Synthesis of 4-Imidazolidinones from Diamides and Ethynyl Benziodoxolones via Double Michael-Type Addition: Ethynyl Benziodoxolones as Electrophilic Ynol Synthons.

The moiety of 4-imidazolidinone is an important structural motif in organic synthesis and medicinal chemistry. We present the synthesis of 4-imidazolidinones from various diamides with ethynyl benziodoxolones through double Michael-type addition, which is an unprecedented reaction mode for hypervalent alkynyl iodine compounds. cis-2,5-Disubstituted 4-imidazolidinones were diastereoselectively synthesized from amino acid derived diamides. Having derivatized the 4-imidazolidinones, several control experiments and density functional theory calculations were conducted to realize mechanistic insight.

Triboelectric charging of polytetrafluoroethylene antithrombotic catheters.

This study proposes that a polytetrafluoroethylene (PTFE) electret tube charged by frictional electricity can prevent the solidification of the indwelling catheter in blood vessels. Coagulation in intravascular indwelling catheters may discontinue the treatment because of thrombus-derived bacteria-adhesion infections or poor blood removal. Current commercially available intravascular catheters lack complete antithrombotic measures, even with heparin or urokinase antithrombotic coatings. Herein, we tested the effectiveness of an antithrombotic treatment that prevents coagulation using a static electric charge on the interior of the PTFE tube via the triboelectric effect by rubbing the tube's inner wall with a round glass rod. The anticoagulation properties were evaluated by enclosing a sample of blood in an electret tube and observing the coagulase adhering to the inner wall using a microscope. To confirm the effectiveness of this treatment, the charge-distribution on the inner surface of the electret tube was measured, surface irregularities were observed, and the elements on the surface were analyzed. The surface potential inside the electret tube was - 366.4 V, which proved effective for an antithrombotic treatment, as it discouraged coagulation, and the triboelectric charging process caused neither surface element denaturation nor significant surface irregularities. The nearly uniform negative surface charge on the inside of the tube was responsible for the antithrombotic effect because no surface irregularities or change in the surface element denaturation was observed. Triboelectrically charged PTFE electret tubes are highly useful for intravascular indwelling catheters.

Contribution of radixin to P-glycoprotein expression and transport activity in mouse small intestine in vivo.

The ERM proteins, ezrin, radixin, and moesin, are membrane-cytoskeleton cross-linkers with multiple physiological functions. We previously showed that radixin is involved in posttranslational regulation of P-glycoprotein (P-gp) in human hepatoblastoma HepG2 cells. Here, we investigated the physiological role of radixin in regulating P-gp expression and activity in the small intestine by comparing wild-type- and radixin knockout (Rdx) mice. In intestinal tissue homogenates, P-gp protein levels increased markedly from the upper part to the lower part of the small intestine in both wild-type- and Rdx(-/-) mice. In the membrane fractions, a similar pattern was seen in wild-type mice. However, the membrane expression of P-gp protein remained at the same level from the upper to the lower part of the small intestine in Rdx(-/-) mice. When rhodamine123 (Rho123), a substrate of P-gp, was orally administered to Rdx(-/-) and wild-type mice, the absorption phase of Rho123 was greater in Rdx(-/-) than in wild-type mice, whereas the elimination phase in Rdx(-/-) mice was not different from that of wild-type mice. Our results indicate that radixin plays an important role in regulating P-gp localization and P-gp functional activity at the intestinal membrane.

The role of inter-segmental differences in P-glycoprotein expression and activity along the rat small intestine in causing the double-peak phenomenon of substrate plasma concentration.

