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Although apoptosis is believed to play an important role in the ontogenetic development of animals, the molecular mechanism that triggers the regression of liver hemopoiesis during the perinatal period is not known. Apoptosis is induced by many factors such as a decrease in growth factors and increased oxygen stress. Since hepatic gamma-glutamyl transferase (GT) levels change markedly during the perinatal period in rodents, the metabolism of glutathione (GSH), a naturally occurring major antioxidant, might change significantly in and around liver cells. Hemopoietic cells but not hepatocytes exhibit significant apoptosis in thiol-free medium and the hemopoietic apoptosis can be inhibited by various thiols, such as L-cysteine, N-acetyl-L-cysteine, and GSH. The contribution of GSH levels in and around fetal liver cells in the triggering of apoptosis in hemopoietic cells is discussed.

Aspect ratios increase as metal device design rules decrease and, in the current metal etching process, this higher aspect ratio has lead to increased electron shading (Hashimoto, 1993) which has been shown to cause electrical and physical damage (Tabara, 1996) to the device. In case of Al etching, it is very difficult to satisfy the device's requirements due to its own characterization. In poly etching, the electron shading effect can be reduced by replacing the photoresist mask with a SiO/sub 2/ hard mask (Lee et al., 1998). This same method has been applied to Al etching and has demonstrated similar results. In this paper, we show that using an SiO/sub 2/ hard mask instead of photoresist is effective in preventing electrical damage in Al etching.

Microscratches and particles are becoming a serious problem associated with the introduction of CMP to IC-processing. This paper describes a new surface inspection system for a blank wafer with emphasis on the discrimination technique between microscratches and particles. The system first detects both defects by means of the laser scattering technique as is the case of a conventional particle inspection tool, followed by a newly developed discrimination procedure based on the difference of angular distribution of the scattered light. The discrimination rate which is defined as the ratio of the correctly classified defects to the total defects detected was evaluated to be 85% or more on actual CMP-processed wafers.

A novel architecture for an SRAM-based user-reprogrammable gate array which consists of programmable logic elements (PLEs), switching stations (SSs), wirings, and input/output blocks (IOBs) is discussed. The SS is designed to connect its neighboring PLEs and/or IOBs through only one NMOS pass transistor to maintain a sufficient signal level and reduce the delay in signal propagation. Hidden interconnection networks which directly merge neighboring PLEs were adopted to expand the number of inputs and product terms of combinational logic, and allow the construction of a parallel-to-serial/serial-to-parallel converter without using SSs. A fabricated CMOS prototype chip includes an 8 Kbit 50 ns temporary storage memory which is programmable to the random access mode or the FIFO mode with a word length of 4 bit or 8 bit. >

A magnetic levitation transport (MLT) system in vacuum has been developed. The system can carry silicon wafers into a number of process chambers for LSI fabrication under particle-free conditions. Since there are no sliding mechanical parts in this system, it does not need lubricants. Therefore, the transport system is free from oil and has a long lifetime. We measured the contamination of dust particles on silicon wafers when the wafers were transferred in vacuum with the MLT system. In this paper, we describe the MLT system and present the results of dust particle measurements.

A 41-year-old Japanese woman complained of a gradually enlarging swelling of her left cheek for seven months. She was diagnosed with osteopetrosis by standard skeletal radiographs, and her cheek swelling was diagnosed as maxillary osteomyelitis secondary to osteopetrosis. She underwent a left partial maxillectomy, and her post-operative course was stable with no complications. A literature review is also presented.

We investigated the platelet aggregation inhibiting activity of human placental brush border membrane vesicles (BBMV) and obtained the following results. A strong platelet aggregation inhibiting activity existed in placental BBMV. The BBMV inhibited the platelet aggregation induced by ADP, arachidonic acid, collagen and ristocetin in a dose-dependent manner. The protein concentration of BBMV giving 50 per cent inhibition was 52 +/- 6 micrograms/ml for ADP-induced platelet aggregation, 21 +/- 2 micrograms/ml for arachidonic acid-induced platelet aggregation, 19 +/- 2 micrograms/ml for collagen-induced platelet aggregation and 107 +/- 9 micrograms/ml for ristocetin-induced platelet aggregation. There was a high level of ADP degrading activity (ADPase activity) in the placental BBMV. ADP degrading activity of the BBMV: 10.5 +/- 0.5 mumol/mg protein/min was 21 times greater than that of homogenate of the placental villi. The placental BBMV inhibited platelet TXA2 production. In the 40 micrograms/ml protein concentration of placental BBMV, platelet TXA2 production was almost completely inhibited.

