Search

To elucidate the relationship between embryo grade and ART outcomes and determine how to decrease multiple pregnancy rates by assigning patients to single embryo transfer (SET) instead of dual embryo transfer (DET) according to embryo grade and/or availability.Retrospective medical record review.IVF fertility center.247 women undergoing day 5 DET after in vitro fertilization or intracytoplasmic sperm injection treatment.We retrospectively investigated embryo grade and outcomes on day 5 DET and calculated theoretical multiple pregnancy rates by assigning patients to SET instead of DET according to a combination of embryo grades and availability.Implantation, pregnancy, multiple pregnancy, expected pregnancy and expected multiple pregnancy rates.Embryo grade affects implantation rates so that embryo transfer of at least one embryo with gradeor = 3BB resulted in high multiple pregnancy rates as well as high pregnancy rates. By performing DET, the clinical pregnancy rate was 39.9% with a multiple pregnancy rate of 33.3%; however, had SET been performed with an embryo graded 3BB or better, theoretical calculated pregnancy rates would have dropped to 35.8% but with a multiple pregnancy rate of 7.2%.Our model showed that cases having at least one embryo with gradeor = 3BB might serve as suitable candidates for SET.

The role of resistin in human biology remains uncertain. We measured serum resistin levels in Japanese patients with (n=111) and without (n=98) type 2 diabetes mellitus and investigated the significance of this hormone in the pathophysiology of diabetes. The levels of serum adiponectin and leptin were also measured. Resistin levels were increased significantly in patients with type 2 diabetes compared with non-diabetic subjects (24.7+/-2.6 vs. 15.0+/-1.2 ng/ml, p=0.0013). However, there was no correlation in either patient group between serum resistin levels and markers of insulin resistance, obesity or hyperlipidaemia. These results were in direct contrast to the data of leptin or adiponectin, both of which were closely related to these clinical markers of diabetes. Multivariate regression analysis on the combined data of the two groups demonstrated that the presence of diabetes and HDL cholesterol levels were significant predictors of serum resistin levels (diabetes: beta=0.159, p=0.035; HDL: beta=-0.172, p=0.039). No correlation was observed between C-reactive protein and resistin adjusted for BMI. Taken together, these findings demonstrate that serum resistin levels are increased in patients with type 2 diabetes, but this increase is not linked to markers of insulin resistance or adiposity. Further studies are necessary to elucidate the significance of serum resistin concentration in human pathophysiology.

To evaluate the influence of aging on bronchial hyperreactivity (BHR) during the childhood period, age-related changes in bronchial reactivity to methacholine (BRm) in children from 2 to 13 yr of age were studied using the same method, employing a methacholine inhalation challenge with transcutaneous oxygen pressure (tcPO2) monitoring. Three hundred and thirty-nine asthmatic subjects (male:female = 200:139, aged [mean +/- SEM] 7.2 +/- 0.2 yr) and 107 age-matched controls (male:female = 55:52, aged 5.3 +/- 0.3 yr) were enrolled in this study. TcPO2 was measured by a tcPO2 monitor, and subsequent doses of methacholine were then doubled until a 10% decrease in tcPO2 from its baseline value was reached. The cumulative dose of methacholine at the inflection point of tcPO2 (Dmin-PO2) was considered to represent the BRm. For the sake of comparison, respiratory resistance (Rr) in subjects more than 6 yr old was measured by the oscillation technique during methacholine inhalation challenge, and the threshold point of Rr (Dmin-Rr) was also considered to represent the BRm. In the asthmatic children aged 2 to 7 yr, Dmin-PO2 decreased significantly from 12.2 +/- 2.1 to 3.1 +/- 0.8 units, but after age 8 yr the values gradually increased from 3.1 +/- 0.7 to 6.4 +/- 1.6 units. In children aged 6 to 13 yr, Dmin-Rr showed the same increase as Dmin-PO2, from 2.0 +/- 0.5 to 5.8 +/- 1.4 units. These age-related changes in BRm reflected in both Dmin-PO2 and Dmin-Rr were also seen in the age-matched controls.(ABSTRACT TRUNCATED AT 250 WORDS)

