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.     

Gastritis Cystica Profunda Histopathological Study of Nine Cases

Nine cases of gastritis cystica profunda (diffuse submucosalproliferation of cystic glands) were studied. Three of the patientshad coexisting carcinoma, four had peptic ulcers, and the othertwo had atrophic gastritis. All of them were middle aged orelderly, except for one postoperative young male. Morphologicalcharacteristics of the submucosal cystic nodules are described,and their pathogenesis is discussed in relation to the intestinalmetaplasia of the gastric mucosa.     

Controlled Study of Therapeutic Granulocyte Transfusions in Granulocytopenic Patients with Severe Infections

The effectiveness of therapeutic granulocyte transfusions wasstudied in a controlled trial involving 75 granulocytopenicpatients with severe infections. Patients who had granulocytecounts of less than 200/mm³ and no response to antibiotic therapywere assigned to receive antibiotic therapy alone or granulocytetransfusions plus antibiotic therapy. Granulocytes were collectedby filtration leukapheresis (FL), intermittent flow centrifugeleukapheresis (IFCL) or continuous flow centrifuge leukapheresis(CFCL). Therapeutic effects of granulocyte transfusions wereevaluated on the basis of 20-day survival after entry into thestudy. The patients were divided into three groups: 1) 22 patientsreceived antibiotic therapy alone for 29 infectious episodes(control group); 2) 27 patients received 131 transfusions ofgranulocytes collected by FL for 31 infectious episodes (FLgroup); 3) 26 patients received 139 transfusions of granulocytescollected by IFCL and CFCL for 27 infectious episodes (IFCL& CFCL group). The overall survival of the transfused patientswas not significantly different from that of the untransfusedpatients. Similarly, there was no significant difference insurvival between the transfused and untransfused patients whenthe data concerning septicemia were analyzed. When patientsshowed bone-marrow recovery, which was indicated by recoveryof granulocytes to 500/mm³ or more during the study, 20-daysurvival was observed in 13 of 14 untransfused patients andin all of 26 transfused patients. In contrast, the survivalrate for the patients without bone-marrow recovery was 13% (2/15)in the control group, 39% (7/18) in the FL group and 57% (8/14)in the IFCL & CFCL group respectively. Thus, the survivalrate was significantly higher for the transfused than for theuntransfused patients. These observations suggest that therapeutic granulocyte transfusionsmay be of limited value in improving the outcome of severe infectionsin persistently granulocytopenic patients. Since bone-marrowrecovery is a critical factor for the prognosis of severe infections,therapeutic granulocyte transfusions do not provide any beneficialeffects in granulocytopenic patients whose bone-marrow functionwill recover.     

Increased Mast Cell Density in Peritoneal Endometriosis Compared with Eutopic Endometrium with Endometriosis

PROBLEM: Factors regulating fibrosis in peritoneal endometriosis are poorly understood. We hypothesized that mast cell proteases may play a role in the process of fibrosis in peritoneal endometriosis. As the first step in examining the possible contribution of mast cells to fibrosis, we investigated their distribution in peritoneal endometriosis lesions according to their macroscopic appearance. METHOD OF STUDY: Mast cells were identified by immunohistochemistry using monoclonal antibody against mast cell tryptase on formalin-fixed, paraffin-embedded sections. Mast cell density (mean mast cell count per 0.13 mm² of stroma) was measured using a computerized image analysis system. RESULTS: Mast cell density was significantly increased in black peritoneal lesions compared with red peritoneal lesions in endometriosis. CONCLUSIONS: Mast cells may be involved in the pathogenesis of peritoneal endometriosis.     

Analysis of the mandibular relationship of TMJ dysfunction patients using the Mandibular Kinesiograph

Analyses of the relationship between centric relation and centric occlusion were performed using the Mandibular Kinesiograph to show the abnormality of mandibular positions of the TMJ dysfunction patients. The distance between centric relation and centric occlusion in antero-posterior, left, and linear direction showed significant differences between the normal subjects and the TMJ dysfunction patients. In the TMJ dysfunction patients, the number of the subjects who showed larger deviation between centric relation and centric occlusion increased remarkably when compared with normal subjects. It is suggested that the slide between centric relation and centric occlusion can be one of the causes of the TMJ dysfunction.     

Synthesis and solubility properties of peptide fragments of human hemoglobin α-chain (123-136)

The solubility prediction method for protected peptides was successfully applied to relatively small peptide fragments of human hemoglobin α-chain (123-136) which contained various polar amino acid residues such as Asp(OBzl), Glu(OBzl), Lys(Z), Ser(Bzl), and Thr(Bzl). As reported previously for hydrophobic peptides and human proinsulin C-peptide fragments, solubility data indicated that the insolubility of protected peptides having a <P_C > value below 0.90 appeared to begin at the octa- or nonapeptide sequence level and that β-sheet structure played an important role in the insolubility of peptides. When a peptide has a β-sheet structure in the solid state, we can clearly determine the critical chain length for peptide insolubility, the solubility dependence on solvent properties, and the solubility independence of amino acid compositions of peptides.     