We studied the platelet aggregation inhibiting activity and ADP degrading activity of human placental villi (tissue culture supernatant) and brush border membrane vesicles (BBMV) and obtained the following results. 1. There existed a platelet aggregation inhibiting activity in tissue culture supernatant of villi (S-villi) but not in that of decidua or amnion. The S-villi inhibited the platelet aggregation induced by ADP, but not that induced by collagen, arachidonic acid or ristocetin. And, there was also ADP degrading activity (ADPase activity) in the S-villi. ADP was quickly degraded by S-villi. When ADP was preincubated with S-villi, the platelet aggregation induced by ADP was completely lost. 2. There was very strong platelet aggregation inhibiting activity in placental BBMV. The BBMV almost completely inhibited the platelet aggregation induced by ADP, collagen, arachidonic acid and ristocetin. And there was very strong ADP degrading activity in the placental BBMV. ADP was quickly degraded by BBMV. When ADP was preincubated with BBMV, the platelet aggregation induced by ADP was completely lost. 3. The enzymatic character (heat stability, enzymatic kinetics, Ca++ dependency and pH dependency) of ADP degrading activity in BBMV was very similar to that in S-villi. 4. The ADP degrading activity of both S-villi and solubilized BBMV were fractionated by anion exchange column chromatography and gel filtration column chromatography in similar patterns, and it was shown that ADP degrading substance of both S-villi and solubilized BBMV had a molecular weight of about 60K.

To clarify the role of placental villi in maintaining microcirculation in the placenta, we investigated the action to protect the activity of prostacyclin in the tissue culture supernatant of human placental villi. 1. The platelet aggregation inhibiting activity of prostacyclin was decreased very rapidly in Ringer buffer. 2. The tissue culture supernatant of human placental villi has an action which protects the activity of prostacyclin. On the other hand, in the tissue culture supernatant of human placental decidua no such action is prominent. 3. Human chorionic gonadotropin (hCG) also has an action which protects the activity of prostacyclin. These results indicated that human placental villi play an important role in maintaining placental microcirculation by producing substances which have a protective on the activity of prostacyclin.

During the prenatal period the ability of the fetus mononuclear cells (MNC) to produce antibodies is impaired. 1. In term infants, IgG production with only Staphylococcus aureus Cowan I strain (SAC) stimulation was significantly decreased (p less than 0.00005) as compared with adults. However, when adult MNC supernatant stimulated with Pokeweed mitogen (PWM-sup) was given to full term infants, antibody production significantly increased. 2. The activity inducing antibody production in the full term infant MNC is present primarily in the adult T cell culture supernatant. 3. B cell differentiation factor gamma (BCDF gamma) and BCDF mu, which induce IgG and IgM production, respectively, were significantly reduced in premature and full term infants (p less than 0.00005) as compared with adult. 4. By giving adult PWM-sup to the under 33-week-old infants MNC with SAC stimulation, the induction of both IgG and IgM was reduced, and IgG production was especially inadequate. From the above, it is thought that in full term infants, the production of BCDF gamma and BCDF mu which are produced by T cells is reduced compared to that of adults, and that this is mainly responsible for the reduced antibody production. In addition, in the under 33-week-old infants, the immaturity of B cells and inadequate BCDF production induce underlying poor or inadequate immunoglobulin production.

