Genetic disruption of the autism spectrum disorder risk gene PLAUR induces GABAA receptor subunit changes.

Disruption of the GABAergic system has been implicated in multiple developmental disorders, including epilepsy, autism spectrum disorder and schizophrenia. The human gene encoding uPAR (PLAUR) has been shown recently to be associated with the risk of autism. The uPAR(-/-) mouse exhibits a regionally-selective reduction in GABAergic interneurons in frontal and parietal regions of the cerebral cortex as well as in the CA1 and dentate gyrus subfields of the hippocampus. Behaviorally, these mice exhibit increased sensitivity to pharmacologically-induced seizures, heightened anxiety, and atypical social behavior. Here, we explore potential alterations in GABAergic circuitry that may occur in the context of altered interneuron development. Analysis of gene expression for 13 GABA(A) receptor subunits using quantitative real-time polymerase chain reaction (PCR) indicates seven subunit mRNAs (alpha(1), alpha(2), alpha(3), beta(2), beta(3), gamma(2S) and gamma(2L)) of interest. Semi-quantitative in situ hybridization analysis focusing on these subunit mRNAs reveals a complex pattern of potential gene regulatory adaptations. The levels of alpha(2) subunit mRNAs increase in frontal cortex, CA1 and CA3, while those of alpha3 decrease in frontal cortex and CA1. In contrast, alpha(1) subunit mRNAs are unaltered in any region examined. beta(2) subunit mRNAs are increased in frontal cortex whereas beta(3) subunit mRNAs are decreased in parietal cortex. Finally, gamma(2S) subunit mRNAs are increased in parietal cortex while gamma(2L) subunit mRNAs are increased in the dentate gyrus, potentially altering the gamma(2S):gamma(2L) ratio in these two regions. For all subunits, no changes were observed in forebrain regions where GABAergic interneuron numbers are normal. We propose that disrupted differentiation of GABAergic neurons specifically in frontal and parietal cortices leads to regionally-selective alterations in local circuitry and subsequent adaptive changes in receptor subunit composition. Future electrophysiological studies will be useful in determining how alterations in network activity in the cortex and hippocampus relate to the observed behavioral phenotype.

Disruption of Foxg1 expression by knock-in of cre recombinase: effects on the development of the mouse telencephalon.

The cre/loxP system is used routinely to manipulate gene expression in the mouse nervous system. In order to delete genes specifically from the telencephalon, the Foxg1-cre line was created previously by replacing the intron-less Foxg1 coding region with cre, resulting in a Foxg1 heterozygous mouse. As the telencephalon of heterozygous Foxg1 mice was reported to be normal, this genotype often has been used as the control in subsequent analyses. Here we describe substantial disruption of forebrain development of heterozygous mice in the Foxg1-cre line, maintained on the C57BL/6J background. High resolution magnetic resonance microscopy reveals a significant reduction in the volume of the neocortex, hippocampus and striatum. The alteration in the neocortex results, in part, from a decrease in its tangential dimension, although gross patterning of the cortical sheet appears normal. This decrease is observed in three different Foxg1 heterozygous mouse lines, independent of the method of achieving deletion of the Foxg1 gene. Although Foxg1 is not expressed in the diencephalon, three-dimensional magnetic resonance microscopy revealed that thalamic volume in the adult is reduced. In contrast, at postnatal day 4, thalamic volume is normal, suggesting that interactions between cortex and dorsal thalamus postnatally produce the final adult thalamic phenotype. In the Foxg1-cre line maintained on the C57BL/6J background, the radial domain of the cerebral cortex also is disrupted substantially, particularly in supragranular layers. However, neither Foxg1 heterozygous mice of the Foxg1-tet (tetracycline transactivator) line, nor those of the Foxg1-lacZ and Foxg1-cre lines maintained on a mixed background, displayed a reduced cortical thickness. Thus Cre recombinase contributes to the radial phenotype, although only in the context of the congenic C57BL/6J background. These observations highlight an important role for Foxg1 in cortical development, reveal noteworthy complexity in the invocation of specific mechanisms underlying phenotypes expressed following genetic manipulations and stress the importance of including appropriate controls of all genotypes.

Pathogen inactivation of Leishmania donovani infantum in plasma and platelet concentrates using riboflavin and ultraviolet light.