  Conflicting results have been reported on segmental differences in expression of P-glycoprotein (P-gp) along the small intestine of animals and humans. In this study, we investigated P-gp mRNA and protein levels within each of nine segments of rat small intestine. In addition, P-gp activity in each segment was evaluated in terms of permeability of rhodamine123 (Rho123), a typical P-gp substrate, using the serial intestinal non-everted sac method. The P-gp mRNA levels tended to increase from the duodenum to the ileum, with peaks in the upper and lower ileum, while P-gp protein level reached its maximum in the middle ileum. The activity of P-gp was also the highest in the middle ileum, and was highly correlated with P-gp protein level. The double-peaked plasma concentration profile that was observed following oral administration of Rho123 to rats could be well reproduced by an intestinal compartmental kinetic model incorporating inter-segmental differences of absorption and excretion rate constants. Our results suggest that the heterogeneous distribution of P-gp along the small intestine plays a key role in causing the double-peak of plasma concentration of P-gp substrates following oral administration to rats.

Developmental changes of brain distribution and localization of oseltamivir and its active metabolite Ro 64-0802 in rats.

Oseltamivir, a prodrug of the neuraminidase inhibitor [3R, 4R, 5S]-4-Acetamide-5-amino-3-(1-ethylpropyl)-1-cyclohexene-1-carboxylate phosphate (Ro 64-0802), is widely used for treatment of influenza infections in Japan, but may be associated with mental instability and suicidal tendencies as a rare side effect, especially in infants and young patients. We examined developmental changes in the brain distribution of oseltamivir and Ro 64-0802, and in the expression of P-glycoprotein (P-gp) at the blood-brain barrier (BBB) in rats by 8 weeks. Brain concentration and Kp(,app,brain) (brain-to-plasma concentration ratio) of oseltamivir were highest in 2-week-old rats (1.45 µg/g brain and 0.14, respectively), and were negatively correlated with both age and P-gp expression at the BBB. In contrast, brain concentration and Kp(,app,brain) of Ro 64-0802 after oral gavage of oseltamivir were lowest in 2-week-old rats (0.02 µg/g brain and 0.02), and increased with age. Mass imaging analysis revealed that both compounds were distributed homogenously in brain cross-sections, including the hippocampus. From these results, it was estimated that oseltamivir concentration throughout the brain cross-sections was 70-fold and 0.9-fold higher than that of Ro 64-0802 in 2-week-old and 8-week-old rats, respectively. Such developmental changes of prodrug/drug concentration ratio, if they also occur in humans, may provide a rational basis for the putative central nervous system (CNS) side effects in young patients.

Effect of knockdown of ezrin, radixin, and moesin on P-glycoprotein function in HepG2 cells.

Ezrin, radixin, and moesin (ERM) proteins regulate functional expression of certain transporters, but little is known about their effect on P-glycoprotein (P-gp). Here, we investigated the influence of ERM proteins on the expression and activity of P-gp at the transcriptional, translational, and posttranslational levels, using HepG2 as a model cell line. Knockdown of ezrin with RNA interference decreased the level of P-gp messenger RNA. On the contrary, knockdown of radixin caused a decrease of the P-gp gene product at the cell surface, but not in whole cell lysate. Furthermore, a significant increase in accumulation of rhodamine123, a typical P-gp substrate, was observed in radixin knockdown cells, compared with control cells. Knockdown of moesin did not influence the expression or function of P-gp. These results indicate that ezrin influences the expression of P-gp at the translational level, whereas radixin is involved in membrane localization of P-gp in HepG2 cells.

Effect of milk on the pharmacokinetics of oseltamivir in healthy volunteers.