To clarify the role of vitamin E (alpha-tocopherol) during pregnancy, we mainly investigated the relation between platelet function and vitamin E and obtained the following results. 1. The concentration of vitamin E (alpha-tocopherol) in plasma and platelet increased gradually during pregnancy. The concentration of vitamin E in plasma was 6.65 micrograms/ml in non pregnant women and 15.5 micrograms/ml in full term pregnant women. And the concentration of vitamin E in platelet was 99 micrograms/g protein in non pregnant women and 244 micrograms/g protein in full term pregnant women. 2. Vitamin E (alpha-tocopherol) inhibited platelet aggregation induced by ADP (adenosine diphosphate) and PAF(platelet activating factor). At a concentration of 500 micrograms/ml vitamin E almost completely inhibited platelet aggregation induced by ADP and PAF. 3. There exists a strong platelet aggregation inhibiting activity in human placental brush border membrane vesicles. At a protein concentration of 20 micrograms/ml the brush border membrane vesicles almost completely inhibited platelet aggregation induced by ADP and PAF. 4. With the co-existence of the brush border membrane vesicles, the platelet aggregation inhibiting activity of vitamin E (alpha-tocopherol) increased prominently. These results indicate that vitamin E plays an important role in the regulation of platelet function and might contribute to the maintenance of placental microcirculation by inhibiting platelet aggregation.

We studied the platelet aggregation inhibiting activity of human placental syncytiotrophoblast brush border membrane vesicles (BBMV) and obtained the following results. 1. There existed a very strong platelet aggregation inhibiting activity in placental BBMV. The BBMV almost completely inhibited the platelet aggregation induced by ADP, arachidonic acid and ristocetin. 2. There existed a very strong ADP degrading activity (ADPase activity) in the placental BBMV. ADP was quickly decomposed to AMP by BBMV. 3. Placental BBMV inhibited platelet TXA2 (TXB2) production. In the 0.04 mg/ml protein concentration of BBMV, platelet TXA2 production was almost completely inhibited. 4. There existed a very strong platelet aggregation inhibiting activity in solved BBMV. Soluble BBMV almost completely inhibited the platelet aggregation induced by ADP and arachidonic acid but did not inhibit the platelet aggregation induced by ristocetin. 5. ADP decomposing activity and platelet TXA2 production inhibiting activity were separated by anion exchange column chromatography of soluble BBMV. 6. When BBMV was treated with ultrasonic waves to destroy the membrane structure, only the activity inhibiting ristocetin-induced platelet aggregation was markedly decreased.

By using brush border membrane vesicles prepared from human mid gestational fetal kidney (cortex), the D-glucose transport mechanism and its developmental pattern was studied by the rapid filtration technique. 1. The transport of D-glucose into brush border membrane vesicles prepared from fetal kidney (18 gestational weeks) did not show a great degree of Na+ gradient (extravesicular greater than intravesicular) dependence. 2. The transport of D-glucose into brush border membrane vesicles prepared from fetal kidney (22 gestational weeks) depended on Na+ gradient. This Na+ dependent D-glucose transport showed overshoot and markedly increased when the intravesicular space was rendered electrically more negative with a membrane potential induced by highly permeant anions or by the K+ diffusion membrane potential via valinomycin. These results indicated that the Na+ dependent active D-glucose reabsorption system in the proximal tubules of kidney might be formed between 18 and 22 gestational weeks and the D-glucose reabsorption mechanism of kidney at 22 gestational weeks was Na+ cotransport dependent on membrane potential as in adult ones.

To investigate the placental DHA-S (dehydroepiandrosterone sulfate) transport mechanism, the uptake of DHA-S into microvillous membrane vesicles prepared from human term placenta was studied using the rapid filtration technique. 1. The uptake of DHA-S into microvillous membrane vesicles was not dependent on both Na+ electrochemical gradient and membrane potential difference. 2. The uptake of DHA-S into microvillous membrane vesicles was dependent on temperature. The initial uptake rate of DHA-S at 37 degrees C was three times as great as at 4 degrees C. 3. The initial rate of DHA-S transport exhibited saturation kinetics with respect to the DHA-S concentration; an apparent Km of 0.067 mM and Vmax of 1.01 nmol/mg protein/20 sec were calculated. 4. The uptake of DHA-S into microvillous membrane vesicles was inhibited by DHA, but not by estriol. These results indicated that placental DHA-S transport was carrier mediated and a passive one.