The extent to which respiratory muscles are exerted during partially supported ventilation is difficult to differentiate, because these muscles and the ventilator work simultaneously to produce ventilation. We have developed a new method for determining the pressure developed by the respiratory muscles in partially supported ventilation. In seven patients on pressure-support ventilation (PSV), pressure, flow, and lung volume change were measured at the airway opening. Various PSV levels (0–15 cmH2O) were applied to each patient in random order. By utilizing a model of respiratory mechanics, we calculated the pressure developed by the respiratory muscles and the inspiratory work performed by the muscles from the measured parameters by use of the resistance and elastance of the respiratory system obtained during controlled ventilation. Increasing PSV from 0 to 15 cmH2O modulated the resultant breathing pattern, i.e., increasing tidal volume and decreasing respiratory rate. The respiratory muscle pressure, although less negative, had a shape that corresponded to the shape of airway occlusion pressure at each PSV level, and both pressures decreased concomitantly with increasing PSV. The respiratory muscle work progressively decreased with increasing PSV. This analysis enabled clear and continuous quantifications of the respiratory muscle force generation and inspiratory work during partially supported ventilation.

We performed surgery for distal arch aneurysm in 22 patients; through a mediansternotomy in 11 patients (group M), and through a left thoracotomy approach in 11 patients (group L). If the distal position of an aneurysm was located at the pulmonary hilus without severe calcification of the ascending aorta, the mediansternotomy approach was chosen, whereas if we suspected that distal anastomosis would be difficult due to severe calcification of the descending aorta or aortic dissection type B, the left thoracotomy approach was selected. The mean cardiopulmonary bypass time (min.) was 229 +/- 56 in group M and 225 +/- 47 in group L, the mean circulation arrest time (min.) was 39 +/- 19 in group M and 31 +/- 5 in group L, and the mean lowest temperature (degree C) was 22 +/- 1 in group M and 20 +/- 2 in group L. No severe cerebral or cardiac complications developed in any patients. The hospital mortality was 9% (1/11) in group M and 9% (1/11) in group L. Following discharge, 8 (73%) patients from group M and 9 (92%) from group L resumed a normal lifestyle. There were no significant differences between the 2 groups and the operative results of both approaches proved satisfactory. Therefore our indications for each surgical approach to treat distal arch aneurysm seemed appropriate.

Multicystic dysplastic kidneys (MCDK) and obstructive renal dysplasia (ORD) are two different phenotypes of dysplasia commonly associated with urinary tract obstruction. However, the mechanisms whereby obstruction in the developing kidney leads to each dysplasia are unknown. In the present study, 16 fetal MCDKs and 3 fetal ORDs (18-35 weeks of gestation) were analyzed with light microscopy, point-counting morphometry, immunohistochemistry with a podocyte marker, and scanning electron microscopy. Additionally, reconstructions of dysplastic nephrons were done via serial section analysis. Early stages of MCDK and ORD similarly revealed numerous cyst formations, predominantly in the subcapsular nephrogenic zone. Occasionally, glomerular tuft remnants with mature podocyte phenotypes were observed in cysts, suggesting the acquisition of filtration. Three dimensionally, basic nephron structures were installed in the cystic nephrons, namely the macula densa with a primary loop structure. Cysts developed in the once-induced nephrons due to fluid retention in both MCDK and ORD. In utero urinary tract obstruction may cause urine retention in functioning nephrons and lead to glomerular cysts in the nephrogenic zone. These findings were common to MCDK and ORD, albeit at different sites of obstruction. Expansion of glomerular cysts with tubular dilatation (cysts) disturbs the subsequent nephrogenesis and may contribute to the misdevelopment of fetal kidneys.

Intracytoplasmic sperm injection (ICSI) has been validated as a useful treatment in severe male-factor patients who could not achieve fertilization and live births by conventional in vitro fertilization treatment. To examine the impact of male factors on ICSI outcome, clinical laboratory data were retrospectively analyzed.One hundred two cycles of ICSI treatment indicated by severe male-factor infertility were entered into this study. Sperm parameters including sperm motility, sperm concentration, and sperm motility index assessed by the Sperm Quality Analyzer were evaluated.Five hundred seventy-six metaphase II oocytes retrieved were manipulated. The normal fertilization (2 PN) rate per oocyte was 64.9 +/- 26.0% (mean +/- SD). Of the 99 transfers, 31 clinical pregnancies were obtained, yielding an average pregnancy rate of 31.3% per transfer. The mean sperm motility, sperm concentration, and sperm motility index were 20.3 +/- 16.1% (range, 0 to 50%), 18.2 +/- 25.1 x 10(6)/ml (range,1 to 150 x 10(6)/ml), and 31.2 +/- 45.0 (range, 0 to 220), respectively. Sperm concentration did not have a significant impact on fertilization rate by ICSI. In four cases, ICSI was performed using totally immotile sperm and the fertilization rate was 43.5%, which was significantly lower than that of some of the other sperm motility groups, and no pregnancy could be achieved. In 14 cases in which the sperm motility index assessed by the Sperm Quality Analyzer was 0, the fertilization rate (50.0%) was significantly lower than in most of the other sperm motility index groups.These findings suggest that in severe male-factor cases with totally immotile sperm or a sperm motility index of 0, the selection of good-quality sperm should be verified before injection.

