl, rechallenged with flurothyl after a 28-day incubation phase, implanted with olfactory bulb (OB) electrodes, and electrically kindled; (b) six mice (ELK) were implanted with OB electrodes, electrically kindled to six stage 5 seizures, and given one flurothyl trial 3 days later and a second flurothyl trial after a 28-day incubation period; and (c) six mice (IMP) were implanted with OB electrodes, tested with flurothyl at the same times as the ELK group, and later electrically kindled. RESULTS: Mice that were previously kindled with flurothyl (FLK) had significantly faster electrical kindling rates to one stage 5 seizure or to six stage 5 seizures compared with animals in the ELK and IMP groups. Mice that were previously exposed to either electrical kindling or flurothyl kindling had significantly diminished latencies to generalized seizure onset (flurothyl-induced seizure thresholds) either before or after a 28-day incubation period compared with the IMP control mice. In addition, both the FLK and ELK groups had significantly increased percentages of mice expressing forebrain-brainstem seizures, compared with the IMP group, following either rechallenge with flurothyl after a 28-day incubation or focal electrical kindling. CONCLUSIONS: These findings indicate a near-complete bidirectional transfer between these electrical and flurothyl kindling models. Mice that were previously exposed to either electrical or flurothyl kindling have increased seizure susceptibilities and altered seizure phenotypes when exposed to the other seizure paradigm. Overall, these studies indicate that previous seizures are the critical determinant of the bidirectional transfer of seizure susceptibility observed, and not the electrical or pharmacologic properties of the original kindling agent. Finally, the observation of near identity in transfer characteristics between electrical and flurothyl kindling models suggests that the proepileptogenic processes initiated by exposure to either model are similar.

The role of the ventromedial nucleus of the hypothalamus in epileptogenesis.

Flurothyl kindling initiates a time-dependent process that results in a facilitated propagation from the forebrain to the brainstem seizure system and in an increase in the complexity of behavioral seizure expression. We investigated the involvement of the ventromedial nucleus of the hypothalamus (VMH) in mediating this facilitated propagation between these seizure systems. Bilateral ibotenic acid lesions of the VMH, but not the dorsomedial nucleus of the hypothalamus (DMH), resulted in a disruption in the propagation of seizure activity from the forebrain to the brainstem. Moreover, VMH lesioned mice were able to express brainstem seizures following minimal corneal electroconvulsive shock (mECS). Together, our results indicate that the VMH is a critical substrate involved in propagating seizure activity between the forebrain and brainstem, but is not involved in the expression systems necessary for forebrain or brainstem seizure manifestations.

A role for the bilateral involvement of perirhinal cortex in generalized kindled seizure expression.

The perirhinal cortex (PRh) has been suggested as a substrate for the expression of generalized clonic seizures in the late stages of kindling development (stages 4-5). Using the induction of Fos as a marker of neuronal activation, the PRh region was investigated after kindling or nonkindling electrical stimulation. Nonkindling electrical stimulation of the PRh elicited stimulus-locked behaviors, without afterdischarge. These behaviors were characterized by rearing and bilateral forelimb clonus which were terminated upon electrical stimulus offset in half of the rats displaying this behavior (with the other half expressing self-sustained seizures). In these animals, Fos immunoreactivity was found throughout neocortical and subcortical structures in the hemisphere ipsilateral to the stimulating electrode. By contrast, Fos-immunoreactivity in the contralateral hemisphere was localized primarily in the PRh and frontal motor cortex. Likewise, similar patterns of Fos immunoreactivity were observed in both hemispheres of rats following kindling to one generalized clonic seizure from several limbic and paleocortical structures. These results suggest that the bilateral involvement of the PRh is critical in producing the bilateral behaviors associated with generalized clonic seizure expression. In support of this interpretation, infusion of 3 M KCl directly into the contralateral PRh of rats kindled to a single stage 4-5 (generalized clonic) seizure from the ipsilateral amygdala reduced seizure manifestations from a generalized clonic seizure (stage 4-5) to a unilateral clonic seizure (stage 3) without affecting measures of focal excitability. Taken together, these data indicate a role for the bilateral involvement of the PRh in generalized clonic seizure expression whether evoked from the naive or kindled state. These results further indicate that bilateral behaviors require the bilateral involvement of the structures necessary for the expression of these behaviors.