On the mechanism underlying the repression of antibody production encountered in the new-born, in this study we used the induction of antibody-producing cells on mitogen stimulation of PWM and SAC, identification of T-cell subsets through two-color FACS and the evaluation of cytokine production of IL-1, IL-2 and BCGF with the following results; 1. The cord blood IgG level rose with gestational weeks due to active transport from the mother. The IgM level was low in full term infants. A natural antibody first appeared in the 16th week. 2. Antibody production on mitogen stimulation in infants was remarkably less than in adults. The inability to produce antibodies appears to be closely related to T-cell factors. 3. In examination of the cell surface maker, new-born infants show less suppressor T-cell but significantly more suppressor inducer T-cell than in adults and that its much smaller number of helper T-cells are insufficient for antibody production. 4. Production of IL-1 in full term infants was significantly lower and that in premature subjects was much lower than that in adults. In full term infants, IL-2 was about the same as in adults, and BCGF production was greater than in adults not depressed in full term infants, and were significantly higher in premature infants than that in adults.

To investigate the placental L-ascorbate (Vitamin C) transport mechanism, the uptake of L-ascorbate into microvillous membrane vesicles prepared from human term placenta was studied using the rapid filtration technique. The uptake of L-ascorbate into microvillous membrane vesicles was osmotically sensitive. This finding indicated that the uptake of L-ascorbate into microvillous membrane vesicles represented transport into the vesicles. The uptake of L-ascorbate into microvillous membrane vesicles was not dependent on sodium electrochemical gradient. The initial rate of uptake was not changed when the intravesicular space was rendered electrically more negative by membrane diffusion potential induced by the use of highly permeant anions. The initial rate of L-ascorbate transport exhibited saturation kinetics with respect to the L-ascorbate concentration; an apparent Km of 1.33 mM and Vmax of 47p mol/mg protein/20 sec was calculated. The uptake of L-ascorbate into microvillous membrane vesicles was competitively inhibited by D-isoascorbate. These results indicated that transport of L-ascorbate across the placental microvillous membrane vesicles was carrier mediated and was passive transport.

Previously we reported that we succeeded in isolating from human full term placenta, glycopeptide which was immunologically the same as nephritogenic glycopeptide (Nephritogenoside) prepared from human renal GBM (glomerular basement membrane). This time to clarify the correlation between the pathology of toxemia and Nephritogenoside, we examined the existence of Nephritogenoside in the urine of toxemia patients. To purify Nephritogenoside from the urine powder of toxemia patients, we use the same procedures (enzyme digestion, Con A affinity column chromatography e.t.c.) as used in purifying Nephritogenoside from human full term placental TrBM (trophoblast basement membrane). In the urine of toxemia patients we clarified the existence of glycopeptide, which had specific affinity with Con A and was immunologically the same as Nephritogenoside. The existence of Nephritogenoside in the urine of toxemia patients further strengthened the possibility of a relationship between Nephritogenoside and the pathology of toxemia.

It is reported that ceftriaxone (Ro 13-9904, CTRX) has a half-life time of 7 to 8 hours. In the present study, the serum level 18 hours after intravenous injection with 1 g CTRX was as high as 9.3 micrograms/ml while obviously a higher tissue concentration was maintained compared with other drugs. These facts suggest that CTRX is effective against infections and that the dosage and frequency of administration could be reduced. The global evaluation revealed that CTRX was clinically effective in all the 4 cases with infections. As the adverse reaction, light leukopenia was observed only in 1 case out of the present 4 cases and 20 others administered with CTRX.

The placental transport mechanism of glutathione (GSH) was investigated using microvillous membrane vesicles prepared from human term placenta. Using (3H-glycine)-labeled-GSH, it was clarified that GSH in the extravesicular compartment of placental microvillous membranes was rapidly degraded by gamma-GTP (gamma-glutamyltranspeptidase) and resulting amino acid, and 3H-labeled-glycine was actively transported via a sodium cotransport system. AT-125 treated microvillous membrane vesicles almost entirely lost its gamma-GTP activity, and showed intact GSH transport. Using AT-125 treated microvillous membrane vesicles, it was revealed that GSH was transported across the microvillous membrane as an anion via a membrane potential-dependent mechanism. These results indicated that gamma-GTP which existed in microvillous membrane played a role in GSH metabolism and that intracellular GSH was translocated out of the syncythiotrophoblast cell into the maternal blood space via a specific carrier in microvillous membrane because the GSH concentration was higher in intracellular than extracellular and extracellular membrane potential was positively charged.

