Immunogenicity and protective efficacy of orally administered recombinant Lactococcus lactis expressing surface-bound HIV Env

This study investigates whether genetically modified orally administered Lactococcus lactis (L lactis) could be used as an HIV vaccine. L lactis is immunogenic and extremely safe when delivered orally. We created a recombinant L lactis vector expressing the envelope protein of HIV on its cell surface. Oral immunization with this vector induced high levels of HIV-specific serum IgG and fecal IgA antibodies. Cell-mediated immune responses also were generated in both the regional lymph nodes and the spleen. Dendritic cells are readily infected by L lactis and appear to play a potential role in mediating the development of these immune responses. The protective efficacy of this vaccine strategy was demonstrated by challenging mice intraperitoneally with an HIV Env-expressing vaccinia virus. Their viral loads were 350-fold lower than those of control mice. These findings support the further development of L lactis-based HIV vaccines.     

The evaluation of ultrasonic oocyte collection techniques for IVF-ET--comparing the transabdominal and transvaginal methods

In order to evaluate the effectiveness and safety of ultrasonic oocyte collection for IVF-ET, a comparative study was performed on the results of IVF-ET by the transabdominal and transvaginal methods in 145 cycles of 51 patients. The oocyte collection rate for the transvaginal method was significantly higher than that for the transabdominal method (77.1% vs. 68.7% per follicle, p less than 0.01). The majority of the oocytes were collected in three follicular flushings. The fertilization rate for oocytes collected by the transabdominal method was significantly higher than that for those collected by the transvaginal method in cases of tubal damage and endometriosis (81.1% vs. 65.6% per oocyte, p less than 0.01). There was no significant difference between the signs of complications seen with the two methods. Six successful pregnancies were obtained by IVF-ET in our hospital (4 cases by the transabdominal method, and 2 cases by the transvaginal method). Two of these cases have already delivered full term healthy male infants. In conclusion, these results indicate that ultrasonic oocyte collection has been established as the most effective method for IVF-ET by means of either the transabdominal or the transvaginal methods.     

A unique case of subepidermal bullous disease

A 72-year-old Japanese woman developed small, asymptomatic, subepidermal vesicles on the limbs, back, chest and abdomen. Immunoperoxidase staining of the lesional skin showed linear deposition of IgG, IgA and C₃ along the basement membrane zone (BMZ), and indirect immunofluorescence studies revealed IgG and IgA class circulating anti-BMZ auto-antibodies in the patient's serum. Ultrastructurally, the vesicles were caused by dermo-epidermal separation at the lamina lucida, and the immune deposits were located just beneath the basal lamina in a band-like pattern. Immunoelectron microscopic observation of normal human skin incubated with the patient's serum using an organ culture system revealed that the anti-BMZ antibodies reacted with the anchoring fibrils. Administration of dapson was not effective, but betamethasone was. This case of subepidermal bullous disease is unique and cannot be classified into any existing category of bullous dermatoses.     

Prognostic significance of the bromodeoxyuridine labeling index in primary colorectal carcinoma

The bromodeoxyuridine (BrdU) labeling index was determined in 40 primary colorectal carcinomas by DNA flow cytometry using a BrdU-specific monoclonal antibody. The labeling index, or the fraction of cells in the S-phase of the cell cycle, ranged from 12% to 52%, with a mean of 28% (SEM, 2%). The labeling index in 19 patients was over 30%, which was termed a higher labeling index. There was no significant difference in the labeling index based on the clinical stage of the disease. During the 5-year follow-up after the apparently curative resection, 14 patients died of the disease, 1 died of an unrelated cause, 1 is alive with a recurrence of the disease, and 24 are alive without the disease. The higher labeling index was thus associated with a significantly poorer patient survival (P=0.03 based on the generalized Wilcoxon test). The present study therefore disclosed that the S-phase fraction of tumor cells thus determined had prognostic significance in primary colorectal carcinoma.     

Molecular cloning and expression of a dextranase gene from Arthrobacter in Streptococcus sanguis

The gene coding for a dextranase activity of Arthrobacter CB-8, named dex gene, was isolated and cloned into Escherichia coli and into Streptococcus sanguis. The gene library was screened by transparent halo formation around the colonies grown on agar medium containing blue dextran. DNA fragment consisting of about 3,200 base pairs was prepared for further cloning procedures. Dextranase activity was detected in the periplasmic space of E. coli clones, using pUC19, pVA 838 and their derivatives. Dex gene was also introduced into S. sanguis Challis using pVA 838, a plasmid that is able to replicate in both E. coli and S. sanguis. But the clones did not express the dex gene. For the expression of dex gene in S. sanguis, a new shuttle vector was constructed, which contained the promoter region of a glucosyltransferase gene from S. mutans as well as the terminator region of ribosomal RNA from E. coli. The plasmid was designated pMNK. Using pMNK as vector, dex gene was expressed in S. sanguis. Dextranase activity was detected in the cellular fraction of the clones.     

Venous return disturbances in chronic pulmonary disease: a study using pulsed Doppler echocardiography]

Pulsed Doppler echocardiography was used for investigation of respiratory changes of superior vena caval flow in chronic pulmonary disease to analyze the mode of venous return and its regulating factors. The subjects consisted of 55 patients with various pulmonary diseases and 44 healthy controls. 1. In the controls, the velocities of the S and D waves increased during the inspiratory phase. 2. Patients with pulmonary diseases were categorized into 2 groups. One with the mode of the healthy subjects (normal pattern), and the other with disappearance of the D wave or both the S and D waves (abnormal pattern) in the expiratory phase. All patients with restricted ventilation exhibited the normal patterns, while the abnormal patterns were observed in some of the patients with obstructive ventilation, and most of the patients with combined ventilation. 3. The respiratory patterns of the pressure gradients between the right atrium and subclavian vein reflected patterns of the superior vena caval flow well. In cases with abnormal patterns, the right atrial pressure exceeded the subclavian vein pressure in the expiratory phase. 4. The velocity of the A wave at the expiratory phase correlated significantly with pulmonary vascular resistance and with the mean pulmonary artery pressure. In conclusion, the mode of venous return in patients with pulmonary disease varies depending on the mode of the ventilation disturbance and the presence of right ventricular pressure overload, which is most likely caused by the intrathoracic pressure and by right ventricular filling abnormalities.