An autoradiographic study of binding of iodinated spider toxin to lobster muscle

Distribution of the binding sites of Joro spider toxin (JSTX), a specific inhibitor of the glutamate receptors in the crustacean neuromuscular synapse, was studied by using autoradiography. JSTX was synthesized and made radioactive by conjugation with iodine-125. 125I-JSTX irreversibly blocked the excitatory postsynaptic potentials of the lobster neuromuscular synapse in a similar manner as the natural spider toxin. Light microscopic autoradiography of 125I-JSTX treated muscle showed sporadic aggregates of reduced silver grains on the surface of muscles. Electron microscopy of adjoining ultrathin sections revealed that these spots corresponded to the fraction of sarcolemma apposed to axonal terminals with or without synaptic junctional profiles. This finding gives morphological support to the formulation that JSTX binds to the glutamate receptor-ion channel molecules.     

Human herpesvirus 6 downregulates major histocompatibility complex class I in dendritic cells

The expression of major histocompatibility complex (MHC) class I, class II, CD1a, and CD 83 in dendritic cells (DCs) after infection with human herpesvirus 6 (HHV-6) was examined. Whereas there was no significant change in the expression of CD1a, CD83, and MHC class II in infected DCs, MHC class I expression was downregulated after infection with HHV-6 variant A but not HHV-6B. The expression of HHV-6 immediate-early or early genes was required for the downregulation of MHC class I. The de novo synthesis of MHC class I was greatly suppressed by infection with HHV-6A in DCs, while its rate of degradation was only slightly elevated. These results suggest that HHV-6A may escape from the host immune system in DCs by causing the downregulation of MHC class I synthesis.     

Monoclonal antibodies to monkey brain choline acetyltransferase: production and immunohistochemistry

Two monoclonal antibodies to monkey brain choline acetyltransferase (ChAT) were obtained. Immunoblot analysis indicated that both antibodies revealed two adjacent protein bands on a sodium dodecyl sulfate-polyacrylamide gel electrophoresis. ChAT was demonstrated immunohistochemically by one of the antibodies in the motoneurons in the monkey brainstem and spinal cord. This antibody also revealed ChAT-positive terminal-like structures in the neuropils of lamina IX of the cervical spinal cord and interpeduncular nucleus.     

Morphological study of parafollicular cells and the parathyroid gland in the house shrew (Suncus murinus).

A morphological study of parafollicular cells in the thyroid gland and parathyroid gland of the house shrew (Suncus murinus) was made. The results indicated that (1) there were two pairs of parathyroid glands which were located in the upper part of the house shrew thyroid gland, (2) the volumes of the house shrew parathyroid glands ranged from 0.014 to 0.079 mm3, (3) the number of parafollicular cells along the follicles was largest in the upper part of the thyroid lobe, while no parafollicular cells were present in the isthmus, and (4) there were about 20 parafollicular cells per 100 follicular cells and 1.519 parafollicular cells per follicle. The number of parafollicular cells per 100 follicular cells was thus about 5 times larger than that in rats and the number of parafollicular cells per follicle about 2.5 times larger than that in rats.     

Expression profiling of cytokines and related genes in regenerating skeletal muscle after cardiotoxin injection: a role for osteopontin

To examine the roles of cytokines in muscle regeneration, we injected cardiotoxin into mouse tibialis anterior muscle and examined the expression profiles of cytokines and related genes in the regeneration process. Expression of 40, 64, and 7 genes among 522 genes spotted on a cytokine expression array were increased more than fivefold at 48 hours, 96 hours, and 7 days after toxin injection, respectively, when compared with those of the control muscle. Especially the levels of mRNA for chemokines and chemokine receptors, many of which are potent regulators of macrophages, were highly elevated 48 hours after injury. The expression of osteopontin (OPN), a versatile regulator of inflammation and tissue repair, was up-regulated more than 118-fold in regenerating muscle at 48 hours after injury. Northern blotting confirmed that the expression of OPN was highest at 48 hours after cardiotoxin injection and declined sharply thereafter. Immunohistochemistry showed that OPN was detected both in the cytoplasm of macrophages and in necrotic muscle infiltrated with macrophages. Our studies suggest OPN may serve as an adhesion molecule that promotes macrophage binding to necrotic fibers and may be an important mediator in the early phase of muscle regeneration.     