To evaluate the reproducibility of the Ultrafast CT (UFCT) scoring system and assess its usefulness in monitoring clinical severity in infants with bronchopulmonary dysplasia (BPD).UFCT scoring was done in 22 infants (15 boys and 7 girls aged 1 to 37 months) with BPD. A total of 258 lung fields were evaluated for the presence of hyperaeration, linear opacities, triangular subpleural opacities, and bronchovascular bundle distortion or thickening, and UFCT scores were given. Intraobserver and interobserver agreement and reproducibility of UFCT scores were statistically analyzed. In 12 patients, UFCT scores were linearly correlated with clinical severity scores based on respiratory dysfunction and complexity of care."Hyperaeration," which was the most frequent (18 of 22, 81.8%) finding, showed high concordance (kappa = 0.73, p0.001, kappa = 0.59, p0.001), and its UFCT scores significantly correlated with intraobserver and interobserver analyses (r = 0.94, p0.001, r = 0.82, p0.001, respectively). UFCT scores for hyperaeration significantly correlated with clinical scores (r = 0.75, p0.01), whereas those for the others did not.UFCT is useful for assessing BPD. Hyperaeration was the most common and reproducible finding, and its extent significantly correlated with clinical severity.

The blocking effects of complement-dependent sperm immobilizing antibodies in the sera of infertile women and monoclonal antisperm antibodies against humans and mice on fertilization were investigated. The hemizona assay (HZA) and sperm penetration assay (SPA) were used to study the inhibitory effects of sera from 22 infertile patients positive for sperm immobilizing antibodies. Use of these tests allowed us to differentiate whether the antibody blocked sperm-zona pellucida tight binding and/or sperm penetration into the ooplasm. The zona pellucida penetration assay (ZPA) was also used to study the effects of four monoclonal antibodies (mAbs) on human sperm penetration into the zona pellucida. Seven mAbs against murine spermatozoa were tested for their inhibitory effects on in-vitro fertilization (IVF) and HZA in mice. Of 22 patient sera with sperm immobilizing antibodies, 21 (95.5%) inhibited HZA attachment and penetration, whereas this did not occur in any of 13 patient sera without these antibodies. However, 19 of 22 (86.4%) patient sera with sperm immobilizing antibodies and eight of 13 (61.5%) patient sera without these antibodies inhibited the SPA. Two (2C6, 1G12) of four mAbs against human spermatozoa showed strong inhibitory effects in all the assays (HZA, ZPA and SPA). One mAb (3B10) did not inhibit HZA but blocked ZPA and SPA. Another mAb (H6-3C4) seemed to have no inhibitory effects on fertilization. Two (Vx 5 and Vx 8) of seven mAbs against murine spermatozoa inhibited IVF in mice but did not block mouse HZA. These findings suggest that antisperm antibodies block fertilization at specific stages. Some of them may inhibit sperm capacitation and thus prevent all processes of fertilization that follow. Some other antibodies may not affect capacitation and sperm binding to zona pellucida but inhibit the acrosome reaction, followed by the blocking of sperm penetration through zona pellucida and ooplasm.