We previously showed that oseltamivir, a prodrug of the influenza virus neuraminidase inhibitor Ro 64-0802, is a substrate of proton-coupled oligopeptide transporter (PEPT1), and its intestinal absorption in rats is markedly inhibited by administration with milk. To investigate the importance of PEPT1 for oseltamivir absorption in humans, and the characteristics of the drug-milk interaction, a crossover clinical study was conducted in healthy volunteers, who received 75 mg of oseltamivir with 400 mL of water or milk. Milk significantly reduced the maximum plasma concentration (C(max) ) and the area under the plasma concentration-time curve from 0 to 2 h (AUC(0-2) ) of both oseltamivir and Ro 64-0802 (oseltamivir, 68.9% and 34.5%; Ro 64-0802, 69.5% and 14.2%, respectively, vs. water), but had no significant effect on the apparent terminal half-life (t(1/2) ) or AUC(0-∞) . Urinary recovery of oseltamivir and Ro 64-0802 was significantly reduced to 77.5% of the control by milk. The early reduction of oseltamivir absorption might be through the PEPT1 inhibition by milk peptides. However, the extent of interaction in humans was limited as compared with that in rats, possibly because of species difference in the PEPT1 expression and its contribution. This might be the first report suggesting the clinical drug-food interaction via PEPT1.

Enhancement of drug solubility and absorption by copolymers of 2-methacryloyloxyethyl phosphorylcholine and n-butyl methacrylate.

Poly[2-methacryloyloxyethyl phosphorylcholine-co-n-butyl methacrylate]s (PMBs) are water-soluble solid copolymers of 2-methacryloyloxyethyl phosphorylcholine (MPC) and n-butyl methacrylate with a molecular weight of 30,000 (PMB50T) or 100,000 (PMB100T). Here, we characterized the solubilizing properties of PMBs using miconazole (MCZ), vidarabine (Ara-A) and griseofulvin (GRF), which are class 2, 3 and 4 compounds, respectively, in the Biopharmaceutics Classification System (BCS). Moreover, we evaluated the enhancement of gastric absorption of GRF dissolved in PMB solutions and the toxicity of PMBs in rats. PMB50T solution dramatically increased the solubility of GRF and MCZ compared with Ara-A, and these drugs became more soluble as the concentration of PMB50T was increased. The solubility of GRF in 10% PMB solutions was higher than with any other tested aqueous solubilizer. When a solution of GRF (20 mg/10 mL/kg) in 10% PMB was orally administered to rats, GRF absorption was greatly increased compared with that following administration of a suspension in water or Gelucire. After repeated oral administration of PMBs once daily for 14 successive days, no organ lesions or changes in biochemical parameters were observed. Thus, the polymers are expected to be useful and safe solubilizers and oral absorption enhancers for poorly soluble lipophilic drugs.

Sairei-to therapy on alloimmune recurrent spontaneous abortions and alloimmune-, autoimmune complicated recurrent spontaneous abortions.

Alloimmune recurrent spontaneous abortion (RSA) cases that could not be treated with lymphocyte transfusion due to medical and social reasons were treated with Sairei-to therapy as an emergency measure and all four cases resulted in live births. This may show that Sairei-to treatment is effective in preventing alloimmune RSA. The efficacy of Sojyutsu-Sairei-to and Byakujyutsu-Sairei-to on autoimmune RSA has already been proven. When they were used in the treatment of alloimmune-, autoimmune complicated RSA, the abortion prevention rates were 65.4% and 82.3% respectively. These results indicate that Sairei-to is effective in the treatment of alloimmune RSA and alloimmune-, autoimmune complicated RSA.

Differences in individual efficacy of two Sairei-to preparations (Sojyutu-Sairei-to and Byakujyutu-Sairei-to) on recurrent spontaneous abortions of autoimmune etiologies evaluated by antinuclear antibody and anticardiolipin antibody titers.