Comparison of various bone marrow fractions in the ability to participate in vascular remodeling after mechanical injury

In contrast to conventional assumption, recent reports propose the possibility that hematopoietic stem cells (HSCs) may have broader potential to differentiate into various cell types. Here, we tested the pluripotency of HSCs by comparing vascular lesions induced by mechanical injury after bone marrow reconstitution with total bone marrow (TBM) cells, c-Kit+ Sca-1+ Lin- (KSL) cells, or a single HSC cell (Tip-SP CD34-KSL cell, CD34- c-Kit+ Sca-1+ Lin- cell with the strongest dye-efflux activity) harboring green fluorescent protein (GFP). The lesions contained a significant number of GFP-positive cells in the TBM and KSL groups, whereas GFP-positive cells were rarely detected in the HSC group. These results suggest that transdifferentiation of a highly purified HSC seems to be a rare event, if it occurs at all, whereas bone marrow cells including the KSL fraction can give rise to vascular cells that substantially contribute to repair or lesion formation after mechanical injury.     

Peliosis Hepatis in a Child with X-Linked Myotubular Myopathy Treated with Living-Donor Liver Transplant: A Case Report

BackgroundMyotubular myopathy is a rare disease sometimes accompanied by peliosis hepatis, a leading cause of fatal liver hemorrhage.Case ReportWe present a case of a 2-year-old boy with myotubular myopathy who developed liver hemorrhage because of peliosis hepatis and was successfully treated with living-donor liver transplant. The patient initially presented with fever, anemia, and liver dysfunction. A computed tomographic scan revealed hemorrhages in the liver, and the patient underwent hepatic artery embolization twice. After the second embolization, multiple peliosis hepatis cavities appeared in the left lobe of the liver that had increased in size. Therefore, the patient underwent ABO-incompatible living-donor liver transplant using a lateral segment graft from his father. The patient developed severe septic shock with an unknown focus on postoperative day 18, which resolved with antibiotic therapy. On postoperative day 62, he was discharged. Fourteen months after undergoing living-donor liver transplant, the patient showed no recurrence of peliosis hepatis.ConclusionsAlthough the long-term prognosis of peliosis hepatis due to myotubular myopathy after living-donor liver transplant remains unclear, liver transplant may be a curative treatment for patients with myotubular myopathy who have uncontrollable peliosis hepatis.     

Influences of intra-aortic balloon pumping on superior mesenteric flow dynamics

The influences of intra-aortic balloon pumping (IABP) on arterial flow of the superior mesenteric artery were assessed by Doppler echocardiography. The subjects were 13 patients postoperatively, requiring IABP support to control low cardiac output state, in which distal aspects of balloons were distal to the superior mesenteric artery. Superior mesenteric flow velocity integral in systole (IntS) and that in diastole (IntD) were measured from superior mesenteric flow pattern, and the sum of IntS and IntD (IntS + IntD) was calculated ON and OFF balloon pumping (IABP ON-OFF test). The same parameters were obtained with balloon inflating on every other beat (IABP 1:2 test); the cardic cycle with balloon assist was defined as "1:2 ON", and that without balloon assist was defined as "1:2 OFF". 1) IABP ON-OFF test. IABP increased IntS from 7.07 +/- 2.56 cm to 9.20 +/- 3.19 cm (p less than 0.05), IntD from 3.00 +/- 1.18 cm to 3.62 +/- 1.40 cm (p less than 0.05), and IntS + IntD from 10.07 +/- 3.48 cm to 12.82 +/- 4.04 cm. Cardiac output increased from 3.89 +/- 1.34 l/min to 4.24 +/- 1.64 l/min with IABP support. The increments in IntS, IntD and IntS + IntD with IABP are attributed, to a large extent, to an increase in cardiac output. 2) IABP 1:2 test. Without balloon inflation, IntS increased (1:2 ON; 7.16 +/- 2.91 cm, 1:2 OFF; 8.41 +/- 3.30 cm, p less than 0.05), and IntD decreased (1:2 ON; 3.51 +/- 1.60 cm, 1:2 OFF; 2.33 +/- 1.25 cm, p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)