Between February 1988 and March 1993, 24 patients with locally advanced bladder cancer (stages T2-4N0-3M0) were included in this study. Combination chemotherapy consisting of methotrexate, vinblastine, epirubicin (doxorubicin) and cisplatin (M-VAC) was given to the patients in a neoadjuvant setting. The clinical stage was T2N0M0 in eight patients, T3aN0M0 in three, T3bN0M0 in seven, T4N0M0 in five and T4N3M0 in one. After chemotherapy, total cystectomy was performed in 20 patients and partial cystectomy 4. Of 24 patients, one (4%) showed a pathological complete response, and eight (33%) had a pathological partial response, for an overall response rate of 38% (95% confidence limits 20 to 57%). Nine patients who achieved a pathological response to chemotherapy had a significantly higher survival rate than the nonresponders (p0.01). In the relationship between the clinical stage and the response to chemotherapy, clinical T2 and T3a diseases were more likely to respond to chemotherapy than clinical T3b and T4 diseases, with a response rate of 64% and 15%, respectively. While a positive relationship between the pathological response and survival was observed, adequate follow-up is needed to assess the ability of neoadjuvant chemotherapy to improve the prognosis of patients with locally advanced bladder cancer.

From July 1993 through March 1994, 512 ESWL therapy was performed on upper urinary tract calculi in 277 renoureteral units using an EDAP LT-02 Lithotripter. Of these cases, 244 renoureteral units which were followed up for more than 3 months after ESWL were examined. These included 124 renal stones and 100 ureteral stones. Complete removal or sufficient stone disintegration was obtained in 66.7, 98.0 and 94.1% in the R1, R2 and R3 groups of renal stones, respectively, and in 92.4, 87.5 and 100% in the U1, U2 and U3 groups of ureteral stones, respectively. The over-all stone-free and success rates were 78.2 and 95.2%, respectively for renal stones and 93.0% and 94.0% respectively for ureteral stones. These results were better than those obtained by LT-01 in our experiences. The newly built-in coaxial X-ray system greatly contributed to these high success rates. As side effects, there were no serious complications. It is concluded that ESWL using an EDAP LT-02 is safer and more effective for the treatment of upper urinary tract calculi.

The diuretic furosemide has been reported to have a protective effect on allergic asthmatic reactions. This study was performed to investigate the effect of aerosolized furosemide on the lung clearance of 99mTc-diethylene triamine pentaacetic acid (99mTc-DTPA).Pulmonary clearance rates of 99mTc-DTPA were measured by a computerized gamma camera with and without the inhalation of aerosol furosemide in 6 nonsmoking normal volunteers (Group 1), 7 smokers without pulmonary disease (Group 2) and 11 patients with asthma (Group 3).None of the six normal volunteers showed significant effects of inhaled furosemide on the 99mTc-DTPA clearance rates. Three of seven smokers presented an accelerated 99mTc-DTPA clearance by inhaled furosemide and the other four showed no significant change of 99mTc-DTPA clearance by furosemide inhalation. However, in 10 of 11 patients with asthma, there was significant suppression of 99mTc-DTPA clearance by furosemide inhalation.Asthmatics possess a furosemide-sensitive mechanism. Pulmonary aerosol scintigraphy with 99mTc-DTPA will be useful in predicting the effect of inhaled furosemide therapy.

To evaluate alveolar epithelial damage in PSS, we studied pulmonary epithelial permeability by measuring the clearance of inhaled 99mTc-DTPA aerosol and performing thin slice CT scan, pulmonary function tests and right heart catheterization in 28 patients with PSS. The 99mTc-DTPA clearance rate (kep value) in PSS was greater than in 11 non-smoking normal subjects (18.2 +/- 7.63 x 10(-3)/min vs. 9.12 +/- 0.77 x 10(-3)/min, p less than 0.01). In PSS, the kep value did not correlate with age, sex, duration of illness, dermal lesions, % vital capacity, or PaO2. In contrast, the kep value showed significant correlations with %DLco (diffusing capacity for carbon monoxide), extent of interstitial lesions evaluated by CT scan (CT score), and mean pulmonary artery pressure. On the other hand, the kep value was high in some patients with normal CT scan and normal %DLco. These findings indicate that pulmonary interstitial lesions in PSS are accompanied by alveolar epithelial damage, and that the clearance of 99mTc-DTPA may be an early predictor of interstitial change.

We reported 11 cases of patients who developed moderate to severe anaphylactic reactions induced by the ingestion of grand keyhole limpet (GKL) and abalone. Specific IgE-mediated hypersensitivity to these shellfish was demonstrated by history, skin prick test, RAST and immunoblotting. The RAST inhibition technique revealed cross antigenicity between GKL, abalone and keyhole limpet hemocyanin. By immunoblotting analysis, the major antigens of GKL are shown in the MW range of about 38 Kd and 80 Kd.