A synthetic ceramic probe allows a wider angle of irradiation and diffusion of low power energy, below 3W, using an interstitial technique for producing local hyperthermia of Nd: YAG laser (Laserthermia). We studied the influences of this laserthermia procedure in experimental animals and patients with head and neck cancer. The histological findings after laserthermia in the rabbit tongues showed no marked tissue damage in the heated area. Biochemical analysis of arachidonic acid cascade metabolites in these tissues revealed an immediate increase of 12-, 15- HETEs and PGE2 and 3 at days after hyperthermia, but at 8 days these metabolites returned to control levels. The effectiveness of laserthermia on human thyroid cancer implanted in nude mice was compared with that in cisplatin treatment and in combined therapy with both. The most predominant effectiveness was found in the animals receiving therapy combining cisplatin and laserthermia. Laserthermia in combination with radio-chemotherapy was applied in 21 cases with head and neck cancer. Good efficacy was obtained in 17 patients, especially in those with oropharyngeal cancer, showing CR in 4 cases and PR in 13 cases.

The uptake of taurocholate into microvillous membrane vesicles prepared from human normal full term placenta was studied using a rapid filtration technique. The taurocholate uptake into microvillous membrane vesicles was sensitive to extravesicular osmolarity, and preincubation with the taurocholate increased the uptake of taurocholate into the vesicles. These findings indicate that the uptake of taurocholate by microvillous membrane vesicles represents transport into vesicles. The uptake of taurocholate into vesicles was not dependent on sodium electrochemical gradient (extravesicular greater than intravesicular). But, this uptake was markedly increased when the intravesicular space was rendered electrically more positive by the use of lowly permeant anions or K+ diffusion potentials via valinomycin. These findings indicated that taurocholate was transported into microvillous membrane vesicles as anion. Cholic acid inhibited the uptake of taurocholate into vesicles, but taurine didn't inhibit this uptake. The initial rate of taurocholate transport exhibited saturation kinetics with respect to the taurocholate concentration; an apparent Km of 67 microM and Vmax of 0.30n mol/mg protein/20 sec were calculated. These results indicated that placental taurocholate transport was not active but passive (facilitated diffusion), and taurocholate was transported from fetus to mother via placenta because the blood concentration was higher in fetus than mother.

In human pregnancy, maternal and umbilical blood of the 22nd week of pregnancy and of the parturition were submitted to the analysis of total free amino acids (FAA) by amino acids analyzer. In addition, the neonatal blood was also submitted until 9 days after birth. The content of FAA in umbilical vein was higher than that of the maternal blood and it decreased at the parturition. However, the content of taurine (T) kept extremely higher level than other amino acids until term and it decreased to the normal level at 9 days after birth. In pregnant rats on day 17, 18 and 19 of gestation, the content of FAA in umbilical vein and fetal artery were 5800-7000 micrograms/dl and 5700-6250 micrograms/dl, respectively. They were two fold higher than that of maternal value. In amniotic fluid, FAA was 4100 micrograms/dl on day 18 and 8850 micrograms/dl on day 20. On the contrary, the content of T in maternal blood on day 18 was the highest value (360 micrograms /dl), and decreased to the half of the value on day 20. T in carotid artery on the same day was 1150 micrograms/dl, but it decreased to 280 micrograms/dl on day 20. In amniotic fluid, it was almost constant in the range of 380-400 micrograms/dl. When the pregnant rat was stressed by starvation, FAA was increased in both umbilical and fetal blood but it was almost constant in amniotic fluid. T was increased both in maternal and fetal blood. When the degree of starvation, T value also increased in fetal blood. From these data, we discussed on the possible mechanism of the production of T during pregnancy and after birth.

A 27-year-old woman was admitted to our hospital complaining of purpuras and legs' edema in the 38th week of pregnancy. On admission, the hemoglobin was 7.8 g/dl, platelets 20,000/microliter and WBC 6,600/microliters with 52% blast cells. Bone marrow aspirate demonstrated 77.2% myeloblasts with prominent Auer rods, consistent with acute myelogenous leukemia. Receiving packed-red-cell and platelet transfusions, she delivered a normal male infant in the 39th week of pregnancy by normal labor. After delivery, she was placed on a combination chemotherapy of BHAC-MMP, subsequently DNR added. Four weeks later, a complete remission was obtained, lasting for almost one year, and her child has grown well without hematological disorder. Microscopic findings of the placenta obtained at delivery revealed no invasion of leukemic cells, but 9% blast cells were present in the placental cord blood. We reviewed 18 cases reported in Japan of acute leukemia in gestational period, that could obtain complete remission and keep the children growing well. Placental transmission of leukemic cells from mother to infant was discussed.

