Palladium-mediated 11C-carbonylations using aryl halides and cyanamide.

A robust and high-yielding radiochemical synthesis of 11C-N-cyanobenzamides using a palladium-mediated aminocarbonylation with 11C-CO, aryl halides and cyanamide is described. The bidentate ligand 1,1'-bis(diphenylphosphino)ferrocene provided 11C-N-cyanobenzamides from aryl-iodides, bromides, triflates and even chlorides in 28-79% radiochemical yield after semi-preparative HPLC. To further highlight the utility of this method, novel 11C-N-cyanobenzamide analogs of flufenamic acid, meflanamic acid, dazoxiben and tamibarotene were synthesized in 34-71% radiochemical yields.

Recurrent transient visual loss in a middle aged woman.

Visual loss is a common presenting complaint in primary care. We present a case of recurrent transient visual loss in a middle aged woman. Her funduscopy showed bilateral optic disc swelling. We have highlighted the differentiation of bilateral optic disc swelling at the primary care level as the management differs according to the diagnosis.

Simultaneous uncoupled expression and purification of the Dengue virus NS3 protease and NS2B co-factor domain.

Dengue Virus (DENV) infection is responsible for the world's most significant insect-borne viral disease. Despite an increasing global impact, there are neither prophylactic nor therapeutic options available for the effective treatment of DENV infection. An attractive target for antiviral drugs is the virally encoded trypsin-like serine protease (NS3pro) and its associated cofactor (NS2B). The NS2B-NS3pro complex is responsible for cleaving the viral polyprotein into separate functional viral proteins, and is therefore essential for replication. Recombinant expression of an active NS2B-NS3 protease has primarily been based on constructs linking the C-terminus of the approximately 40 amino acid hydrophilic cofactor domain of NS2B to the N-terminus of NS3pro via a flexible glycine linker. The resulting complex can be expressed in high yield, is soluble and catalytically active and has been used for most in vitro screening, inhibitor, and X-ray crystallographic studies over the last 15 years. Despite extensive analysis, no inhibitor drug candidates have been identified yet. Moreover, the effect of the artificial linker introduced between the protease and its cofactor is unknown. Two alternate methods for bacterial expression of non-covalently linked, catalytically active, NS2B-NS3pro complex are described here along with a comparison of the kinetics of substrate proteolysis and binding affinities of substrate-based aldehyde inhibitors. Both expression methods produced high yields of soluble protein with improved substrate proteolysis kinetics and inhibitor binding compared to their glycine-linked equivalent. The non-covalent association between NS2B and NS3pro is predicted to be more relevant for examining inhibitors that target cofactor-protease interactions rather than the protease active site. Furthermore, these approaches offer alternative strategies for the high yield co-expression of other protein assemblies.

Phosphor-Free Apple-White LEDs with Embedded Indium-Rich Nanostructures Grown on Strain Relaxed Nano-epitaxy GaN.

Phosphor-free apple-white light emitting diodes have been fabricated using a dual stacked InGaN/GaN multiple quantum wells comprising of a lower set of long wavelength emitting indium-rich nanostructures incorporated in multiple quantum wells with an upper set of cyan-green emitting multiple quantum wells. The light-emitting diodes were grown on nano-epitaxially lateral overgrown GaN template formed by regrowth of GaN over SiO(2) film patterned with an anodic aluminum oxide mask with holes of 125 nm diameter and a period of 250 nm. The growth of InGaN/GaN multiple quantum wells on these stress relaxed low defect density templates improves the internal quantum efficiency by 15% for the cyan-green multiple quantum wells. Higher emission intensity with redshift in the PL peak emission wavelength is obtained for the indium-rich nanostructures incorporated in multiple quantum wells. The quantum wells grown on the nano-epitaxially lateral overgrown GaN has a weaker piezoelectric field and hence shows a minimal peak shift with application of higher injection current. An enhancement of external quantum efficiency is achieved for the apple-white light emitting diodes grown on the nano-epitaxially lateral overgrown GaN template based on the light -output power measurement. The improvement in light extraction efficiency, η(extraction,) was found to be 34% for the cyan-green emission peak and 15% from the broad long wavelength emission with optimized lattice period.

Sprouty proteins: modified modulators, matchmakers or missing links?