We studied the immunoglobulin levels and the number of eosinophils in the peripheral blood during the course of Mycoplasma pneumoniae pneumonia in children. Nineteen children with Mycoplasma pneumoniae pneumonia, including eight asthmatic children, were the study subjects. In eight of them, bronchial hypersensitivity was also measured by the Astograph method in both the convalescent phase and the healing (non-symptomatic) phase. The results were as follows. 1) Total serum IgA and IgM increased in the convalescent phase. 2) The levels of total serum IgE were higher in the acute phase and then decreased gradually. 3) The number of eosinophils in the peripheral blood increased over the acute-to-convalescent phase. 4) Bronchial hypersensitivity increased after Mycoplasma pneumoniae pneumonia, and two asthmatic children who exhibited bronchial hypersensitivity had asthmatic attacks. 5) Bronchial hypersensitivity persisted for over one month in two non-asthmatic children.

Airway inflammation with polymorphonuclear leukocytes (PMN) may play an important role in bronchial hyperresponsiveness (BHR). PMN generate superoxide anion (O2-) and other oxygen radicals that can damage lung tissue. We investigated the ability of peripheral PMN of children with bronchial asthma and control subjects to generate O2- and other active oxygen species using a 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-a]pyrazin++ +-3-one, a highly sensitive and specific chemiluminescence (CL) probe for O2-, and luminol-dependent CL. The ability of PMN of subjects with asthma to generate O2- and other active oxygen species was significantly greater than that of PMN of control subjects when stimulated with opsonized zymosan (OZ), phorbol myristate acetate or N-formyl-methionyl-leucyl-phenylalanine. Furthermore, in the same asthmatic children, the generation of O2- and other active oxygen species was significantly higher with attacks than without attacks when PMN were stimulated with OZ. We also demonstrated that O2- generation correlated with the degree of BHR to inhaled histamine. These results suggest that PMN of asthmatic children, especially those with attacks, generate more active oxygen species than that of control subjects and that airway inflammation caused by O2- may be closely related to BHR in subjects with bronchial asthma.

The effects of pH and osmolarity on cough receptors were evaluated by inhalation of aqueous aerosol in 37 children with bronchial asthma. Each of 3 different pH solutions (low 1.85-3.25, neutral 6.9-7.1, high 8.0-8.4) was combined with 3 different osmolarities (hypo 0-31, iso 254-292, hyper 830-1117 mOsm/kg), and 9 solutions were prepared. To evaluate the effect of low chloride ion concentration, neutral pH and iso-osmolar solution including low chloride ion was also prepared. These aqueous aerosols were administered from a Devilbiss 646 nebulizer for 10 seconds. Coughing was induced in 28 subjects (76%) by inhalation of low pH and hyperosmolar solution, in one subject (3%) by low pH and iso-osmolar, in one (3%) by low pH and hypo-osmolar, in three (8%) by neutral pH and hypo-osmolar, and in none by any of the other solutions. It was suggested that the alteration of pH or osmolarity alone could not induce coughing effectively, and that the combination of pH and osmolarity was the important factor for the induction of coughing.

We report lysinuric protein intolerance (LPI) and systemic lupus erythematosus (SLE) in a 14-year-old girl. The co-existence of LPI and SLE in the same patient has been reported before and suggests that SLE is directly connected with LPI. Conclusion Possibly in LPI the metabolic derangement can cause immunological abnormalities. Therefore, when multisystem involvement is observed in LPI patients, SLE should be considered as a rare complication.

