Production and characterization of monoclonal antibodies against amino-terminus of human alpha-atrial natriuretic polypeptide

Monoclonal antibodies directed against human, alpha-atrial natriuretic polypeptide (alpha-ANP; Human, 1-28) were obtained by somatic cell fusion between P3-X63-Ag8.653 myeloma cells and spleen cells from a BALB/c mouse immunized with human, alpha-ANP selectively coupled to keyhole limpet hemocyanin. From the analysis of polyclonal sera with respect to determinant specificity before the fusion, the strategy was primarily used to pick up monoclonal antibody specific for the N-terminal residues of human, alpha-ANP. Screening of antibodies in the hybridoma culture supernatants were performed by binding to iodinated synthetic human, alpha-ANP. Two stable clones producing anti-human, alpha-ANP antibodies, designated 13A1 and 10B1, were obtained by the limiting dilution technique. The ability of ANP(Rat, 1-28) to inhibit binding of 125I-human, alpha-ANP to these antibodies was almost equipotent to ANP(human, 1-28). However, ANP fragments (Human, 7-28) and (18-28) did not compete the binding completely. These results suggest that both 13A1 and 10B1 monoclonal antibodies can specifically recognized N-terminus of human, alpha-ANP, and may be a useful tool to investigate receptor binding of human, alpha-ANP by the antagonizing effect.     

Superoxide dismutases of muscle in mitochondrial encephalomyopathies

Lmmunohistochemical analyses were made of the superoxide dismutases (Mn-SOD and CuiZn-SOD) in biopsied muscles from 7 patients with mitochondrial encephalomyopathies that included mitochondrial encephalomyopathy, lactic acidosis and strokelike episodes (MELAS), and chronic progressive external ophthalmoplegia (CPEO). Mn-SOD mainly was present in the subsarcolemmal region, but it also was found in a coarsely granular, reticular, or diffuse pattern of staining within the muscle fibers. These Mn-SOD-positive fibers corresponded almost completely to the ragged-red fibers. The immunoreaction for CuiZn-SOD was weakly positive in some of the muscle fibers positive for Mn-SOD. In CPEO, Mn-SOD-positive fibers predominantly showed decreased cytochrome c oxidase (COX) activity. In MELAS, Mn-SOD-positive fibers tended to be stained deeply for COX although a few were COX-negative. These findings suggest that Mn-SOD-positive fibers can be used to make a differential diagnosis between CPEO and MELAS and that in mitochondrial encephalomyopathies Mn-SOD in the raggedred fibers may protect against oxidative stress. © John Wiley & Sons, Inc.     

MR imaging of dural arteriovenous malformations with ocular signs

Summary Four patients with dural arteriovenous malformation (AVMs) draining into the cavernous sinus, who presented ophthalmic manifestations, were studied by magnetic resonance (MR) imaging. In all patients signal decrease in the involved cavernous sinus was demonstrated in coronal spinecho (SE) imaging. It is attributable to rapid venous flow in the sinus, and this high velocity signal loss is a fairly pathognomonic finding in this condition. We stress the validity of MR imaging in the primary diagnosis of dural AVMs with ophthalmic symptoms.     

The effects of an intracellular calcium antagonist HA 1077 on delayed cerebral vasospasm in dogs

Summary The effectiveness of calcium antagonists on a chronic cerebral vasospasm after an SAH is still under debate. Calcium channel blockers such as nimodipine, nefedipine etc. can dilate spastic arteries by intrathecal administration, but not by systemic (iv or po) use. HA 1077 is a novel and potent calcium antagonist vasodilator which is considered to act by employing different mechanisms from the usual calcium channel blockers since it inhibits 1. calcium ionophore A 23187 induced contraction in arterial strips and 2. phenylephrine induced contraction in calcium free media, suggesting that its site of action is in the intracellular space. HA 1077 is water soluble and relatively stable in light.In the present study, the efficacy of HA 1077 was evaluated on dogs by using the spiral arterial strips in vitro and by angiography in vivo. In the arterial strips from the control dogs, a 50% relaxation of KCl (15 mM) induced contraction was obtained by a 10^–6 M HA 1077 for the intracranial basilar and middle cerebral arteries, while a 10^–5 M was needed to obtain the same effect for the extracranial common carotid and vertebral arteries, indicating that HA 1077 is more effective for the intracranial arteries. A vasospasm was produced by the two haemorrhage model of Varsoset al. The average angiographic diameter of the basilar artery was reduced to 60% of the control on SAH day 7. Intravenous infusion of HA 1077 (0.5–3 mg/kg/30 min) significantly dilated the spastic basilar artery (up to 20–30%), for over 2 hours. A fall in the systemic BP remained less than 20% during this time. Such spasmolytic effects by intravenous administration could not have been obtained with the usual calcium channel blockers. HA 1077 may be suitable for the treatment of a vasospasm in humans as well.     

