Intracellular and virion 35 S RNA species of murine sarcoma and leukemia viruses

RNA molecules from the following sources were treated with dimethyl sulfoxide to dissociate noncovalent aggregates and resolved by electrophoresis on polyacrylamide gels: (1) transformed MSV(MLV)-producing Balb/3T3 cells (MSV-39, clone 24), (2) transformed MSV(MLV)-producing rat cells (78A1), (3) MSV transformed, non virus-producing hamster cells (HT-1), (4) MLV-producing Balb/3T3 cells, (5) MLV-producing NIH/3T3 cells (6) 60–70 S RNA from MSV(MLV) virions, and (7) 60–70 S RNA from MLV virions. Virus-specific RNA was detected by hybridization of RNA in gel fractions with the ³H-DNA product of the MSV(MLV) RNA-directed DNA polymerase. Two well defined viral RNA species with sedimentation coefficients of 35 S and 20 S, but none of intermediate size, were detected in both MSV(MLV) producing cell lines. The non virus-producing HT-1 cell line contained a viral RNA species slightly smaller than 35 S, about 33 S, but no detectable 20 S virus-specific RNA. These results with 78A1 and HT-1 cells agree with previous conclusions based on rate-zonal centrifugation in sucrose density gradients (Tsuchida et al., 1972). The two MLV-producing mouse cell lines contained a well defined 35 S RNA peak, a somewhat less pronounced 20 S RNA peak, and heterogeneous RNA species of smaller molecular weights. Although both 35 S and 20 S virus RNA species were detected in cells replicating murine oncornaviruses, labeled 60–70 S RNA isolated from MSV(MLV) virions (consisting mainly of MLV) and MLV virions, and dissociated with dimethyl sulfoxide or by heat, consisted of 35 S RNA and about 10–15% 7 S RNA plus 4–5 RNA, but no detectable 20 S RNA. Hybridization-competition experiments using MSV(MLV) 60–70 S ³H-RNA (consisting mainly of MLV RNA) and saturating amounts of the unlabeled DNA product of the MSV(MLV) RNA-directed DNA polymerase showed that 35 S RNA from cells replicating MSV(MLV) shares virtually all of its nucleotide sequences with MSV(MLV) 70 S RNA; these data suggest that intracellular 35 S RNA species are the major precursors to virion 70 S RNA. In contrast, 33 S RNA from HT-1 cells shares only about 50% of its sequences with MSV(MLV) 70 S RNA indicating that only part of the sequences of MSV(MLV) 60–70 S RNA are integrated and/or transcribed in this non virus-producing MSV transformed cell.     

Expression of Functional Molecules in Non-Hodgkin's Lymphoma

It is well known that non Hodgkin's lymphoma (NHL) cells express various antigens which are normally involved in a variety of functions. In addition, NHL is diverse in its proliferative capacity. To investigate the relation between these factors and the clinical picture, 45 cases of NHL were studied by immunohistochemistry using snap-frozen materials obtained before therapy. Reactivities with 27 monoclonal antibodies were examined and the results were correlated with clinical findings. The expression of surface μ and CAM-1 in B-NHLs and CD25 in T-NHLs were significantly associated with bone marrow involvement. B-NHLs without expression of CD21(B2) and T-NHLs with CD25 were seen more frequently in cases with a LDH value of over 500 units/ml. The positivity rate of Ki-67 on B-NHLs was correlated with serum LDH value, NHL histologic classification, and overall survival. These data indicate that immunophenotyping and determination of the proliferative capacity of NHL are of value not only for confirmation of the histopathologic classification of the tumor but also for assessment of clinical behavior.     

A novel translocation involving chromosomes 2, 9, 14, and 22 in chronic myeloid leukemia

A 46-year-old man with chronic myelogenous leukemia was found to have a new complex translocation. In chronic phase, all of the bone marrow cells had a rearrangement of a t(2;9;14;22) (p21;q34;q32;q11). Southern blot analysis of leukocyte DNA revealed rearrangement of the breakpoint cluster region (bcr) within the 5.8-Kb bcr. The patient eventually died in blast crisis 28 months later. The cytogenetic findings of bone marrow cells showed a 46,XY,t(2;9;14;22)(p21;q34;q32;qll),add(lp),del(3q) karyotype in blast crisis.     