Decreased brainstem seizure thresholds and facilitated seizure propagation in mice exposed to repeated flurothyl-induced generalized forebrain seizures.

We recently have described a novel model of epileptogenesis utilizing the inhalant chemoconvulsant, flurothyl (Applegate et al., 1997; Samoriski and Applegate, 1997). The hallmark feature of this model is a change in behavioral seizure phenotype from a forebrain seizure, observed during the initial flurothyl exposures, to a brainstem seizure, elicited by flurothyl, after a 28-day stimulation free incubation period. In this study, we sought to establish the basis for this change in behavioral seizure response. To this end, we examined the effects of exposure to this paradigm on the generalized brainstem seizure threshold and on the propagation of forebrain seizures onto the brainstem seizure substrate. Ten mice were given flurothyl-induced generalized forebrain seizures on 8 consecutive days (induction phase). The other ten mice were not exposed to the flurothyl induction paradigm and served as controls. Minimal corneal electroconvulsive shock (mECS--20 mA) was used to assay whether there was any change in the animals' generalized brainstem seizure thresholds at 3, 14 and 28 days following the last flurothyl seizure trial. Mice that were exposed to flurothyl exhibited a progressive increase in the percentage of animals having a mECS-induced brainstem seizure when tested at 3 (40%), 14 (70%) and 28 (90%) days following the last flurothyl seizure. Control mice rarely had a brainstem seizure at any of the three time points tested, mostly forebrain seizures were observed. These results suggest that there is a significant progressive lowering of the brainstem seizure threshold, during the incubation phase of the flurothyl paradigm, which is coincident with the previously reported time course of change in the behavioral seizure phenotype observed using this flurothyl model (Applegate et al., 1997; Samoriski and Applegate, 1997). Following mECS testing, mice were implanted with bipolar electrodes and kindled from the olfactory bulb (OB). Mice exposed to the flurothyl paradigm demonstrated significantly faster kindling rates, longer afterdischarge durations. and longer durations of and latencies to stage 5 seizures compared to controls. Furthermore, animals exposed to the flurothyl protocol demonstrated an increase in the expression of brainstem seizures after focally-elicited OB afterdischarges. These results suggest that there is an increased interaction between the forebrain and brainstem seizure systems after exposure to this model of epileptogenesis. Together, results indicate that the change in behavioral seizure phenotype observed following exposure to our flurothyl paradigm are promoted by both decreases in brainstem seizure thresholds and facilitated forebrain seizure propagation onto the brainstem seizure system.

Analysis of the importance of controllable versus uncontrollable stress on subsequent behavioral and physiological functioning.

Original observations of the effects of stress exposure on behavioral, physiological and pathological indices were documented in the mid 1960s [J.B. Overmier, Interference with avoidance behavior: failure to avoid traumatic shock, J. Exp. Psychol. 78 (1968) 340-343 [12]; J.B. Overmier, M.E.P. Seligman, Effects of inescapable shock upon subsequent escape and avoidance learning, J. Comp. Physiol. Psychol. 63 (1967) 28-33 [13]; M.E.P. Seligman, S.F. Maier, Failure to escape traumatic shock, J. Exp. Psychol. 74 (1967) 1-9 [15]; J.M. Weiss, Effects of coping responses on stress, J. Comp. Physiol. Psychol. 65 (1968) 251-260 [18]]. Studies employing the triadic design (e.g. escapable stress, yoked-inescapable stress and no stress) indicated that the deficits following stress exposure were not caused by stress per se, rather the uncontrollability of the stress was the critical determinant. In this paradigm, the first group (escape) receives exposure to an environmental event that it can "control" by performing a behavioral response. Stress control or coping behavior includes the ability to alter the onset, duration, intensity or pattern of an aversive experience [S.F. Maier, M.E.P. Seligman, Learned helplessness: theory and evidence, J. Exp. Psychol.: Gen. 105 (1976) 3-46 [10]]. The second group is "yoked" to its escape partner and receives the identical physical stressor as its escape counterpart, but there is no behavioral response that the yoked subject can make to alter the outcome. The third group (naive) receives no stress exposure and is either restrained in the experimental apparatus or remains in the home cage until subsequent testing. Researchers using this triadic design should be aware of the concerns of certain investigators [R.M. Church, Systematic effect of random error in the yoked control design, Psychol. Bull. 62 (1964) 122-131 [3]; E.A. Wasserman, Response bias in the yoked control procedure, Behav. Brain Sci. 11 (1988) 477-478 [17]] who have raised important issues about the validity of the yoked control design because of the possibility of systematic biases. For example, individual differences in stress reactivity may result in random error in the yoked control group. This point will be addressed further in Section 5. This procedure allows the investigator to analyze the contributions of the importance of psychological dynamics of stress on a variety of dependent measures including: behavioral, pharmacological, neurochemical and immunological indices.