Sprouty proteins are involved in organogenesis, particularly during the branching of endothelial tubes, and existing evidence suggests that Sprouty's point of action lies downstream of receptor signaling to inhibit the activation of the central Ras/Erk pathway. How Sprouty proteins accomplish their inhibitory action and whether they interact with other signaling pathways are significant questions. Sprouty proteins are devoid of any recognizable protein interaction domain, and clues as to how they function have been mainly derived from screening for interacting partners. Conserved across all the Sprouty proteins are three sequences: a Cbl-tyrosine kinase-binding (TKB) binding motif centered on an obligatorily phosphorylated tyrosine (Y55 in Sprouty2), a serine-rich motif (SRM) and a cysteine-rich domain (CRD). With the exception of a handful of proteins that bind to the N-terminus, most of the binding to Sprouty occurs via the CRD, predominantly by serine/threonine kinases that target sites within the SRM on Sprouty. Some of the resultant increase in phosphorylation is opposed by activated protein phosphatase 2A that binds to the N-terminal Cbl-TKB binding motif. Significantly, two ubiquitin E3 ligases also bind to the N-terminus of Sprouty: c-Cbl binds with high affinity to the TKB binding motif and SIAH2 binds constitutively to a different site; both proteins are able to direct the ubiquitination of Sprouty proteins and its destruction. The collective evidence points to Sprouty proteins as being substantially covalently-modified to control its location, stability, association, and destruction. With such stringent control of the Sproutys, the main question is what key proteins does this facilitator bring together?

Identification and characterization of mErk5-T, a novel Erk5/Bmk1 splice variant.

Extracellular regulated kinase 5 (ERK5) is an unusually large member of the MAP kinase family of signaling molecules that plays an important role in cellular proliferation, differentiation and survival. Recently, three transcriptional variants of murine Erk5 were described (mErk5-a, -b and -c) that result from alternate splicing across introns 1 and/or 2, the net effect of which is translation of a peptide that lacks the kinase domain. It has been demonstrated that expression of mErk5-b and -c impinge on the function of the full length mErk5 protein product via a dominant negative effect. Here, we report the identification of another murine Erk5 splice variant and the orthologous human transcript that arise due to alternate splicing of intron 4. Failure to splice out intron 4 introduces a premature in-frame stop codon that directs translation of a peptide lacking the nuclear localization signal (NLS) and proline-rich region (PR). Experimental characterization demonstrated that like mERK5, mERK5-T becomes phosphorylated by co-expression with a constitutively active mMEK5 (mMEK5DD), and is able to coimmunoprecipitate with both itself and mERK5. Unlike mERK5, however, activated ERK5-T is unable to translocate from the cytoplasm to the nucleus in HeLaS3 cells, causing the retention of active mERK5 in the cytoplasm. Taken together with previous reports of domain content modification of ERK5 via alternate splicing, these observations add to the suggestion that regulation of ERK5 signaling may be mediated, at least in part, at the level of RNA processing.

Ultrafast and large third-order nonlinear optical properties of CdS nanocrystals in polymeric film.

We report the ultrafast and large third-order nonlinear optical properties of CdS nanocrystals (NCs) embedded in a polymeric film. The CdS NCs of 2 nm radius are synthesized by an ion-exchange method and highly concentrated in the two layers near the surfaces of the polymeric film. The two-photon absorption coefficient and the optical Kerr coefficient are measured with laser pulses of 250 fs duration at 800 nm wavelength. The one-photon and two-photon figures of merit are determined to be 3.1 and 1.3, respectively, at irradiance of 2 GW/cm(2). The observed nonlinearities have a recovery time of approximately 1 ps. The two-photon-generated free carrier effects have also been observed and discussed. These results demonstrate that CdS NCs embedded in polymeric film are a promising candidate for optical switching applications.

Characterization and intraspinal grafting of EGF/bFGF-dependent neurospheres derived from embryonic rat spinal cord.