The differences in individual efficacy of two Sairei-to preparations (Sojyutu-Sairei-to and Byakujyutu-Sairei-to) on antinuclear antibody (ANA) and anticardiolipin antibody (ACLA) positive recurrent spontaneous abortion (RSA) was analyzed in 52 patients (a total of 61 treatment sessions). Patients who failed to respond to initial treatment with Sojyutu-Sairei-to were additionally treated with Byakujyutu- Sairei-to, and the time course of ANA and ACLA titers in these patients was analyzed. ACLA titers were decreased significantly by the treatment of Byakujyutu-Sairei-to, however, the percentage of successfully prevented abortion cases did not differ significantly between the Sojyutu-Sairei-to treatment group and the Byakujyutu-Sairei-to treatment group. ACLA titer was decreased in all 10 cases where abortion was successfully prevented by the treatment with Sojyutu-Sairei-to or Byakujyutu-Sairei-to. In the cases where both ANA and ACLA were decreased following treatment with Sojyutu-Sairei-to or Byakujyutu-Sairei-to, the percentage of cases rated as "Kyo" and "Rikan" were significantly higher in the Byakujyutu-Sairei-to group. These results indicate that Byakujyutu-Sairei-to is effective against ACLA positive RSA through the antibody-reducing activity, which differs from that of Sojyutu-Sairei-to in individual cases. On the basis of these results, Sairei-to therapy, which is superior to aspirin and heparin in terms of efficacy and safety, is recommended as the first-line therapy for RSA of autoimmune etiologies. Furthermore, to elevate the percentage of successfully prevented abortions, it is advisable to select one of the two Sairei-to preparations (Sojyutu-Sairei-to and Byakujyutu-Sairei-to) on the basis of differential diagnosis using the methods of Oriental medicine.

Gender ratio distortion in abortuses and live births from patients with recurrent spontaneous abortion.

PROBLEM: Gender ratio of live birth in humans is approximately 1.05 and males are born a slightly more, while gender ratio of fertilization should be 1.00, suggesting that female fetus might be more sensitive to abortion than male fetus during pregnancy. METHOD OF STUDY: We examined karyotype of abortuses from patients with recurrent spontaneous abortion (RSA), who had at least one live birth before or after the treatment of RSA. RESULTS: Chromosomal abnormality was not frequent (14.6%) in the abortuses from the RSA patients. Among abortuses without chromosomal abnormality, male karyotype was rare (9.2%), and this gender ratio distortion was more prominent in RSA cases not carrying autoantibodies (3.5%) than that in the RSA cases carrying autoantibodies (26.3%), with statistical significance (P = 0.009). CONCLUSION: These observations suggested that the aborted fetuses from RSA of unknown etiology, i.e. no chromosomal abnormality and no autoantibody, were preferentially female.

Evaluation of the inhibitory effect of dihydropyridines on N-type calcium channel by virtual three-dimensional pharmacophore modeling.

Currently, a new type of calcium channel blockers, which can inhibit not only L-type calcium channels abundantly expressed in vascular smooth muscles, but also N-type calcium channels that abound in the sympathetic nerve endings, have been developed. In this study, analysis on a like-for-like basis of the L- and N-type calcium channel-inhibitory activity of typical dihydropyridine-type calcium-channel blockers (DHPs) was performed. Moreover, to understand the differences of N-type calcium channel inhibition among DHPs, the binding of DHPs to the channel was investigated by means of hypothetical three-dimensional pharmacophore modeling using multiple calculated low-energy conformers of the DHPs. All of the tested compounds, i.e. cilnidipine (CAS 132203-70-4), efonidipine (CAS 111011-76-8), amlodipine (CAS 111470-99-6), benidipine (CAS 85387-35-5), azelnidipine (CAS 123524-52-7) and nifedipine (CAS 21829-25-4), potently inhibited the L-type calcium channel, whereas only cilnidipine inhibited the N-type calcium channel (IC50 value: 51.2 nM). A virtual three-dimensional structure of the N-type calcium channel was generated by using the structure of the peptide omega-conotoxin GVIA, a standard inhibitor of the channel, and cilnidipine was found to fit well into this pharmacophore model. Lipophilic potential maps of omega-conotoxin GVIA and cilnidipine supported this finding. Conformational overlay of cilnidipine and the other DHPs indicated that amlodipine and nifedipine were not compatible with the pharmacophore model because they did not contain an aromatic ring that was functionally equivalent to Tyr13 of omega-conotoxin GVIA. Azelnidipine, benidipine, and efonidipine, which have this type of aromatic ring, were not positively identified due to intrusions into the excluded volume. Estimation of virtual three-dimensional structures of proteins, such as ion channels, by using standard substrates and/or inhibitors may be a useful method to explore the mechanisms of pharmacological and toxicological effects of substrates and/or inhibitors, and to discover new drugs.

