Interaction of Arc with CaM kinase II and stimulation of neurite extension by Arc in neuroblastoma cells expressing CaM kinase II

We investigated the relationship between Arc (activity-regulated cytoskeleton-associated protein) and Ca(2+)/calmodulin-dependent protein kinase II (CaM kinase II). Arc and CaM kinase II were concentrated in the postsynaptic density. These proteins were accumulated after electroconvulsive treatment. Arc increased about 2.5-fold within 30 min and was maintained at this level for 8h after the stimulation. CaM kinase II also increased within 30 min and remained at this level for at least 24h. The interaction of Arc with CaM kinase II was demonstrated using GST-Arc fusion protein, and confirmed in neuroblastoma cells by immunoprecipitation. We examined the function of Arc by introducing Arc cDNA into neuroblastoma cells expressing CaM kinase II. The cells expressing both Arc and CaM kinase II had longer neurites than those expressing CaM kinase II alone. Arc itself did not promote neurite outgrowth. The growth of neurites by Arc was completely blocked by treatment with KN62, an inhibitor of CaM kinases. These results indicated that Arc potentiated the action of CaM kinase II for neurite extension.     

Congenital aplasia of the extensor muscles of the fingers and thumb associated with generalized polyneuropathy: an autosomal recessive trait

Three sibs born to normal but consanguinous parents had flexion deformities of the thumb and fingers on one hand and sensory deficit in the other hand. Extensor muscles were absent or vestigial in all of them. Polyneuropathic electrophysiological findings were detected in 4 limbs. This is an apparently autosomal recessive trait previously unreported.     

Chorioamnionitis caused by Serratia marcescens in a non-immunocompromised host

A 26 year old pregnant woman with antithrombin III deficiency developed recurrent septicaemia with Serratia marcescens. In spite of the administration of antibiotics, high grade fever persisted. She subsequently manifested lower abdominal pain, and spontaneous abortion occurred. After the abortion, she became completely afebrile. The amnion was turbid, and microscopic examination of the placenta showed haemorrhage and massive infiltration of neutrophils, suggestive of infectious chorioamnionitis. Pulsed field gel electrophoresis showed that isolates from the blood, urine, and vaginal discharge were genetically identical. Intravenous pyelography revealed that she had a bilateral completed double ureter. It was thought that a urinary tract anomaly caused infection with S marcescens, and the pathogen spread to the chorioamnion via the bloodstream. This is the first report of chorioamnionitis caused by S marcescens in a non-immunocompromised host. In addition, these findings indicate that the chorioamnion can serve as a site for persistent infection in normal pregnancies.     

Soluble intercellular adhesion molecule-I levels in sera of patients with Kawasaki disease]

We investigated whether serum levels of soluble ICAM-1 antigens increases during acute Kawasaki disease (KD). We also compared levels in acute KD with those in anaphylactoid purpura (AP) and in measles. Serum soluble ICAM-1 levels were measured by a double determinant immunoassay using two monoclonal antibodies in the FAST system. Serum levels of tumor necrosis factor-alpha (TNF-alpha) were measured by a specific and sensitive sandwich enzyme immunoassay. Patients with KD, but not with AP or measles, had increased ICAM-1 levels in serum during the acute stage. In addition, during the acute stage, KD patients with coronary-artery lesions (CAL) were found to have increased ICAM-1 levels in serum compared to patients without CAL. We found a positive correlation between serum levels of ICAM-1 and levels of TNF-alpha during acute KD. Our results suggest that the serum level of soluble ICAM-1 is an important immunologic parameter for determining the severity of vascular damage during acute KD.     

Activation of the dorsal premotor cortex and pre-supplementary motor area of humans during an auditory conditional motor task

Using functional magnetic resonance imaging (fMRI), we measured regional blood flow to examine which motor areas of the human cerebral cortex are preferentially involved in an auditory conditional motor behavior. As a conditional motor task, randomly selected 330 or 660 Hz tones were presented to the subjects every 1. 0 s. The low and high tones indicated that the subjects should initiate three successive opposition movements by tapping together the right thumb and index finger or the right thumb and little finger, respectively. As a control task, the same subjects were asked to alternate the two opposition movements, in response to randomly selected tones that were presented at the same frequencies. Between the two tasks, MRI images were also scanned in the resting state while the tones were presented in the same way. Comparing the images during each of the two tasks with images during the resting state, it was observed that several frontal motor areas, including the primary motor cortex, dorsal premotor cortex (PMd), supplementary motor area (SMA), and pre-SMA, were activated. However, preferential activation during the conditional motor task was observed only in the PMd and pre-SMA of the subjects' left (contralateral) frontal cortex. The PMd has been thought to play an important role in transforming conditional as well as spatial visual cues into corresponding motor responses, but our results suggest that the PMd along with the pre-SMA are the sites where more general and extensive sensorimotor integration takes place.     

