A Sensitive and Simple Determination of Human α-fetoprotein by Enzyme-electroimmunodiffusion

A simple and sensitive determination method of α-fetoprotein was developed by introducing α-fetoprotein horseradish peroxidase conjugate into electroimmunodiffusion.
The precipitin line formed was directly visualized by staining for peroxidase activities. The sensitivity was 32 ng/ml. Electrophoretic activities of AFP were analyzed by two dimensional electrophoresis applying the present method to the second electrophoresis.
AFP from normal adults and several patients was confirmed to migrate as an α-globulin as AFP of hepatoma, yolk sac tumor and fetuses did.
AFP-like activities were detected in β-γ-globulin region in some samples but these were not due to AFP-anti-AFP interactions.     

Clinical profiles of autopsy-confirmed dementia with Lewy bodies at institutionalization: Comparison with Alzheimer's disease

Background: It has been reported that Alzheimer’s disease (AD) patients with Lewy pathology have a shorter time from a given baseline to institutionalization than those with AD alone. Taking the clinical distinction between dementia with Lewy bodies (DLB) and AD into consideration, the previous findings may indicate the possibility that the clinical characteristics of DLB patients have an influence on early institutionalization. This study was carried out to clarify whether there are any differences in the symptoms that required institutionalization between patients with DLB and those with AD. Methods: Hospital records and standardized data forms completed at admission to a residential care facility were reviewed to assess the profiles in all cases with autopsy‐confirmed diagnoses for correct differential diagnosis. We examined functional, cognitive and symptomatic conditions at admission to a residential care facility of 18 DLB and 35 AD patients whose diagnoses were confirmed by autopsy. The examinations were conducted using the Gottfries‐Bråne‐Steen (GBS) scale and cognitive tests, and the results were compared between the two groups of patients. Results: Hallucinations, impaired wakefulness, disturbance of ADL and emotional disturbance, common clinical features compatible with DLB, were more frequently observed in DLB patients than in AD patients (P < 0.05). Moreover, DLB patients had higher scores on cognitive tests than did AD patients at admission to a residential care facility (P < 0.05). Conclusion: The distinctive clinical features at admission to a residential care facility may indicate that the reasons for the necessity of institutionalization are different between DLB and AD, and that the interventions specific to DLB patients and their families would be necessary to prevent or postpone institutionalization.     

Stable Production of Recombinant Human Sperm Immobilizing Antibody Using cDNA Expression Vectors

PROBLEM: Sperm immobilizing antibodies present in sterile women may be one of the principal causes of immunological infertility. We already established cell lines that secrete recombinant human IgG sperm immobilizing antibody using class-switched (from IgM to IgG) genomic immunoglobulin genes. However, these transfectants produced a small quantity of antibody and required continuous use of a medium with selective reagents. We have now constructed cell lines that stably secrete the antibody in large quantities using immunoglobulin cDNAs and cDNA expression vectors. METHOD: The immunoglobulin heavy chain cDNA was cloned from transfectants that secrete the class-switched human IgG sperm immobilizing antibody. The light chain cDNA, which had already been cloned, and the heavy chain cDNA were inserted into the modified bovine papilloma virus-based cDNA expression vectors BCMGSNeo and BCMGSHyg, respectively. These constructs were sequentially transfected into a mouse myeloma cell line by electroporation. RESULTS: The established transfectants produced recombinant antibody that retained human sperm immobilizing activity in nonselective medium for at least 30 days. Moreover, the production of the antibody was increased three times over that of the previous cell lines. CONCLUSION: We have established an unique method that improves the production of sperm immobilizing antibody stably and in large quantities.     

Characterization of fertilization-blocking monoclonal antibody 1G12 with human sperm-immobilizing activity

A mouse hybridoma (1G12) producing sperm-immobilizing MoAb to human sperm was established and characterized in order to study the antigens relevant to sperm immobilization by antibodies. MoAb 1G12 had strong sperm-immobilizing and agglutinating activities and also showed a fertilization-blocking activity on in vitro fertilization tests. The antibody absorption experiments showed that MoAb 1G12 reacted not only to ejaculated sperm but also human seminal plasma, suggesting that the corresponding antigen might be a sperm coating antigen. The MoAb also reacted with peripheral blood lymphocytes. In histochemical studies, the epithelia of corpus epididymis were most strongly stained. Ejaculated sperm were stained with a granular pattern for their entire surface by immunofluorescence. MoAb 1G12 recognized polymorphic glycoproteins of 15–25 kD in the ejaculated sperm extract in Western blot analysis. After deglycosilation of the sperm extract, only a single staining band of under 15 kD was detected by MoAb 1G12. This suggests that the antigen epitope recognized by MoAb 1G12 might be a peptide of the core portion of the glycoprotein. MoAb 1G12 might be a useful tool for studying the mechanism of egg–sperm interaction, and also be applied to identifying the corresponding antigen by using gene technology.     