Recent advances in the isolation and characterization of neural precursor cells suggest that they have properties that would make them useful transplants for the treatment of central nervous system disorders. We demonstrate here that spinal cord cells isolated from embryonic day 14 Sprague-Dawley and Fischer 344 rats possess characteristics of precursor cells. They proliferate as undifferentiated neurospheres in the presence of EGF and bFGF and can be maintained in vitro or frozen, expanded and induced to differentiate into both neurons and glia. Exposure of these cells to serum in the absence of EGF and bFGF promotes differentiation into astrocytes; treatment with retinoic acid promotes differentiation into neurons. Spinal cord cells labeled with a nuclear dye or a recombinant adenovirus vector carrying the lacZ gene survive grafting into the injured spinal cord of immunosuppressed Sprague-Dawley rats and non-immunosuppressed Fischer 344 rats for up to 4 months following transplantation. In the presence of exogenously supplied BDNF, the grafted cells differentiate into both neurons and glia. These spinal cord cell grafts are permissive for growth by several populations of host axons, especially when combined with exogenous BDNF administration, as demonstrated by penetration into the graft of axons immunopositive for 5-HT and CGRP. Thus, precursor cells isolated from the embryonic spinal cord of rats, expanded in culture and genetically modified, are a promising type of transplant for repair of the injured spinal cord.

Intraspinal delivery of neurotrophin-3 using neural stem cells genetically modified by recombinant retrovirus.

Neural stem cells have been shown to participate in the repair of experimental CNS disorders. To examine their potential in spinal cord repair, we used retroviral vectors to genetically modify a clone of neural stem cells, C17, to overproduce neurotrophin-3 (NT-3). The cells were infected with a retrovirus construct containing the NT-3.IRES.lacZ/neo sequence and cloned by limiting dilution and selection for lacZ expression. We studied the characteristics of the modified neural stem cells in vitro and after transplantation into the intact spinal cord of immunosuppressed adult rats. Our results show that: (i) most of the genetically modified cells express both NT-3 and lacZ genes with a high coexpression ratio in vitro and after transplantation; and (ii) large numbers of the xenografted cells survive in the spinal cord of adult rats for at least 2 months, differentiate into neuronal and glial phenotypes, and migrate for long distances. We conclude that genetically modified neural stem cells, acting as a source of neurotrophic factors, have the potential to participate in spinal cord repair.

Transplants of fibroblasts genetically modified to express BDNF promote regeneration of adult rat rubrospinal axons and recovery of forelimb function.

Adult mammalian CNS neurons do not normally regenerate their severed axons. This failure has been attributed to scar tissue and inhibitory molecules at the injury site that block the regenerating axons, a lack of trophic support for the axotomized neurons, and intrinsic neuronal changes that follow axotomy, including cell atrophy and death. We studied whether transplants of fibroblasts genetically engineered to produce brain-derived neurotrophic factor (BDNF) would promote rubrospinal tract (RST) regeneration in adult rats. Primary fibroblasts were modified by retroviral-mediated transfer of a DNA construct encoding the human BDNF gene, an internal ribosomal entry site, and a fusion gene of lacZ and neomycin resistance genes. The modified fibroblasts produce biologically active BDNF in vitro. These cells were grafted into a partial cervical hemisection cavity that completely interrupted one RST. One and two months after lesion and transplantation, RST regeneration was demonstrated with retrograde and anterograde tracing techniques. Retrograde tracing with fluorogold showed that approximately 7% of RST neurons regenerated axons at least three to four segments caudal to the transplants. Anterograde tracing with biotinylated dextran amine revealed that the RST axons regenerated through and around the transplants, grew for long distances within white matter caudal to the transplant, and terminated in spinal cord gray matter regions that are the normal targets of RST axons. Transplants of unmodified primary fibroblasts or Gelfoam alone did not elicit regeneration. Behavioral tests demonstrated that recipients of BDNF-producing fibroblasts showed significant recovery of forelimb usage, which was abolished by a second lesion that transected the regenerated axons.

Intraspinal grafting of fibroblasts genetically modified by recombinant adenoviruses.

Intracerebral or intraspinal grafting of genetically modified primary fibroblasts has been shown to enhance functional recovery in several models of CNS disease, including spinal cord injury. Most of these studies utilized retrovirus vectors. In this report, we describe in vitro conditions for genetically modifying primary fibroblasts with recombinant adenovirus vectors carrying the lacZ or green fluorescent protein (GFP) genes. As intraspinal allografts in animals immunosuppressed by cyclosporin A, the genetically modified cells survived and expressed the transgenes for at least 2 months. We conclude that recombinant adenovirus vectors are efficient and convenient tools for ex vivo gene therapy in the CNS.

Application of recombinant adenovirus for in vivo gene delivery to spinal cord.

