Imbalance between production and scavenging of hydroxyl radicals in patients maintained on hemodialysis

Background Reactive oxygen species are as being related to the pathophysiology of endstage renal disease (ESRD). We measured the plasma hydroxyl radical (·OH)-producing ability and ·OH-scavenging activity in patients with ESRD to clarify the pathophysiological states involved. Methods We used electron spin resonance to measure plasma N-t-butyl-α-phenylnitron radical spin adduct (pPBN rsa) as ·OH-producing ability and plasma 3,3,5,5-tetramethyl-1-pyrroline-N-oxide radical spin adduct (pM4PO rsa) as ·OH-scavenging activity. Oxidative injuries were evaluated by determining oxidised low-density lipoprotein (Ox-LDL). Results The pPBN rsa of the ESRD patients was lower than that of the controls (1.83 vs 2.94 μmol/g protein). The pM4PO rsa of the ESRD patients was higher than that of the controls (3.85 vs 3.15 mmol l-ascorbic acid 2-[3,4-dihydro-2,5,7,8-tetramethyl-2-(4,8,12-trimethyltridecyl)-2H-1-benzopyran-6-yl hydrogen phosphate] potassium salt (EPC-K₁)/g protein). The pPBN rsa and pM4PO rsa were correlated, both in the ESRD patients and in the controls (r = 0.47 and r = 0.53). Ox-LDL was correlated with hemodialysis (HD) duration (r = 0.49) and was negatively correlated with pPBN rsa (r = −0.54), which indicates that oxidative stress was increased as HD therapy was prolonged and suppressed pPBN rsa. Conclusions There was an imbalance between ·OH-producing ability and ·OH-scavenging activity, in the ESRD patients, and this may be responsible for compromising the health of ESRD patients.     

Prenatal diagnosis of I-cell disease by measuring altered α-Mannosidase activity in amniotic fluid

Prenatal diagnoses of I-cell disease were carried out by examination of the amniotic fluid and cultured amniotic cells in four cases of high-risk pregnancy in three different families. Three of the four fetuses were diagnosed as having I-cell disease, on the basis of observation of abnormally increased activity of lysosomal acid hydrolases in the amniotic fluid and their decreased activity in cultured amniotic cells, thus leading to therapeutic abortion. α-Mannosidase in the amniotic fluid of the fetuses with I-cell disease exhibited significant alterations in pH profile,K _m value, thermal stability and isoelectric focusing pattern, compared with the enzyme in normal controls. The results indicate that prenatal diagnosis of I-cell disease may be accomplished by demonstration of altered enzymological characteristics of α-mannosidase in the supernatant of amniotic fluid. The significance of the alteration of α-mannosidase in the amniotic fluid is not known.     

Giant basilar aneurysm--a case report (author's transl)]

A case of a giant aneurysm arising from the basilar artery is presented. The patient a 41--year-old female, had recent memory disturbance, miscalculations and hemiparesis on admission. Vertebral angiograms displayed a giant aneurysm measuring approximately 66 mm in diameter arising from the basilar artery, a tortuous vertebral artery, an extremely dilated basilar artery and an agenesis of the left carotid artery. Due to the enormous size of the aneurysm and its location in the deep intracranial portion, radical surgery was abandoned. Six months following her discharge, she died. At autopsy, the giant aneurysm arising from the top of the basilar artery was found, existing just beneath the third ventricle and extending to the left basal ganglia. The lateral ventricle, the third ventricle and the Sylvian fissure were filled by hematoma resulting from the rupture of the aneurysm. It is possible that this giant aneurysm could be considered a vascular anomaly, and hence be distinguished from saccular aneurysms.     

Early onset distal muscular dystrophy with normal dysferlin expression

A 7-year-old boy, who was noted to be a slow runner at the age of 2 years, had progressive muscle weakness and atrophy, preferentially affecting distal muscles. At 3 years of age, he had scoliosis and difficulty in standing on tip-toe. Serum creatine kinase was 1074 IU/l. Muscle CT scan showed low-density areas in the lower legs and upper arms, but predominantly in the gastrocnemius and soleus muscles. Biopsy of the biceps brachii muscle showed moderate dystrophic changes with normal dysferlin expression on immunohistochemical and western blot analyses. Although muscle involvement mimicked that seen in Miyoshi myopathy (MM), the very early onset of the disease and scoliosis were quite unusual for MM. We, therefore, made the diagnosis of early onset dysferlin-positive distal muscular dystrophy, probably a new type of distal muscular dystrophy.     