Moebius syndrome associated with pituitary dwarfism and hypoplastic optic disc

A 17 year old male patient with Moebius syndrome with pituitary dwarfism and unilateral hypoplastic optic disc is presented. Although there have been several reports of an association of Moebius syndrome and pituitary dysfunction, growth hormone deficiency has not been reported previously. These associations may give some insight into the pathogenesis of Moebius syndrome.     

Increased incidence of dementia with Lewy bodies in patients carrying the ɛ4-allele of apolipoprotein E

Background: The apolipoprotein ∈4 (APOE4) allele is a risk factor for Alzheimer's disease, but it remains undetermined whether this allele is related to the pathological development of neurofibrillary tangles (NFT) and the formation of Lewy bodies. Methods: In the present study, we examined the relationship between these changes and the APOE4 allele in 255 consecutive neuropathologically diagnosed cases. APOE genotyping was carried out by the polymerase chain reaction–restriction fragment length polymorphism method. Results: Nearly all our cases of dementia with Lewy bodies (DLB) showed the common form, having numerous senile plaques in the cerebral cortex and NFT in the parahippocampal and hippocampal regions and were also associated with the APOE4 allele. Limbic neurofibrillary tangle dementia (LNTD), characterized by the presence of NFT in limbic areas as well as the absence of senile plaques, did not appear to be associated with the APOE4 allele. Conclusions: The APOE4 allele is a risk factor for DLB as well as Alzheimer's disease and cerebral amyloid angiopathy, but not for LNTD.     

Direct Production of the Fab Fragment Derived From the Sperm Immobilizing Antibody Using Polymerase Chain Reaction and cDNA Expression Vectors

PROBLEM : Sperm immobilizing antibodies cause infertility mainly through complement dependent sperm immobilization. To analyze any effect of sperm immobilizing antibody on fertilization, we had already established cell lines that secrete IgM monoclonal antibody (MAb H6-3C4) and IgG monoclonal antibody (MAb EnBCMGS). The latter was a class-switched recombinant IgG antibody that shares the same variable region as MAb H6-3C4. The biological effects of the IgG antibody were also reported previously to eliminate sperm immobilizing or sperm agglutinating activities. However, the method of chemical digestion of IgG had some disadvantage to prepare the purified Fab fragment stably and in large quantities. This time we report a unique method to obtain the recombinant Fab fragments (Fab EnBCMGS) using polymerase chain reaction (PCR) and cDNA expression vectors. METHOD : Two kinds of PCR primers were designed to make a truncated heavy chain (Fd) gene of MAb EnBCMGS. The amplified Fd gene and light chain gene were ligated into cDNA expression vectors and then transfected into mammalian cells. RESULTS : Expression of the Fd gene and light chain gene were confirmed by Northern blotting. Secretion of the recombinant Fab fragment from mammalian cells was also confirmed by Western blotting. The Fab fragment showed biological activity as is expected by FACS analysis. CONCLUSION : This method enables the stable production of genuine Fab fragments of IgG in mammalian cells without any chemical treatment that may be time consuming and affect the quality of the Fab fragments.     

TUMOR OF GLOMUS JUGULARE REPORT OF A CASE

A typical case of glomus jugulare tumor was presented. This 42-year-old man suffering from a left-sided Impaired hearing progressive over a Ave year period was found to have a small hemorrhagic polypoid tumor in the left external auditory canal. After repeated surgical treatments the tumor removed was light and electron microscopically diagnosed as glomus jugulare tumor which corresponded to the 13th case in this country. The Implication that this tumor may constitute a distinctive group of tumor among the neuroendocrine tumors from other sites was discussed from the facts of the presence of a large number of rather small argyrophilic granules of secretory type in the cytoplasm of tumor cells including the present case and of the appearance of increasing cases with functioning glomus jugulare tumor secreting catecholamine in the recent medical literature.