One strategy for treating spinal cord injury is to supply damaged neurons with the appropriate neurotrophins either by direct delivery or by transfer of the corresponding genes using viral vectors. Here we report the feasibility of using recombinant adenovirus for in vivo gene transfer in spinal cord. After injection of a recombinant adenovirus carrying a beta-galactosidase (beta-gal) reporter gene into the mid-thoracic spinal cord of adult rats, transgene expression occurred not only in several types of cells around the injection site but also in neurons whose axons project to this region from rostral or caudal to the injection site. Among labeled neurons were those of the red nucleus, the vestibular nuclei, reticular formation, locus coeruleus, and Clarke's nucleus. A non-specific immune reaction, which could be blocked by immunosuppression with Cyclosporin A, reduced the number of transduced cells surviving at the injection site by 1 month. In neurons away from the injection site, where the immune response was minimal, transgene expression lasted for at least 2 months. These results support the idea that recombinant adenovirus can be used in the spinal cord for in vivo delivery of therapeutic genes important for supporting neuron survival and axon regeneration.

Brain oxytocin receptor antagonism disinhibits sodium appetite in preweanling rats.

Previous studies have shown that preweanling rats do not express an endogenous sodium appetite until postnatal day 12. The present studies tested the hypothesis that prior to 12 days of age sodium appetite, induced by either central administration of angiotensin II (AngII) or adrenalectomy, is inhibited by endogenous oxytocin (OT). After 9- or 10-day old animals were given a central injection of either an OT receptor antagonist or vehicle, they were infused intraorally with 4% sodium chloride which the animals could either swallow or reject. Intake was measured as the increase from initial body weight. There was very little sodium consumption by vehicle-injected animals that received sham surgery or adrenalectomy; however, the OT receptor antagonist significantly elevated sodium consumption in adrenalectomized animals. The OT antagonist also potentiated sodium intake after AngII pretreatment. These results suggest that the neurochemical circuits necessary for the expression of sodium appetite are present and functional as early as postnatal day 9; however, until 12 days of age this behavior is suppressed by endogenous OT.

Blockade of central angiotensin II type 1 and type 2 receptors suppresses adrenalectomy-induced NaCl intake in rats.

Removal of the adrenal glands, the main site for the synthesis of aldosterone, produces an intake of sodium that is essential for survival. Using central blockade of angiotensin II (Ang II) receptors with SarIle Ang II, previous studies have shown that this intake depends on the stimulation of the brain angiotensin system. In the present study, using intracerebroventricular injection of specific antagonists of Ang II type 1 (AT1) or type 2 (AT2) receptors (losartan and PD 123319, respectively), we confirm that activation of brain angiotensin is essential for the expression of adrenalectomy-induced NaCl intake. Moreover, we show that (a) AT1 but not AT2 receptor blockade alone suppresses NaCl intake and (b) doses of AT1 and AT2 receptor antagonists that separately have no effect on NaCl intake, suppress the behavior when combined. It is proposed that AT1 receptors mediate the natriorexigenic effect of Ang II and that AT2 receptors have a permissive role on AT1 receptor stimulation.

Electrophysiological studies on acute tetrodotoxin poisoning: a case report.

Tetrodotoxin (TTN) is a marine neurotoxin that is sometimes responsible for the mortality of pufferfish consumers. A case of human tetrodotoxication caused by ingestion of Goby is reported. The patient developed perioral numbness, acroparesthesia, generalized weakness and respiratory failure within hours after ingestion. The electrophysiological studies at the 16th hour of intoxication revealed generalized sensorimotor axonopathy, but no disturbance on the neuromuscular junction motor endplate. Human electrophysiological studies of acute TTX intoxication have rarely been documented in literature.

Effects of prostaglandin E2 and F2 alpha on the skeleton of osteopenic ovariectomized rats.