Chiari I malformation associated with ventral compression and instability: one-stage posterior decompression and fusion with a new instrumentation technique

OBJECTIVE AND IMPORTANCE: We describe an alternative surgical technique for treatment of Chiari I malformation associated with ventral compression and instability of the region. An expansive suboccipital cranioplasty and a rigid occipitocervical fixation are performed in one stage. METHODS: The occipitocervical fixation is performed by use of metal rods fixed on the cranial side by screws inserted into the diploic layer of occipital bone and on the caudal side by screws inserted into the pedicle of the axis or in a transarticular fashion into the lateral masses of axis and atlas vertebra. A large piece of autologous bone is placed in the region between the rostral edge of cranial decompression and the axis, with the aim of achieving both expansive suboccipital cranioplasty and occipitocervical fusion. RESULTS: We performed this procedure in two patients with Chiari I malformation associated with basilar invagination and occipitalization of the atlas. Postoperatively, decompression of the brainstem and restoration of normal cerebrospinal fluid flow at the craniovertebral junction were confirmed radiologically, and the patients were relieved of their symptoms. At 1 and 3 years of follow-up, respectively, solid bone fusion was observed between the occipital bone and axis in both patients. CONCLUSION: Simultaneous posterior decompression and occipitocervical fixation with an alternative instrumentation technique is discussed. The procedure can be performed regardless of the size of suboccipital craniectomy. Screw insertion into the diploic layer of the occipital bone has not been described previously.     

New qualitative detection methods of genetically modified potatoes

In Japan, 8 lines of genetically modified (GM) potato (2 lines of NewLeaf potato; NL, 3 lines of NewLeaf Plus potato; NLP, and 3 lines of NewLeaf Y potato; NLY) have already been authorized as safe for use in foods and feeds. We have developed polymerase chain reaction (PCR) methods for the qualitative detection of the GM potatoes for the screening and the identification of NL, NLP and NLY. The gene encoding uridine diphosphate (UDP)-glucose pyrophosphorylase (UGPase) was used as a taxon specific gene. We designed the primer pair to detect the cryIIIA genes as a screening method for GM potatoes because the gene should be inserted in all 8 lines of the GM potatoes. For identification of NL, NLP and NLY, we further designed three specific primer pairs for the different recombinant DNAs (r-DNA) specifically introduced into NL, NLP, or NLY. In addition, to identify the 3 lines of NLY that have been introduced with the same r-DNA, the three line-specific primer pairs for the border sequence between the r-DNA and genomic DNA of NLY 3 lines were designed. Six lines of GM potato used as the test material were specifically identified using the each primer pair under the same PCR condition. The detection limits of all the GM potatoes should be approximately 0.1%. Furthermore, the specificity and reproducibility of the methods were confirmed in a six-laboratory collaborative study.     

Generation of a transgenic animal model of hyperthyroid Graves' disease

Graves' disease (GD) is an organ-specific autoimmune disease characterized by hyperthyroidism. Agonistic anti-thyrotropin receptor antibodies (thyroid-stimulating antibodies, TSAb), which mimic the thyrotropin (TSH) action, are thought to cause GD. The precise immunological mechanism of TSAb production, however, remains elusive. Previous immunization approaches using TSH receptor led to transient hyperthyroidism, but did not seem sufficient for comprehensive understanding of the development of autoimmune responses. To create GD-related autoimmunity in mice, we here generated TSAb-transgenic mice in which a patient-derived TSAb is expressed in B cells. Expression of the human TSAb in mice resulted in various manifestations of hyperthyroidism including increased free thyroxine levels with concomitantly decreased TSH levels, increased thyroid uptake of technetium pertechnetate, hyperthermia and thyroid hyperplasia. We found a correlation between the serum levels of human TSAb immunoglobulin and free thyroxine. In addition, conventional B cells expressing the TSAb were partially deleted in the periphery while B1 cells expressing the TSAb persisted and accumulated in the peritoneal cavity, a finding consistent with previous demonstrations that the maintenance of B1 cells plays an important role in the development of autoimmune diseases. Thus, our transgenic mouse may provide a novel and useful animal model for elucidating the pathogenesis and pathophysiology of GD.     

The role of ficolins in innate immunity

Ficolins are a group of proteins containing both a collagen-like domain and a fibrinogen-like domain and are found in varying tissues. Ficolins present in sera have a lectin activity toward N-acetylglucosamine through their fibrinogen-like domains. The domain organizations between ficolins and mannose-binding lectin (MBL) are very similar in that both consist of a collagen-like domain and a carbohydrate-binding domain, although their carbohydrate-binding moieties are different. MBL acts as an opsonin and activates complement in association with MBL-associated serine proteases (MASPs) and sMAP, a truncated protein of MASP-2, via the lectin pathway. Like MBL, two types of human serum ficolins, L-ficolin/P35 and H-ficolin, are associated with MASPs and sMAP, and activate the lectin pathway. In addition, L-ficolin/P35 acts as an opsonin. These findings indicate that serum ficolins play an important a role in innate immunity in a similar manner to MBL.