To study the toxic effect of inorganic mercury on the placenta we elucidate the effect of inorganic mercury on placental amino acid transport using microvillous membrane vesicles isolated from human normal full term placenta. The transport of amino acids into microvillous membrane vesicles was studied by a rapid filtration technique using a millipore filter. The transport of L-alanine across placental microvillous membrane was Na+ electrochemical gradient dependent and 0.1 mM inorganic mercury inhibited 77% of this Na+ dependent L-alanine transport and 1 mM inorganic mercury inhibits 90% of this Na+ dependent L-alanine transport. The transport of L-lysine across microvillous membrane vesicles was sodium independent and 0.1 mM inorganic mercury inhibited 34% of this transport and 1 mM inorganic mercury inhibited 50% of this transport. These results indicated that one of the toxic effects of inorganic mercury on placenta-fetus unit was the inhibition of placental nutrient transport.

In order to elucidate the effect of antibiotics (gentamicin) on placental amino acids transport, we investigated L-alanine transport using microvillous membrane vesicles prepared from full-term human placental by a rapid filtration technique. 1. The active transport of L-alanine into microvillous membrane vesicles was dependent on Na+ electrochemical gradient (extravesicular greater than intravesicular). And the double reciprocal plot of this Na+ dependent initial uptake rate versus L-alanine concentration exhibited an apparent Km of 0.79 + 0.23mM and a Vmax of 3.56 + 0.70n mol/mg protein/20 sec. 2. Gentamicin did not affect the Km value of this Na+ dependent L-alanine transport kinetics (0.77 + 0.19 mM [lmM gentamicin], 0.79 + 0.21mM [10mM gentamicin]). On the other hand, gentamicin apparently decreased, the Vmax value of this transport kinetics (1.99 + 0.48n mol/mg protein/20 sec [1mM gentamicin], 1.12 + 0.32n mol/mg protein/20 sec [10mM gentamicin]).

Previously, we reported that we succeeded in isolating from human placenta a substance comparable to human renal nephritogenic glycoprotein prepared from glomerular basement membrane (GBM). This time we purified a sample isolated from human placental trophoblast basement membrane (TrBM) with the methods for purifying renal nephritogenic glycoprotein and clarified its chemical composition. To purify the sample prepared from human placental TrBM, we fractionated the sample by Zone Electrophoresis, Concanavalin A(Con A) affinity column chromatography and Bio Gel P200 column chromatography. As the active fraction of the sample purified from human placental TrBM bound specifically with Con A, it proved to be a glycoprotein. The monosaccharide composition of this glycoprotein was rich in glucose (glucose, galactose and mannose were in the ratio of 1.00:0.30:0.33) and the amino acid composition of this glycoprotein contained no collagenous components. Ouchterlony gel diffusion demonstrated the existence of a common precipitin line between antiserum to human renal nephritogenic glycoprotein prepared from GBM and human renal nephritogenic glycoprotein prepared from GBM, purified human placental glycoprotein, the sample prepared from human placental TrBM by sonication and nephritogenic glycoprotein prepared from urine of S.L.E. patient.

To elucidate the change in EPH gestosis placental amino acid transport activity, we investigated the uptake of L-alanine into microvillous membrane vesicles prepared from EPH gestosis placenta and from normal placenta by using a rapid filtration technique. 1. Alkaline phosphatase (ALP) was the marker enzyme of microvillous membrane vesicles (MMV). The ALP activity of mild EPH gestosis placental MMV didn't differ from that of normal placental MMV. On the other hand, the ALP activity of severe EPH gestosis placental MMV decreased compared to that of normal placental MMV. 2. The uptake of L-alanine into human placental MMV was dependent on the Na+ electrochemical gradient, so the transport across human placental MMV was a secondarily active one. The L-alanine transport activity of mild EPH gestosis placental MMV didn't differ from that of normal placental MMV. On the other hand, the L-alanine transport activity of severe EPH gestosis placental MMV decreased prominently compared to that of normal placental MMV.