BACKGROUND AND OBJECTIVES: Leishmania is transmitted by the bite of the phlebotomine sandfly or by transfusion of infected blood products. Leishmaniasis currently poses a significant problem in several parts of the world, and is an emerging problem in others. The Mirasol PRT technology is based on the use of riboflavin and ultraviolet light to generate chemical reactions in the nucleic acids of pathogens, which prevents replication and leads to inactivation. The intent of this study was to examine the ability of the Mirasol PRT System to kill the Leishmania parasite in human plasma and platelet concentrates. MATERIALS AND METHODS: In visceral Leishmaniasis, amastigotes are present in the blood and in the reticuloendothelial system within monocytes. For each unit of plasma or platelets treated, isolated mononuclear cells obtained from 100 ml of normal donor whole blood were incubated with 1.0 x 10(8) Leishmania donovani infantum promastigotes to produce amastigote-laden macrophages. The infected macrophages were added to 250 ml of human plasma or to 250 ml of platelet concentrates. Infected units were cultured pretreatment in 10-fold serial dilutions to determine the limits of detection. Thirty millilitres of 500 microM riboflavin was added to each unit, which was then illuminated with 5.9 J/cm2 of ultraviolet light (6.24 J/ml). After treatment and after 2 months of frozen storage, plasma units were cultured in 10-fold serial dilutions. Platelets were cultured on the day of treatment and on day 5 of storage post-illumination. RESULTS: A 5 log reduction of Leishmania was demonstrated in five of six units of plasma, and a 7 log reduction of Leishmania was demonstrated in one plasma unit. A 5 log reduction of Leishmania was demonstrated in five of six units of platelets, and a 6 log reduction of Leishmania was demonstrated in one unit. CONCLUSIONS: There is no donor screen for Leishmania and other pathogens constantly emerging in our blood supply. The Mirasol PRT System for Platelets and Plasma is an effective means of killing Leishmania and other emerging pathogens in these blood products.

The vortex concentrator for suspended solids treatment.

The use of vortex concentrators is becoming increasingly popular for suspended solids reduction in combined sewer overflows and stormwater. This study is a laboratory investigation of the use of vortex concentrators to reduce the solids concentration of synthesized stormwater. The synthesized stormwater was made with water and addition of particles; sand, granular activated carbon, and sewer sediments. The vortex concentrator was made of acryl resin 300 mm in diameter. To determine the efficiency for various influent suspended solids (SS) concentrations, tests were performed with different SS concentrations. The samples were taken simultaneously at the influent storage tank and effluent tank, and measured SS concentrations. The range of surface loading rates were 120 to 850 m3/m2/day, and influent SS concentrations were varied from 300 to 5,000 mg/L. To determine the optimum coagulant dosage, jar tests were conducted with coagulants such as PAM and PAC. It was found that optimum coagulant and its dosage were PAM and 2 mg/L. The overall SS removal efficiency of the vortex concentrator for typical stormwater was estimated at about 65%. With an increase of SS concentration, the removal efficiency was increased. Since the SS concentration of stormwater was higher than 1,000 mg/L, the removal efficiency of the vortex concentrator for stormwater could be estimated to be 65-70%. The SS removal efficiency was increased with an increase of retention time, and the optimum retention time was 0.15-1.0 minutes. With an increase of the foul to overflow Q(F)/Q(o), a key parameter for vortex concentrator operation, the removal efficiency was increased. An alternative solution to improve treatment efficiency might be to set a follow-up retention basin. Based on a series of settling tests on the treated overflow water from the vortex concentrator, 5 to 10 minutes hydraulic retention time in a follow-up retention basin would substantially improve the results.

First flush analysis of urban storm runoff.

Stormwater runoff was monitored on 13 separate urban watersheds, which were chosen to represent distinct types of residential and industrial development, along with various watershed characteristics. A total of 38 storm events were monitored to investigate the first flush phenomenon. The first flush phenomenon may be defined as the initial period of stormwater runoff during which the concentration of pollutants is substantially higher than during later stages. The magnitude of the first flush phenomenon, and if it actually occurs, was calculated using a method of data analysis which results in determining the 'event mean concentration' (EMC). The magnitude of the first flush phenomenon was found to be greater for some pollutants (e.g. suspended solids from residential areas) and less for others (e.g. chemical oxygen demand from industrial areas). No correlation was observed between the first flush phenomenon and the antecedent dry weather period, however, the first flush phenomenon was greater for smaller watershed areas.

The injection of nodules of Dupuytren's disease with triamcinolone acetonide.

Over a 4-year period 63 patients (75 hands) with Dupuytren's nodules were treated with a series of injections with the steroid triamcinolone acetonide directly into the area of disease. The purpose of this study was to determine whether intralesional injections of triamcinolone acetonide could produce softening and flattening in nodules of Dupuytren's disease as seen in the intralesional injections of hypertrophic scars and keloids. After an average of 3.2 injections per nodule 97% of the hands showed regression of disease as exhibited by a softening or flattening of the nodule(s). Although some patients had complete resolution of the nodules, most experienced definite but incomplete resolution of the nodules in the range of 60% to 80%. Although a few patients did not experience recurrence or reactivation of the disease in the injected nodules or development of new nodules, 50% of patients did experience reactivation of disease in the nodules 1 to 3 years after the last injection, necessitating 1 or more injections. The findings of this study indicate that the intralesional injection of nodules of Dupuytren's disease with triamcinolone acetonide may modify the progression of the disease.