Comparison of Behavioural Effects of Repeated Treatment with Methamphetamine plus Scopolamine and Methamphetamine Alone on Behavioural Sensitization and Conditioned Response

We investigated how repeated treatments with methamphetamine (4.0mgkg^?1, i.p.) plus scopolamine (0.5mgkg^?1, i.p.) and methamphetamine alone effected behavioural sensitization and conditioned response in rats. Repeated methamphetamine plus scopolamine treatment induced a more progressive and enduring enhancement of focused stereotyped behaviour than repeated methamphetamine treatment. Stereotyped behaviour induced by methamphetamine plus scopolamine was reproduced by challenge injections of methamphetamine plus scopolamine, methamphetamine, and to a lesser extent by scopolamine challenges. The methamphetamine plus scopolamine-sensitized rats were conditioned to a low frequency tone (300 Hz, 100 dB) associated with the drug state. They exhibited a conditioned response to pairings of the tone (conditioned stimulus) and placebo injections. However, they did not respond to the tone alone or the placebo injections alone. The methamphetamine-sensitized rats failed to demonstrate any conditioning; only the repeated methamphetamine plus scopolamine treatment induced sensitization to the drug-associated tone. Pairings of exteroceptive conditioned stimulus-interoceptive unconditioned stimulus associations may provide an important source for conditioning to the tone associated with the drug state. We conclude that behavioural sensitization may operate via a reciprocal balance between the dopaminergic and cholinergic inhibitory systems, in favour of a dopaminergic dominance. Conditioning to the drug-associated tone may be mediated via a reciprocal balance between the two transmitter systems.     

MUC-1 mucin expression in invasive areas of intraductal papillary mucinous tumors of the pancreas

The expression of MUC-1 mucin (membrane-associated mucin) and MUG2 much (secretory mucin) were immunohistochemically examined in 46 invasive ductal carcinomas (IDC) and 16 intraductal papillary mucinous tumors (IPMT) of the pancreas. lntraductal papillary mucinous tumors usually reveal expansive growth. However, of the 16 IPMT examined in the present study, three showed an invasive growth pattern, which was similar to 'mucinous carcinoma', around the non-invasive growth areas. Of 46 IDC, MUCl much detected by monoclonal antibodies, DF3 and MY.1E12, was expressed in 44 cases (96%) and in 45 cases (98%), respectively, whereas MUC-2 mucin detected by polyclonal antibody, anti-MRP, was not expressed in any of the cases (0%). In contrast, in the non-invasive growth areas of the 16 IPMT, MUG1 much detected by DF3 and MY.1 E12 was expressed in four cases (25%) and in six cases (38%), respectively, whereas MUG2 mucin detected by anti-MRP was expressed in 13 cases (81%). The invasive growth areas of the three IPMT showed positive expression of MUG-1 mucins detected by DF3 and MY.1E12, although the non-invasive growth areas showed negative expression of MUG1 muclns, except for their focal positive expression in one of the three cases. These findings indicate that the invasive growth areas of IPMT acquire a characteristic of MUC-1 much expression that is usually seen In IDC.     

Real-time monitoring of HCV-RNA by single tube assay kit and potential importance for predicting virological sustained response in patients with chronic hepatitis C

In an attempt to predict virological sustained responders among patients with chronic hepatitis C after interferon therapy, HCV-RNA in serum was measured by a one tube RT-PCR assay kit using the RNA corresponding to 5 μL serum (standard assay) or 300 μL serum (enhanced-sensitivity assay). Dilution analysis revealed that sensitivity of the ‘enhanced-sensitivity assay’ increased by 10–100-fold when compared with a ‘standard assay'. Using these assays, prospective study of interferon therapy on 38 HCV-RNA seropositive cases with chronic hepatitis (total amount 702 MU; duration of treatment 5–6 months) was performed. At the end of treatment, six were still positive and 32 became negative by the ‘standard assay', whereas an additional eight cases became positive (total 14 cases positive; the remaining 24 cases negative) by the ‘enhanced-sensitivity assay'. Hepatitis C viral RNA state at the end of treatment remained the same 6 months later in 23 cases (61%) by a ‘standard assay’ and in 31 (82%) by the ‘enhanced-sensitivity assay'. Of importance was that all patients (14 cases) demonstrating HCV-RNA in serum at the end of therapy, even by the ‘enhanced-sensitivity assay', did not show the disappearance of HCV-RNA in serum despite the long follow up. From these results, in order to improve our treatment efficacy, we should try to modify our treatment protocol to the extent that at least HCV-RNA becomes undetectable. That can be only feasible during treatment by real-time monitoring of HCV-RNA.