Reduced NR2A expression and prolonged decay of NMDA receptor-mediated synaptic current in rat vagal motoneurons following axotomy

To elucidate characteristic changes in the N -methyl- d -aspartate (NMDA) receptor on neurons following axotomy, subunit expressions and functional features of the NMDA receptor were examined in the dorsal motor nucleus of vagus (DMV) of rats receiving vagal axotomy at the neck. Western blotting analysis demonstrated that the expression of NR2A decreased 2–3 days after in vivo axotomy, while expression of NR1 and NR2B, NR2C and NR2D subunits did not change significantly. To examine the functional changes, patch clamp recordings in whole-cell mode were employed on the axotomized DMV neurons identified by retrograde labelling with fluorescent dye. The amplitude ratios of ifenprodil-sensitive components of NMDA response and d , l -2-amino-5-phosphovaleric acid (APV)-sensitive evoked postsynaptic current increased after axotomy. In addition, APV-sensitive postsynaptic currents exhibited a longer decay time in identified axotomized vagal motoneurons than in control neurons. No significant differences in the current density of the NMDA response and the peak amplitude of APV-sensitive synaptic currents were observed between axotomized and intact DMV neurons. In conclusion, a decrease in NR2A expression results in the appearance of functional characteristics of the NMDA receptor predominantly containing the NR2B subunit. This might lead to a long-term increase of the susceptibility of neurons to excitotoxicity.     

A new balloon catheter for selective cerebral perfusion

Selective cerebral perfusion (SCP) is a better method than hypothermic circulatory arrest for brain protection in aortic arch surgery because of fewer time limitations; however; it is more troublesome during surgery. We developed a new catheter for SCP, the SP Stud catheter, with a rib-surface balloon to reduce slipping to eliminate the need for snaring or clamping. To evaluate the slipping resistance of the new catheter, we compared it with two different balloon catheters; a Foley balloon catheter with a smooth-surface balloon as long as the SP Stud catheter, and a Retro-TH catheter with a smooth-surface balloon of discoid shape half as long as the SP Stud catheter. The “draw-strength”, which refers to the slipping resistance, was measured in 20 autopsy carotid arteries in the three groups. The average draw-strengths of SP Stud catheter, foley balloon catheter, and Retro-TH catheter were 92.3±4.0, 20.8 ±1.5, and 17.9±0.8g, respectively, in sclerotic carotid arteries. The SP Stud catheter demonstrated approximately four times more resistance to slipping than the Foley balloon or Retro-TH catheters. The SP Stud catheter demonstrated a higher slipping resistance than conventional catheters, which may allow selective cerebral perfusion without snaring or clamping the carotid arteries, minimize the change of cerebral embolism, and clear the operative field.     

The action of BDNF on GABAA currents changes from potentiating to suppressing during maturation of rat hippocampal CA1 pyramidal neurons

During the development of the hippocampus, the action of GABA shifts from depolarizing to hyperpolarizing, and brain-derived neurotrophic factor (BDNF) has important roles in GABAergic transmission. We demonstrate that BDNF (20 ng ml⁻¹) rapidly and reversibly potentiates postsynaptic GABA_A receptor-mediated currents (by 80.5 ± 14.3 %, n = 10) in hippocampal CA1 pyramidal neurons isolated from postnatal day (P)6 rats, using nystatin-perforated patch-clamp recordings. This potentiation is caused by an elevation of intracellular Ca²⁺ that occurs in response to the activation of Trk B receptor tyrosine kinase and phospholipase C-γ. The modulation of the GABA_A responses by BDNF in hippocampal CA1 pyramidal neurons isolated from P10 rats was more diverse (from potentiating to inhibitory), and at P14, BDNF induced a long-lasting inhibition. In addition, Ca²⁺/calmodulin-dependent protein kinase 2 plays important roles in the potentiating, but not in the inhibitory effect, of BDNF on the GABA_A responses. These results suggest that changes in the intracellular signalling pathway could contribute to the developmental shift of the actions of BDNF on inhibitory systems.     