Alloreactive and soluble antigen-reactive, I-A-restricted T cell clones were examined for their ability to recognize hybrid I-A antigens. Several clones that recognized hybrid I-A(b)/I-A(k) molecules on (C57BL/6 x A/J)F(1) [(B6A)F(1)] spleen cells were studied. We were able to distinguish clones that recognized hybrid I-A molecules of the A(b)(a)A(k)(beta) type from those that recognized A(k)(a)A(b)(beta) molecules. We reached this conclusion by considering data from three independent types of experiments. (a) Monoclonal antibodies were used to inhibit T cell stimulation. Antibodies 10.2.16 and H116.32 distinguished two mutually exclusive "families" of T cell clones. One group of clones was inhibited by 10-2.16 and not H116.32, the other group exhibited reciprocal inhibition. (b) T cell proliferation was assayed using antigen-presenting cells from B6.C-H-2(bml2) (bml2) and [bml2 x B10.A(4R)]F(1) mice. Because the bml2 strain has a mutation that results in an altered A(b)(beta) polypeptide chain (A(bm12)(beta)), we reasoned that clones that could recognize the [bm12 x B 10.A(4R)]F(1) cells were recognizing A(b)(a)A(k)(beta) molecules. Alternatively, clones not recognizing [bml2 x B10.A(4R)]F(1) cells had specificity for A(k)(a)A(b)(beta) molecules. (c) I-A molecules immunoprecipitated from radiolabeled (B6A)F(1) splenocyte extracts were analyzed by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These experiments confirmed an earlier report that antibody 10.2.16 recognized determinants on the A(k)(beta) chain (12). Antibody H116.32 immunoprecipitated products consistent with recognition of A(k)(a) determinants. Taken together, these three types of results offer conclusive evidence that T cell clones recognizing "hybrid" I-A molecules use either A(b(k)A(k)(beta) or A(k)(a)A(b)(beta) molecules as recognition or restriction sites. Clones whose proliferation was supported by [bm 12 x B10.A(4R)]F(1) cells and blocked by anti-I-A(k) antibody 10-2.16 recognized A(b)(a)A(k)(beta) B molecules. Clones that were blocked by antibody H116.32 and did not recognize [bml2 X B10.A(4R)]F(1) cells use a recognition site(s) on A(b)(a)A(k)(beta) molecules. Thus, we can demonstrate both functionally and biochemically that hybrid F(1) I-A molecules of the structure A(k)(a)A(b)(beta) and A(b)(a)A(k)(beta) both exist on (B6A)F(1) splenocytes and that both configurations are used in immune recognition phenomena.

This is a case report of a successful case of surgical treatment of a lung abscess formed from pulmonary aspergilloma in a patient with a recurrence of acute promyelocytic leukemia during remission induction therapy. A 39 year-old woman was admitted to our hospital for remission induction therapy of a recurrence of acute promyelocytic leukemia. We were successful in bringing her to a state of complete remission by a therapy of a intermediate-dose of Ara-C. Pulmonary aspergilloma was formed in the right lung during therapy. We performed transbronchial infusions to Amphotericin B which dissipated the fungus ball. However, a lung abscess was formed. We needed radical therapy, and there were no complications in the following surgical treatment. Recently, along with the tendency for the use of stronger chemotherapy treatment regimes for acute leukemia, there has been an increase of the number of mycosis cases such as the pulmonary aspergilloma case reported here. Accordingly, surgical treatment is becoming more and more a matter of necessity in the clinical course of leukemia patients.

For quantitative estimation of sperm immobilizing antibody, a new assay method was developed and applied to a follow-up study of antibody titers in the sera of sterile women. The 50% sperm immobilization units (SI50) estimated by the quantitative method ranged from 1.2 to 97.3 in the sera from 16 sterile women who showed relatively high antibody activities in the previous semiquantitative sperm immobilization test. When the quantitative antibody titers, SI50, were followed over 3 years in sterile women with the sperm immobilizing antibody, the antibody titers were found to be unstable, and undulated over a period of several months. Therapy consisting of use of a condom seemed to be ineffective in decreasing the antibody titers.

A case of mixed infection of Salmonella (type C2) and Parasarcophaga crassipalpis (an intestinal myiasis) in a two-year-old girl is reported. A girl after dining in a restaurant developed typical Salmonella food-poisoning symptoms. Stool culture showed both Salmonella and fly larvae. This paper is to report a first case of this fly larvae infection and to review the recent trends of food-poisoning in Japan.

Monoclonal antibodies (mAb) were used to inhibit the proliferation of antigen-reactive (C57BL6/J X A/J)F1 restricted T cell clones. We have been able to subdivide these F1 restricted T cell clones into two groups: one of which recognizes the A alpha k A beta b molecule and the other group which recognizes the A alpha b A beta k molecule. Using clones with defined reactivities, we could assign the reactivities of monoclonals to the A alpha or A beta chains. By immunoprecipitation and two-dimensional analysis of Ia molecules from F1 spleen cells, we could independently map the reactivities of the mAb as being determined by the A alpha or A beta chain. To date, these two methods of chain localization of the antibody reactivity have agreed. Further, the differential blocking of the A alpha k A beta b restricted T cell clones suggests that there exists more than one restriction site per Ia molecule. Increasing the number of possible functional Ia restriction sites, either through combinatorial association of alpha and beta chains or by using more than one site per molecule, should increase the number of ways Ia molecules can function in antigen presentation.