The properties of the Kir6.1–6.2 tandem channel co-expressed with SUR2A

Functional ATP-sensitive K (KATP) channels have an octameric subunit structure with four pore-forming subunits (Kir6.x) and four sulfonylurea receptors (SURx). In the present study, the properties of the heteromeric KATP channel whose pore subunits are composed of Kir6.1 and Kir6.2 were examined using a heterologous expression system. In COS7 cells co-transfected with Kir6.1, Kir6.2 and SUR2A at a ratio of 1:1:2, KATP channels showed various unitary conductances between those of Kir6.1/SUR2A (33.6+/-4.2 pS) and Kir6.2/ SUR2A (67.1+/-1.6 pS). Kir6.1-6.2 tandem protein, constructed by fusing the C-terminus of Kir6.1 to the N-terminus of Kir6.2 with a ten glutamine linker sequence, also formed a channel with an intermediate conductance (58.9+/-1.5 pS). Kir6.2 and Kir6.1-6.2 showed similar sensitivity to ATP4-: half-maximal inhibition (IC50) was obtained at 14.1+/-12.8 microM and 17.6+/-9.6 microM, respectively. In the presence of Mg2+, Kir6. 1-6.2 was significantly less sensitive than Kir6.2 to MgATP (IC50=95.5+/-49.6 microM versus 18.9+/-5.0 microM). These results suggest that Kir6.1 and Kir6.2 are endowed with the potential to form a heteromeric KATP channel, which has a low sensitivity to MgATP.     

Effects of recombinant leukocyte interferon on serum immunoglobulin concentrations and lymphocyte subpopulations in chronic hepatitis B

To investigate immune effects of interferon (IFN) therapy in hepatitis B e antigen (HBeAg)-positive chronic hepatitis B, serum immunoglobulin concentrations and peripheral lymphocyte subpopulations were sequentially studied before, during, and after therapy in nine patients who were treated with recombinant human -IFN in doses ranging from 3 to 10 million units per day for 28 days. Serum immunoglobulin A levels decreased significantly, from 414±23 mg/dl (mean ± SE) to 379±28 mg/dl (P<0.05), after the first week of therapy and to a bottom value of 323±20 mg/dl (P<0.001) at the fourth week. Immunoglobulin G levels decreased significantly, from 2603±175 to 2328±169 mg/dl (P<0.005), after the first week of therapy and to a bottom value of 2005±199 mg/dl (P<0.001) at the fourth week. Immunoglobulin M levels were also reduced significantly after 3 weeks of therapy (from 229±23 to 188±15 mg/dl;P<0.01). These reductions in immunoglobulins A, G, and M returned to pretreatment levels by 4 months after the end of the therapy. In lymphocyte subpopulations, significant depressions were found in CD3-, CD4-, CD8-, and B1-positive cells in peripheral blood after the first week of therapy (CD3, from 1700±114 to 1234±114/mm³,P<0.005; CD4, from 1036±88 to 780±64/mm³,P<0.005; CD8, from 620±57 to 426±60/mm³,P<0.05; and B1, from 519±84 to 276±48/mm³,P<0.01) followed during therapy, while Leul la-positive cells did not change significantly. During the 6-month follow-up period, three patients had a sustained clinical remission in which HBeAg disappeared from serum. Disappearance of HBeAg was unassociated with initial levels or percentage changes of serum immunoglobulins and peripheral lymphocytes expressing each of the test markers in these patients. These findings suggest that immune effects of IFN therapy are independent from its antiviral effects.     

A case of alcoholic cirrhosis demonstrating long-term poor-visualization of the hepatic image on 99mTc-phytate scintigraphy

A 27-year-old female patient with alcoholic cirrhosis was reported. She was admitted to the hospital because of jaundice and ascites after heavy drinking. She had a history of drinking Japanese Sake in quantities of more than 5 go/day (900 ml/day) for 7 years. On admission, she was icteric, and had both hepatosplenomegaly and ascites. Laboratory data showed an elevation of serum transaminase and bilirubin, and a decrease in the albumin and prothrombin values. A biopsy specimen of the liver showed pericellular fibrosis, fatty change, Mallory bodies and regenerative nodules, and revealed findings compatible with alcoholic cirrhosis. A 99mTc-N-pyridoxyl-5-methyltryptophan scintigram showed hepatomegaly. On the 99mTc-phytate scintigram, the uptake of radioisotope to the liver was markedly decreased with the increased uptake to the spleen and bone marrow. Even 6 months after the onset, poor visualization of the hepatic image on 99mTc-phytate scintigram continued. This is the first report of alcoholic cirrhosis demonstrating a long-term poor visualization of the hepatic image on 99mTc-phytate scintigraphy.     

In vitro hydrolysis of blends from enantiomeric poly(lactide)s. Part 4: well-homo-crystallized blend and nonblended films

Well-homocrystallized enantiomeric blend and nonblended films were prepared from poly(L-lactide), i.e., poly(L-lactic acid) (PLLA) and poly(D-lactide), i.e., poly(D-lactic acid) (PDLA) by crystallization from the melt. The effects of enantiomeric blending on the in vitro autocatalytic hydrolysis of the homo-crystallized polylactide, i.e., poly(lactic acid) (PLA) films were investigated in phosphate-buffered solution (pH 7.4) at 37 degrees C for up to 24 months. In the period of 0-12 months, the effects of enantiomeric polymer blending on the autocatalytic hydrolysis were very small. This finding reflects that in the PLLA/PDLA blend film separate homo-crystallization of PLLA and PDLA into the respective crystallites reduced the peculiar strong interaction between PLLA and PDLA chains in the amorphous region between the homo-crystalline regions. In the period of 12-24 months, enantiomeric polymer blending significantly retarded the autocatalytic hydrolysis of the PLLA/PDLA blend film compared with that of the nonblended PLLA and PDLA films. This is attributable to the increased chain mobility and the reduced entanglement effects due to the chain cleavage to a great extent, resulting in the enhanced interaction between PLLA and PDLA chains. It was also revealed that the hydrolyzabilities of the PLA films can be widely varied by enantiomeric polymer blending, crystalline species and their amounts, and molecular weight.