Cerebral hemodynamics in patients with moyamoya disease. A study of regional cerebral blood flow by the 133Xe inhalation method

Regional cerebral blood flow was measured by the 133Xe inhalation method in 20 young patients with moyamoya disease and five young healthy volunteers. Most patients showed low values of mean hemispheric blood flow in both hemispheres. Regional cerebral blood flow was at a low value in the upper frontal region and at an almost average value in the posterotemporal and occipital regions, which was different from the "hyperfrontal" pattern in healthy volunteers. Regional cerebral blood flow was reduced evenly by hyperventilation. By 5% CO2 inhalation, regional cerebral blood flow was increased in the temporooccipital regions and was nearly unchanged or decreased in the frontal region.     

Venodilator effects of adenosine triphosphate and sodium nitroprusside; Comparisons during controlled hypotension

Adenosine triphosphate as well as sodium nitroprusside has been used for hypotensive anesthesia. The purpose of this study was to examine the possibility that two hypotensive drugs may exert different effects on venous capacitance during controlled hypotension. In rats anesthetized with ketamine, mean arterial pressure was lowered to 50mmHg by intravenous infusion of adenosine triphosphate or sodium nitroprusside. Venous capacitance was assessed before and during induced hypotension by measuring the mean circulatory filling pressure (MCFP). MCFP was measured after briefly arresting the circulation by inflating an indwelling balloon in the right atrium. MCFP was lower during adenosine triphosphate-induced as well as sodium nitroprusside-induced hypotension as compared with the respective value at control (P < 0.01 for adenosine triphosphate and sodium nitroprusside). However, the decrease in MCFP by adenosine triphosphate (0.8 ± 0.1mmHg) was less (P < 0.01) than that by sodium nitroprusside (2.3 ± 0.3mmHg). These results suggest that at a comparable level of arterial hypotension venodilator effect of adenosine triphosphate was less than that of sodium nitroprusside. Less venodilatation during adenosine triphosphate-induced hypotension may contribute to the maintenance of cardiac output during hypotensive anesthesia.(Hoka S, Takeshita A, Aishima K et al.: Venodilator effects of adenosine triphosphate and sodium nitroprusside; comparisons during controlled hypotension. J Anesth 1: 144–147, 1987)     

Carpal Tunnel Syndrome Due to Iatrogenic Amyloidosis After Domino Liver Transplantation From Hereditary Transthyretin Amyloidosis: A Case Report

BackgroundCarpal tunnel syndrome is the most common compression syndrome of the peripheral nerve. Transthyretin amyloidosis and dialysis-related β2-microglobulin amyloidosis are known causes of carpal tunnel syndrome.Case ReportA Japanese woman showed carpal tunnel syndrome 16 years after a domino liver transplantation (DLT) from the donor with hereditary transthyretin amyloidosis. DLT indication was congenital extrahepatic portosystemic shunt, and the patient had been put on maintenance hemodialysis because of chronic kidney disease 6 years before DLT. Moreover, the amyloid precursor protein of the patient was histologically confirmed not to be β2-microglobulin, but transthyretin.ConclusionsThe existence of amyloid was speculated when the patient who underwent DLT from hereditary transthyretin amyloidosis showed carpal tunnel syndrome. Additionally, elucidating the amyloid precursor protein when the patient has another cause of amyloidosis is necessary.     

Long-term Natural Course of the Osteochondral Lesion of the Talus in a Child: A Case Report

Recent reports have described midterm natural courses of osteochondral lesion of the talus (OLT) and lack of progression of ankle osteoarthritis (OA) in adult patients. The relationship between the OLT managed with nonoperative treatment and development of OA in children remains unknown. We report the long-term course of medial OLT in a 12-year-old female who was treated nonoperatively for 10 years. Radiographically, no osteoarthritic changes were observed at the first examination. She initially returned to her basketball club after nonoperative treatment. Although daily activities were not restricted, limitation of recreational activities began to appear at 4 years of follow-up. Subsequently, plain radiographs revealed bone absorption around the osteochondral fragment and osteophyte formation at the medial gutter, then ankle OA was advanced at the final follow-up.