Thirteen infertile patients (18 cycles) with sperm immobilizing antibodies were subjected to in vitro fertilization and embryo replacement (IVF-ER) therapy during 12 months from January to December 1985. Four patients became pregnant, two of them delivered healthy babies at term and 2 had abortions at 5 weeks and 12 weeks of gestation, respectively. In the same duration, 24 patients (28 cycles) with tubal factor and 4 patients (5 cycles) with male factor for infertility were subjected to IVF-ER, and two patients with tubal factor became pregnant and delivered healthy babies at term. In the patients with immunological factor, fertilization and cleavage rates per mature oocyte were 85.9% (55 fertilized/64 oocytes) and 81.3% (52 cleaved/64 oocytes) respectively, while the fertilization rates in patients with tubal factor and male factor were 70.5% (62 fertilized/88 oocytes) and 50.0% (6 fertilized/12 oocytes) respectively and all fertilized eggs in these patients developed to the cleavage stage. Thus fertilization and cleavage rates for mature oocytes from the patients with the immunological factor were slightly better than those with the tubal factor and much better than those with the male factor. The antisperm antibody titers (SI50) in sera determined by the quantitative sperm immobilization test ranged from 20 to 243 units while those in follicular fluids ranged from 21 to 160 units in the patients with the immunological factor. The follicular sperm immobilizing antibodies could be detected in any patient who had the antibodies in the serum. Immunoglobulin (IgG, IgA, IgM) concentrations in follicular fluids were not significantly different from each other in the patients with immunological, tubal and male factor for infertility.

Since anti-sperm antibodies were first discovered in the sera of women, the relationship of these antibodies to sterility has been studied by many investigators. In order to determine the antigens of spermatozoa responsible for raising antibodies to spermatozoa in humans, many studies have been carried out by purifying human spermatozoa cell membrane and seminal plasma components. Since it was found that the purification was difficult by physiochemical procedures, the immunoaffinity chromatography bound monoclonal antibody (Mab) to spermatozoa antigens was attempted for this purpose. The establishment of hybridomas producing Mabs to human seminal plasma and human spermatozoa was reported by Shigeta et al. (1980), Isojima, Koyoma & Fujiwara (1982), Lee et al. (1982) and Isahakia & Alexander (1984). The ordinary approaches to obtain the Mabs consisted of xenogenic immunization with human semen and cell fusion of immunized spleen cells with mouse myeloma cells. However, the antigenic epitopes of human spermatozoa, which induced antibody production, are xenogenic for the mouse, and therefore there is a possibility that there is a difference in recognized antigenic epitopes in humans as isotypic and in mice as xenogenic. In order to study these antigenic epitopes, which correspond to antibodies against spermatozoa in women, the establishment of human-mouse hybridomas, which produced anti-semen antibodies as produced in sterile women, became essential. In these studies, we used recently developed cell fusion techniques to fuse immunized human peripheral lymphocytes with mouse myeloma cells.

There are several ways to make immunological contraceptive vaccines. We are trying to purify gamete antigens from human seminal plasma and porcine zona pellucida (ZP) by using several monoclonal antibodies (Mabs) for this purpose. A human seminal antigen relevant to sperm immobilization with corresponding antibody was purified by immunoaffinity chromatography-bound Mab 1C4. This antigen was common to human milk protein but is different from lactoferrin. It was termed "ferrisplan" and was secreted mainly from the epithelial cells of seminal vesicle and adhered to the midpiece and postnuclear cap region of spermatozoa. Another Mab 2C6 reacted only to the SP component but not to other organ extract including human milk protein. The corresponding antigen to Mab 2C6 is aimed to purify by using high performance liquid chromatography and immunoaffinity chromatography-bound Mab 2C6. Several Mabs to porcine ZP antigens common to human ZP did not block human in vitro fertilization unless the second antibody (anti-mouse gamma-globulin) was used. It is speculated that porcine ZP will not contain the antigenic component that relates to sperm receptor of human ZP. Therefore, an antigenic molecule located close to sperm receptor of human ZP must be purified from the porcine ZP by using immunoaffinity chromatography-bound Mab and other biochemical methods, expecting production of a corresponding antibody that blocks the fertilization in humans.