This article contains the histomorphometric evaluation of the effects of prostaglandin F2 alpha (PGF2 alpha) on cancellous bone from the lumbar vertebra and cortical bone from the tibial shaft of ovariectomized, osteopenic rats. These effects were then compared with those of prostaglandin E2 (PGE2). Three-month-old rats were either ovariectomized (ovx) or sham-ovx. Then, either PGF2 alpha or PGE2 in doses of 1 and 3 mg/kg/day was given subcutaneously for 21 days at 150 days post ovx. Histomorphometric analysis was performed separately on both the primary and secondary spongiosae of the fourth lumbar vertebral bodies (LVB) and on tibial shafts. The ovx rats exhibited osteopenia in both primary (-23% to -37%) and secondary (-20%) spongiosae of the LVB, but not in the tibial shafts at 150 and 171 days post ovx. In the LVB, PGE2 in doses of 1 or 3 mg/kg/day for 21 days restored trabecular bone volume to the levels of sham-ovx controls in the primary spongiosa. However, in the secondary spongiosa, the treatments only thickened the trabeculae. The effects of the PGF2 alpha treatment were similar to those of the PGE2 in both the primary and the secondary spongiosae. While both PGF2 alpha and PGE2 treatments stimulated bone formation in the LVB as indicated by the increases in labeled perimeter, tissue and bone area-based bone formation rates, PGE2 is about 10 times more potent than PGF2 alpha in these effects. The PGE2 treatment also elevated activation frequency in the LVB, while the PGF2 alpha treatment did not. The treatments differed in that PGE2 at these dose levels did not alter the eroded surface in the LVB while PGF2 alpha decreased it significantly. Thus, the increase of the ratio of labeled to eroded perimeter in the LVB in PGF2 alpha-treated animals was much more than that in PGE2-treated animals. In the tibial shafts, PGE2 in doses of 1 and 3 mg/kg/day produced new marrow trabeculae in 2 of 6 and 3 of 6 of the ovx rats. However, no new trabecula was found in PGF2 alpha-treated tibial shafts. Higher doses of PGE2 also increased periosteal labeled perimeter, MAR, and BFR/BS, while PGF2 alpha did not produce any significant change in these parameters. Both PGE2 and PGF2 alpha in doses of 1 and 3 mg/kg/day increased the labeled perimeter, MAR and BFR/BS and decreased the eroded perimeter in the endocortical surface. We concluded that both PGF2 alpha and PGE2 in doses of 1 and 3 mg/kg/day for 21 days exhibited anabolic bone effects. The effects were mostly confined to an increase in trabecular volume in the primary spongiosa of the LVB and in the endocortical surface of tibial shafts. The tissue level mechanism behind this appears to be that PGE2 and PGF2 alpha can both stimulate osteoblast recruitment and activity. Overall, we found PGE2 to be more potent than PGF2 alpha at the same dose level at the endocortical surface. Furthermore, new marrow trabecular bone formed only after PGE2 treatment. PGF2 alpha differed from PGE2 by significantly reducing the trabecular eroded surface in ovx rats.

Maintenance therapy for added bone mass or how to keep the profit after withdrawal of therapy of osteopenia.

Since continuous therapy for osteoporosis can be expensive and may have detrimental effects, there is a need to develop a strategy to maintain bone mass after withdrawal of treatment. The bone maintained by estrogen and calcitonin therapies and exercise, but the added bone induced by anabolic agents disappears upon cessation of treatment. To avoid this pitfall, the concepts of activation, restore and maintain (ARM) or loss, restore and maintain (LRM), the on/off administration of combined anabolic agent with an antiresorptive or antiactivation agent, and cyclical treatment of the two regimes have been employed successfully in "keeping the profit" (maintaining bone) in preclinical studies. The data for the disappearance of bone upon cessation of certain osteopenic treatments, its mechanism of loss and the development of maintenance concept and subsequent preclinical studies indicate that there was no need for costly continuous therapy in the treatment strategy for osteoporosis.

Effects of glutamate, N-methyl-D-aspartate, high potassium, and hypoxia on unit discharges in CA1 area of hippocampal slices of DBA and C57 mice.