The uptake of L-lysine in human placental microvilli vesicles prepared from human term placenta was studied using the rapid filtration technique. The uptake of L-lysine into the vesicles was osmotically sensitive. This finding indicates that the uptake of L-lysine by placental microvilli vesicles represents transport into the membrane vesicles. The uptake of L-lysine into microvilli vesicles is sodium independent. The initial rate of uptake is markedly increased when the intravesicular space is rendered electrically more negative by membrane diffusion potential induced by the use of highly permeant anions or by K+ diffusion potentials via valinomycin. These results indicate that the sodium independent uptake of L-lysine into the microvilli membrane vesicles is dependent on the electrical potential difference of the membranes. A kinetic analysis of the uptake demonstrated that two transport systems for vesicular entry of L-lysine existed with a Km1 of 0.13 mM, Vmax1 of 590 pmol/mg protein/20 sec, Km2 of 0.91 mM, Vmax2 of 2010 pmol/mg protein/20 sec. The uptake of L-lysine in microvilli vesicles was inhibited by dibasic amino acid (L-arginine, L-ornithine, L-glutamine), but not by other classes of amino acid. These results indicate the existence of a dibasic amino acid specific transport system in placental microvilli membrane.

By using brush border membrane vesicles prepared from human mid gestational fetal intestine (jejunum), the intestinal L-alanine transport mechanism was studied using a rapid filtration technique. The uptake of L-alanine by the vesicles was osmotically sensitive. This finding indicated that the uptake of L-alanine by the vesicles represented transport into the vesicles. A Na+ electrochemical gradient (extravesicular greater than intravesicular) stimulated the initial rate of L-alanine uptake, and Na+ dependent uptake of L-alanine into vesicles showed a typical overshoot phenomenon. This overshoot and the initial rate of uptake markedly increased when the intravesicular space was rendered electrically more negative by membrane diffusion potentials induced by with highly permeant anions. A similar stimulation of L-alanine uptake was observed, when membrane potential (inside negative) was imposed by K+ diffusion potentials via valinomycin. These results indicated that an Na+ dependent uptake of L-alanine into the brush border membrane vesicles was dependent on the electrical potential difference of the membrane. The initial rate of L-alanine transport exhibited saturation kinetics with respect to the L-alanine concentration; an apparent Km of Km1; 1.6mM, Km2; 14.1mM, and Vmax of Vmax1; 4.0n mol/mg protein/20sec, Vmax2; 8.2n mol/mg protein/20 sec were calculated. From these results mid gestational fetal intestine (jejunum) already has an L-alanine absorption system which is comparable to the adult one's.

To elucidate the human placental L-lactate transport mechanism, we investigated L-lactate uptake by microvillous membrane vesicles (MMV) prepared from full termed human placenta using the rapid filtration technique. 1. The transport of L-lactate into the MMV was not dependent on the Na+ electrochemical gradient (extravesicular greater than intravesicular). 2. The transport of L-lactate into the MMV was dependent on the H+ gradient (extravesicular greater than intravesicular) and this H+ dependent L-lactate uptake showed a tendency to overshoot. This overshoot disappeared with the addition of H+ ionophore (FCCP). 3. The initial rate of this H+ dependent L-lactate transport into the MMV exhibited saturation kinetics with respect to the L-lactate concentration: An apparent Km of 4.35mM and Vmax of 2.78n mol/mg protein/20sec were calculated. These results indicated that there existed a H+ and L-lactate transport system in human placental MMV.

Sulbactam/cefoperazone (SBT/CPZ), a new developed antibiotic, was clinically studied. The following results were obtained. Total of 5 cases comprising 1 with endometritis, 3 with Bartholin's abscess, 1 with pyometra (due to corpus cancer) were treated with SBT/CPZ at a dose of 1 g twice daily for 4-5 days by intravenous injection or intravenous drip infusion. The clinical response was good in all cases. A case with pyometra did not respond to the therapy in bacteriologically. Side effects and abnormal laboratory findings due to the drug were not noted.