Laparoscopic-assisted vaginal hysterectomy using the laparosonic coagulating shears.

The use of the Laparosonic Coagulating Shears (LCS) for laparoscopic-assisted vaginal hysterectomy (LAVH) was evaluated in three women. The indications for surgery included chronic pelvic pain, adhesions, endometriosis, symptomatic uterine fibroids, and abnormal bleeding. The entire laparoscopic portion of the LAVH was performed with the LCS. All three patients had an uneventful postoperative course, and continue to do well one year postoperatively. The harmonic scalpel produces surgical incisions with concomitant hemostasis. With the introduction of the LCS, larger vessels can be managed safely. The LCS produces less charring and less thermal tissue injury, reduces postoperative adhesions, and promotes faster healing. The cavitational effect facilitates dissection, and the minimal heat production and absence of current through the patient contribute to safety. Ultrasonic activated technology is easy to use, cost effective, and affords the surgeon a greater margin of safety. This is only a preliminary report, and further study is needed, but the benefits of ultrasonically activated technology and the LCS are readily apparent.

Environmentally induced changes in peripheral benzodiazepine receptors are stressor and tissue specific.

The stress-induced changes in peripheral benzodiazepine receptors (PBR) can be observed in a number of different tissues, depending upon the nature and chronicity of the aversive experience. In addition, virtually all stress procedures that cause rapid changes in PBR simultaneously increase the physical activity or metabolic rate of the subjects. The present study analyzed the contributions of rapid alterations in activity or metabolic rate with and without aversive stimulation and their subsequent impact on PBR. Mechanically induced increases in activity by forced running stress results in a significant reduction in [3H]Ro 5-4864 binding to PBR in olfactory bulb, opposite to the PBR changes in this tissue following forced cold-water swim stress. Pharmacological induction of increased locomotor activity as well as metabolic rate by d-amphetamine causes a significant increase in cardiac PBR binding, again, opposite to the response typically observed following inescapable shock stress. Finally, administration of the anxiogenic beta-carboline, FG-7142, causes increases in both hippocampus and adrenal gland PBR binding reminiscent of acute noise stress exposure. These experiments demonstrate that increased locomotor activity or metabolic rate alone is not a necessary and sufficient condition for previous stress-induced changes in PBR. Conversely, increased metabolic rate coupled with an aversive stimulus appears to be an important factor for inducing stress-like changes in PBR. This data, coupled with previous reports, suggests that rapid alterations in these sites are stressor and tissue dependent. Finally, we propose that the PBR may be involved in many aspects of the stress response including: a) a blowarning system in adrenal gland, b) participation in stress-induced hypertension via renal PBR, and c) a modulator of stress-induced immunosuppression and subsequent recovery of function or recuperation by actions on immune cells.

The protective effects of stress control may be mediated by increased brain levels of benzodiazepine receptor agonists.

Control over stress protects against many of the deleterious effects of stress exposure, but the endogenous mediators responsible for these prophylactic effects have remained elusive. Using behavioral pharmacology, in vitro radioligand binding and neurochemical analyses, we demonstrate that exposure to escapable stress results in brain and behavior changes reminiscent of benzodiazepine administration. The stress control group shows significant protection against picrotoxinin-induced seizures, reductions in [35S]t-butylbicyclophosphorothionate (TBPS) binding and a 3-fold increase of benzodiazepine-like substances in brain in comparison to both yoked-inescapable shock and non-shock controls. These observations suggest that coping behavior leads to the release of endogenous benzodiazepine-like compounds in brain which protect the organism from stress pathology.

An ectopic pregnancy in the upper retroperitoneum following in vitro fertilization and embryo transfer.

Thirty-seven days after successful embryo transfer, a chorionic vesicle was recovered from the upper abdominal retroperitoneal space in a patient presenting with an acute life-threatening hemorrhage. This is the first case report of a retroperitoneal ectopic pregnancy which, although uncommon, is a potentially fatal complication of in vitro fertilization-embryo transfer.