Advanced gastric cancer cases (42 cases) undergone gastrectomy were studied on the correlation between tissue uptakes and prognosis. The patients were administrated Tegafur preoperatively and tissue samples were obtained intraoperatively, 5-FU levels in tumor and lymph node were measured by GCMF. The patients, being measured 5-FU uptake by tissue and received Tegafur over 60 g as postoperative adjuvant chemotherapy, were divided into two groups; namely, one with 5-FU up take by tumor tissue and lymph node was higher than 0.05 mcg/g and the other with lower than 0.05 mcg/g. Both groups showed no significant difference in background factors. Each survival rate was calculated by Kaplan-Meier method and examined by generalized Wilcoxon method. As the result, there was no significant correlation in the 5-FU uptake by tumor and prognosis, but prolongation of survival rate in the group of 5-FU uptake by lymph node over 0.05 mcg/g was suggested.

Data presented in this paper demonstrate the existence of two separate pathways by which a single T cell clone can induce B cell differentiation. With the use of high doses of antigen, a T cell clone can induce a primary antibody response in unprimed B cells. With the use of low doses of antigen, the same T cell clone can induce an immunoglobulin (Ig)G response in primed B cells. The primary response is accompanied by T cell proliferation and lymphokine production (interleukin 2, B cell growth factor, B cell differentiation factor for immunoglobulin M, and B cell differentiation factor for immunoglobulin G). The secondary response does not require proliferation and occurs independently of detectable lymphokine production. Variants of the wild type T cell helper clone have been isolated. One variant can provide help to unprimed B cells when high doses of antigen are used. This variant cannot provide help to primed B cells when low doses of antigen are used, nor can it provide help to CBA/N "xid" B cells at any antigen concentration tested. Additional variants have been isolated that proliferate on antigen-pulsed-presenting cells, but fail to secrete detectable lymphokines and do not induce B cell differentiation. These results suggest that a single T cell helper clone has multiple functional activities that can be independently expressed.

In order to apply the early pregnancy factor (EPF) to early diagnosis of fertilization, the establishment of optimal conditions for assay of EPF was attempted, and then EPF in the sera of contracepted and conceived women 4 to 6 days after ovulation were measured. For assay of EPF, 0.25 ml of 1:2 step diluted anti-human lymphocyte serum (ALS), 0.05ml of guinea pig serum as complement and 0.1ml of lymphocytes suspension (1 X 10(7)/ml) pretreated with test serum were mixed and then incubated at 37 degrees C for 90 min. To this mixture 0.1ml of sheep red blood cell suspension (2 X 10(9)/ml) was added and the rosette formation was counted after centrifugation. The rosette inhibition titer (RIT) was expressed as reciprocal of ALS dilutions which resulted in less than 75% of rosette formation as compared with the control. RITs of the conceived women who were assayed on the 5 th day after ovulation were in the range from 16 to 32 X 10(3), while that of the control contraceptive women who were assayed on the same day was in the range from 2 to 4 X 10(3). The sterile women who received AIH but failed to conceive all showed less than 4 X 10(3) as RIT. These results suggest that the assay of EPF is valuable in detecting the early stage of fertilization and possibly may help to differentiate the impairment of embryo implantation from non-fertilization of the ovum as a cause of sterility.

Forty-one patients with advanced gastric cancer underwent gastrectomy, and the correlation between tissue uptake of the adjuvant drug and the prognosis were studied. The patients were preoperatively administered Tegafur and samples of tissue were obtained intraoperatively. 5-FU levels in the tumor and lymphnodes were measured by gas chromato-mass fragmentography (GCMF). The patients measured for 5-FU tissue uptake were given more than 60 g of tegafur as postoperative adjuvant chemotherapy, and divided into two groups; one in which the 5-FU uptake by tumor tissue and lymphnode was over 0.05 microgram/g and the other lower than 0.05 microgram/g. In both groups there were no significant differences in background factors. Each survival rate was calculated by the Kaplan-Meier method, and the generalized Wilcoxon method was used for statistical analysis. There was no statistically significant correlation between 5-FU uptake by the tumor and the prognosis, but the 5-year survival rate in the group with over 0.05 microgram/g uptake by lymphnodes was statistically significant (p = 0.018).