We studied effects of L-glutamate, N-methyl-D-aspartate (NMDA), high K+, and hypoxia on spontaneous unit discharges in stratum pyramidale of CA1 region of hippocampal slices in DBA and C57 mice aged 3-4 and 5-6 weeks. Application of L-glutamate (0.5-2.0 mM), NMDA (5-20 microM), high K+ (8.5 mM), and a brief period of hypoxia (1 min) to the perfused artificial cerebrospinal fluid (ACSF) all produced different degrees of spontaneous high-frequency discharges from CA1 area of hippocampal slices of both DBA and C57 mice. Two types of responses recorded extracellularly occurred after these manipulations: high-frequency repetitive single spikes and bursts of multiple population spikes. The rate and type of responses from CA1 region of hippocampal slices after these manipulations were different and depended on the strain and age of mice and the nature of manipulations. In general, hippocampal slices from audiogenic seizure-susceptible DBA mice were more sensitive than those from audiogenic seizure-resistant C57 mice, and hippocampal slices from younger animals were more susceptible than those from older ones. Thus, DBA mice aged 3-4 weeks of age were most susceptible and C57 mice aged 5-6 weeks were least susceptible to all these pharmacological, ionic, and hypoxic manipulations. Bursts of multiple population spikes were the most common responses in DBA mice and in younger animals, and repetitive single spikes were the predominant responses in C57 mice and in older animals. In all groups of animals, the average spontaneous discharge rate was highest after L-glutamate perfusion, next highest after NMDA, and lowest after high K+ and hypoxia. The latency of the appearance of spontaneous epileptiform activity from CA1 region of hippocampal slices was long (> 2 min) after NMDA perfusion and short (< 1 min) after L-glutamate, high K+ and hypoxia. The duration of the increased spontaneous discharges was short (-1 min) after L-glutamate perfusion, long (> 3 min) after high K+ and hypoxia, and between short and long after NMDA perfusion. These results suggest that age and strain of animal and nature of stimulus precipitate different patterns of epileptiform activity in CNS.

Epilepsy and intracranial meningiomas.

BACKGROUND: Only a few studies have reflected the incidence and causes of preoperative and postoperative seizures in meningiomas. This study concerned the incidence and types of preoperative epilepsy, and the predisposing factors for postoperative epilepsy in meningiomas. METHODS: Epilepsy occurred in 323 surgically treated intracranial meningiomas. The focus here is different types of seizures, tumor locations, onset of seizures and the histopathological features of the meningiomas. RESULTS: From analysis of 323 patients with intracranial meningiomas, aged 10 to 79, 98 (30.3%) were found to have different types of preoperative epilepsy; in 32 (32.7%) of them, the seizures persisted postoperatively. Among 225 patients without preoperative seizures, 39 (17.3%) developed postoperative seizures. Thus, a history of preoperative seizures is a significant index (p < 0.005) for predicting the occurrence of postoperative seizures. In a total of 71 patients with postoperative seizures, the precipitating factors in the first week were cerebral edema and hemorrhage at the surgical sites. In late postoperative seizures (onset beyond one week post-surgery), the main cause was tumor recurrence. Patients with sagittal and convexity meningiomas had a higher incidence of seizures. There is no relationship between the histopathological features of the tumor and the occurrence of epilepsy in meningiomas. CONCLUSIONS: There is a significant incidence of postoperative seizures in meningioma patients with a history of preoperative seizures. Surgical excision of tumor, absence of postoperative hemorrhage or edema and anticonvulsant therapy reduced the occurrence of postoperative seizures.

Effect of kainic acid on unit discharge in CA1 area of hippocampal slice of DBA and C57 mice.

Spontaneous unit discharges in stratum pyramidale of CA1 area of hippocampal slices from DBA and C57 mice at different ages were recorded extracellularly. The average rate and amplitude of the spontaneous discharges from CA1 area of hippocampal slices bathed in artificial cerebrospinal fluid (aCSF) were not different between DBA and C57 mice at either 3-4 or 5-6 weeks of age. Bath application of kainic acid (KA) in concentrations of 0.5-1.0 microM produced different responses in CA1 area from these two strains of mice. In DBA mice at age 3-4 weeks, when they are most susceptible to audiogenic seizures, KA perfusion induced high-frequency repetitive single spikes and bursts of multiple population spikes in CA1 area. Very high-frequency discharges (10-fold higher than most responses) were also observed in 20% of all slices. In audiogenic seizure resistant C57 mice at age 3-4 weeks, KA perfusion at the same doses induced only the repetitive single spikes. The rate of spontaneous discharges was much lower than that in DBA mice. No burst of multiple population spikes nor very high-frequency responses were recorded in C57 mice. At age 5-6 weeks, when both DBA and C57 mice are resistant to audiogenic seizures, the rate of spontaneous discharges recorded from the CA1 area during and after KA perfusion was lower than that at age 3-4 weeks, and there was no significant difference between DBA and C57 mice.(ABSTRACT TRUNCATED AT 250 WORDS)