The effect of potassium ion and membrane potential on the uptake of L-glutamate in microvilli (brush border) vesicles prepared from human term placenta was studied using a rapid filtration technique. The uptake of L-glutamate into microvilli vesicles is Na+ electrochemical gradient dependent and pre-equilibration of the vesicles with K+ stimulates L-glutamic acid uptake. Imposition of a K+ gradient (K+ in greater than K+ out) enhances Na+-dependent L-glutamate uptake. Changes in membrane potential due to the imposition of anion replacement markedly affect Na+ dependent L-glutamate uptake only in the presence of K+. However, this effect is not significant when changes in membrane potential incur following the imposition of valinomycin induced by K+ diffusion potential. The data indicate that Na+-dependent L-glutamate transport can be additionally energized by a K+ gradient. Furthermore K+ renders Na+-dependent L-glutamate transport sensitive to changes in the transmembrane potential difference.

Using microvillous (brush border) membrane vesicles prepared from human term placenta, the uptake of L-glutamine was studied using a rapid filtration technique. The uptake of L-glutamine into the vesicles was osmotically sensitive. A Na+ electrochemical gradient (extravesicular greater than intravesicular) stimulated the initial rate of L-glutamine uptake and the Na+ dependent uptake of L-glutamine into vesicles showed a typical overshoot phenomenon. This overshoot and the initial rate of uptake were markedly increased when the intravesicular space was rendered electrically more negative by membrane diffusion potentials, induced by the use of highly permeant anions. A similar stimulation of L-glutamine uptake was observed when membrane potential (inside negative) was imposed by K+ diffusion potentials via valinomycin. These results indicated that a sodium dependent uptake of L-glutamine into the microvillous membrane vesicles was dependent on the electrical potential difference of membrane. The initial rate of L-glutamine transport exhibited saturation kinetics with respect to L-glutamine concentration; the apparent Km of 0.42 mM and Vmax of 1.54 nmol/mg protein/20 sec were calculated. The uptake of L-glutamine into the vesicles was competitively inhibited by L-alanine.

To elucidate the character of human placental D-glucose and L-alanine transport activity, we investigated the uptake of D-glucose and L-alanine into microvillous membrane vesicles prepared from human early and term placenta using the rapid filtration technique. 1. The uptake of D-glucose into microvillous membrane vesicles did not depend on the Na+ gradient (extravesicular greater than intravesicular). The uptake of D-glucose into vesicles was three times as great as that of L-glucose. And phloretin prominently inhibited the uptake of D-glucose into vesicles. So, it was indicated that the transport mechanism of D-glucose across microvillous membrane was facilitated diffusion. 2. The uptake of L-alanine into microvillous membrane vesicles depended on Na+ gradient (extravesicular greater than intravesicular), so that the transport mechanism of L-alanine across microvillous membrane was a secondary active one. The transport of L-alanine into vesicles prepared from term placenta increased prominently compared to that of early placenta. On the other hand, the transport activity of D-glucose into vesicles prepared from term placenta did not differ from that of early placenta.

Breast-feeding is a major factor in the vertical transmission of human T-lymphotropic virus type I (HTLV-I). We studied whether such transmission may be prevented by bottle-feeding. HTLV-I infection was detected by both HTLV-I antigen and antibody tests. Thirty bottle-fed babies were examined 24 months after birth; only one was found to be HTLV-I antigen-positive. This infection rate was lower than that for breast-fed babies in whom HTLV-I antigen was detected in 24 of the 31 24-month-old babies born to HTLV-I positive mothers in a previous study. These results suggest that most vertical transmission of HTLV-I is attributable to breast-feeding and can be prevented by bottle-feeding.

Cefodizime (THR-221, CDZM), a new antibiotic, was studied pharmacokinetically and clinically. The results obtained are summarized as follows: 1. Concentrations of CDZM in internal genital tissues were quite high after an intravenous infusion. 2. Clinical effects of the therapy with CDZM using intravenous infusion twice daily were evaluated in 1 patient with endometritis, 2 patients with pyometra, 1 patient with extragenital abscess and 1 patient with BARTHOLIN'S gland abscess. Clinical responses were good in all 5 patients. No side effects nor abnormal laboratory test values due to the drug were noted.