Twenty-five-year follow-up evaluation of an active silicone/Dacron tendon interposition prosthesis: A case report.

A 19-year-old man sustained a severe avulsion wound of the dominant distal forearm, dividing the radial and ulnar arteries, median and ulnar nerves, and all flexor tendons. Initial treatment consisted of revascularization. Shortly thereafter he had sural nerve grafting of the median and ulnar nerves. This was followed by insertion of a silicone/Dacron tendon interposition prosthesis to reconstruct a 4-cm deficit in the flexor profundus tendons and the flexor pollicis longus tendon. Six weeks thereafter an opposition transfer using the extensor indicis proprius and a Brand type 2 intrinsic transfer using the extensor carpi radialis longus and a plantaris tendon graft were performed. Several months later an attempt was made to remove the prosthesis. It was encased in scar tissue, however, and left in place. Evaluation 25 years later revealed that the flexor tendons and prosthesis were functioning well.

Autoimmune disease and chronic lymphocytic leukemia: autoimmune hemolytic anemia, pure red cell aplasia, and autoimmune thrombocytopenia.

Immune dysregulation, a hallmark of chronic lymphocytic leukemia (CLL), manifests itself in three autoimmune diseases: warm autoimmune hemolytic anemia (AIHA); idiopathic thrombocytopenia (ITP); and, pure red cell aplasia (PRCA). AIHA occurs in 11% of advanced stage CLL patients. Prednisone is the first treatment of choice, with 90% responses and 65% complete responses. More than 60% of patients relapse when treatment is stopped. Intravenous immunoglobulin, the next line of treatment, causes responses in 40% of patients. While the data are very limited, cyclosporine A is a reasonable choice for third-line therapy. Alkylating agents, danazol, plasma exchange, immunoabsorption, vincristine-loaded platelets, splenectomy, and splenic irradiation are also reported to cause responses. The data on mechanisms of AIHA are most consistent with immune dysregulation leading to loss of tolerance to a self antigen which in turn leads to the immune-based hemolytic anemia. PRCA is underrecognized in CLL with 6% of CLL patients having PRCA when tested for it. Unlike AIHA, PRCA often occurs in early stage disease. Anemia, reticulocytopenia, and a marrow virtually devoid of red blood cell precursors are hallmarks of PRCA. Corticosteroid therapy is the first line of treatment. If a response is not obtained in 4 weeks, cyclosporine A should be added. Although the data on pathophysiology are very limited, PRCA appears to be the result of an abnormal T cell that both fails in its normal function to support growth and inhibits the growth of erythroid progenitor cells. ITP occurs in 2-3% of CLL patients, occurs in early stage disease and may be a presenting manifestation. Initial therapy for ITP mirrors the guidelines for primary ITP. Initial therapy should consist of prednisone. Seventy percent of patients respond. Splenectomy is a reasonable second-line treatment. Autoimmune phenomena, largely related to blood cells, are based in the immune dysregulation of CLL. Longer survivals in CLL patients, more treatment regimens per patient, and more immunosuppression with modern treatments, allow us to predict an increasing incidence of autoimmune blood cell diseases in CLL.

The use of the full thickness skin graft in Dupuytren's contracture.

For those hand surgeons who have experienced early complications associated with limited fasciectomies (those who have not, have not done enough limited fasciectomies) and are frustrated by a high rate of recurrence or extension of the disease, incision of the cord and interposition of a full thickness graft is a technique to seriously consider. The procedure is not difficult to perform, but patience and attention to detail are prerequisites for success. That success is measured by complete or near complete release of the contracture with a minimum of morbidity, a nil recurrence rate, and extension rate of less than 10%. This technique is indicated for patients who have one or more elements of the Dupuytren's diathesis. Usually people older than 65 who develop Dupuytren's contracture do not have the diathesis, and their disease can be managed by limited fasciectomy and Z plasty skin lengthening. Finally, Dupuytren's disease presenting with no contracture can be managed effectively and conservatively by a series of intralesional injections of triamcinolone into the nodules and cords, the treatment of choice for all plantar nodules and knuckle pads.

Dermofasciectomy and full-thickness grafts in the treatment of Dupuytren's contracture.

We report a 16-year experience with full-thickness grafts to resurface defects created by releasing contractures of Dupuytren's disease. This technique was used in 68 patients with Dupuytren's contracture from 1970 to 1985. Follow-up of 36 hands of 24 patients averaged 3.9 years postoperatively. There was no recurrent disease in the palms and digits that were covered with the full-thickness grafts. The incidence of extension outside the grafts was 8%. The area of full-thickness grafting covered most of the width of the palm, an extension of Gonzalez's technique, which was presented in 1970.