The effect of RO15-1788 on cardiovascular depression caused by fentanyl and diazepam

Cardiovascular depression occuring when diazepam is combined with fentanyl has been investigated using the benzodiazepine antagonist RO15-1788 in the dog.After the initial administration of fentanyl (40mcg/kg), the mean arterial pressure (MAP) decreased to 89% of its control value. Following the administration of diazepam (1.2mg/kg), the MAP and the total peripheral resistance (TPR) decreased significantly, to 75% and 83% of their control values respectively. After the administration of RO15-1788 (0.4mg/kg), the MAP increased significantly to 90% and the TPR to 102% of their control values and, lastly, the administration of naloxone (40mcg/kg) increased the MAP to 108% of its control value. No relationship was found between the changes in the catecholamines and the changes in the MAP after the administration of fentanyl, diazepam, and RO15-1788.The mechanism of circulatory depression when diazepam was used with fentanyl is interpreted as being a peripheral vasodilatory effect of diazepam acting by way of the benzodiazepine receptors since RO15-1788 was found to antagonize this effect.(Sone T, Kato T, Tsukahara I et al.: The effect of RO15-1788 on cardiovascular depression caused by fentanyl and diazepam. J Anesth 2: 69–76, 1988)     

Correlation between vibration-induced white finger and symptoms of upper and lower extremities in vibration syndrome

Summary The correlation was investigated between the frequency of attacks of vibration-induced white finger (VWF) and numbness or coldness of the fingers and legs in patients with vibration syndrome. Some 1687 patients with vibration syndrome were examined and of these 342 chain-saw operators and 277 rock-drill operators had no disease other than vibration snydrome. Then subjects were matched by age and period of treatment within three years. In the last analysis, 20 in the VWF almost everyday group or in the never group, and 40 in the occasionally group were selected from the chain-saw operators, and from the rock-drill operators 32 in the VWF everyday or the never group and 64 in the occasionally group. The present study showed that, with the frequency of VWF attacks, patients had a higher prevalence of coldness not only in the fingers but also in the legs. These findings suggest a correlation between the severity of circulatory disturbances of the upper extremities and that of the lower ones in patients with vibration syndrome. Further studies on circulatory disturbances in the leg are required.     

Evidence for a role of basal ganglia in the regulation of rapid eye movement sleep by electrical and chemical stimulation for the pedunculopontine tegmental nucleus and the substantia nigra pars reticulata in decerebrate cats

The present study was to determine how afferents from the substantia nigra pars reticulata (SNr) of the basal ganglia to the pedunculopontine tegmental nucleus (PPN) in the brainstem could contribute to the control of behavioral states. We used anesthetized and acutely decerebrated cats (n=22). Repetitive electrical stimulation (10-100 Hz, 20-50 microA, for 4-20 s) to the ventrolateral part of the PPN produced rapid eye movement (REM) associated with a suppression of postural muscle tone (REM with atonia). Although repetitive electrical stimuli (10-200 Hz, 10-60 microA, for 5-20 s) delivered to the dorsolateral part of the SNr did not evoke eye movements or muscular tonus in baseline conditions, it altered the PPN-induced REM with atonia. The following three types of effects were induced: (1) attenuation of the REM with atonia; (2) attenuation of muscular atonia without changes in REM (REM without atonia); and (3) attenuation of only REM. The optimal stimulus sites for these effects were intermingled within the lateral part of the SNr. The PPN-induced REM with atonia was abolished by an injection into the PPN of muscimol (1-15 mM, 0.1-0.25 microl), a GABAA receptor agonist, but not altered by an injection of baclofen (1-10 mM, 0.1-0.25 microl), a GABAB receptor agonist. Moreover, an injection of bicuculline (1-15 mM, 0.1-0.25 microl), a GABAA receptor antagonist, into the PPN, resulted in REM with atonia. On the other hand, an injection of muscimol into the dorsolateral part of the SNr (1-15 mM, 0.1-0.25 microl) induced REM with atonia, which was in turn eliminated by a further injection of muscimol into the PPN (5-10 mM, 0.2-0.25 microl). These results suggest that a GABAergic projection from the SNr to the PPN could be involved in the control of REM with atonia, signs which indicate REM sleep. An excessive GABAergic output from the basal ganglia to the PPN in parkinsonian patients may induce sleep disturbances, including a reduction of REM sleep periods and REM sleep behavioral disorders (REM without atonia).     

Evaluation of dengue IgM detection tests using sera from patients with autoimmune diseases

Three commercial dengue IgM test kits and 'in-house' IgM-capture enzyme-linked immunosorbent assay (ELISA) were examined for false positive reactions, using 49 serum samples from patients with autoimmune diseases. All the samples were found to be negative by the 'in-house' IgM-capture ELISA. Five samples were determined to be positive by the immunochromatographic test and three of the five samples were also found positive by one commercial IgM-capture ELISA kit. These results suggest that a possibility of false positive reaction should be considered when serum samples from autoimmune disease patients are tested for dengue IgM by some commercial dengue IgM test kits.     

Pulmonary angiitis with atypical lymphoreticular infiltrates in Wiskott-Aldrich syndrome: possible relationship of lymphomatoid granulomatosis and EBV infection

We describe a 12-year-old boy with Wiskott-Aldrich syndrome who developed a pulmonary vasculitis associated with lymphoreticular proliferation, consistent with the histological and clinical diagnosis of lymphomatoid granulomatosis. The lesions were responsive to cyclophosphamide and steroids. The patient has had severely depressed immune function and was shown to have abnormal Epstein-Barr virus (EBV)-specific cellular and humoral immune responses. Lymph nodes obtained at autopsy were positive for EBV genome. In this patient, reactivated EBV infection resulting from impaired immune surveillance of the virus may have been responsible for the development of this paraneoplastic disorder.