Oseltamivir (tamiflu) is a substrate of peptide transporter 1.

Oseltamivir, an ester-type prodrug of the neuraminidase inhibitor [3R,4R,5S]-4-acetamido-5-amino-3-(1-ethylpropoxy)-1-cyclohexene-1-carboxylate phosphate (Ro 64-0802), has been developed for the treatment of A and B strains of the influenza virus but has neuropsychiatric and other side effects. In this study, we characterized the transport across intestinal epithelial cells and the absorption of oseltamivir in rats. Uptake by Caco-2 cells (human carcinoma cell line) and HeLa cells transfected with peptide transporter 1 (HeLa/PEPT1) was time- and temperature-dependent and was inhibited by typical PEPT1 inhibitors such as glycyl-sarcosine (Gly-Sar). The uptake by Caco-2 cells and HeLa/PEPT1 was saturable, with similar K(m) values. Oseltamivir absorption in adult rats was greatly reduced by simultaneous administration of milk, casein, or Gly-Sar. Furthermore, the plasma and brain concentrations of oseltamivir were higher in fasting than in nonfasting rats after oral administration. These results suggest that oseltamivir is a substrate of PEPT1 and that PEPT1 is involved in its intestinal absorption.

Carnitine/organic cation transporter OCTN2 (Slc22a5) is responsible for renal secretion of cephaloridine in mice.

Carnitine/organic cation transporter (OCTN) 2 (SLC22A5) plays a pivotal role in renal tubular reabsorption of carnitine, a vitamin-like compound, on apical membranes of proximal tubules, but its role in relation to therapeutic drugs remains to be clarified. The purpose of the present study was to elucidate the involvement of OCTN2 in renal disposition of a beta-lactam antibiotic, cephaloridine (CER), based on experiments with juvenile visceral steatosis (jvs) mice, which have a functional deficiency of the octn2 gene. Renal clearance of CER during constant intravenous infusion in wild-type mice was much higher than could be accounted for by glomerular filtration, but was decreased by increasing the infusion rate with minimal change in kidney-to-plasma concentration ratio, suggesting the existence of saturable transport mechanism(s) across the apical membranes. The plasma concentration profile and kidney-to-plasma concentration ratio after intravenous injection in jvs mice were higher than those in wild-type mice, whereas renal clearance in jvs mice was much lower than that in wild-type mice and could be accounted for by glomerular filtration. Uptake of CER by mouse OCTN2 was shown in Xenopus laevis oocytes expressing mouse OCTN2. The CER transport by OCTN2 exhibited saturation with K(m) of approximately 3 mM, which is similar to the renal CER concentration exhibiting saturation in renal clearance in vivo. The OCTN2-mediated CER transport was inhibited by carnitine and independent of Na(+) replacement in the medium. These results show OCTN2 on apical membranes of proximal tubules plays a major role in renal secretion of CER in mice.

Involvement of influx and efflux transport systems in gastrointestinal absorption of celiprolol.

Gastrointestinal absorption of several beta-blockers is inhibited by citrus juices, although molecular mechanism(s) lying on their small intestinal absorption has not yet been identified. Here, we attempted to demonstrate involvement of both influx and efflux transporters in vivo in gastrointestinal absorption of celiprolol in mice. Plasma concentration of celiprolol (3 mg/kg) after oral administration was mostly under the limit of quantification in wild mice, whereas that in mdr1a/b knockout (mdr1a/b(-/-)) mice was much more obvious, indicating P-glycoprotein-mediated efflux. Then, the oral absorption of celiprolol in mdr1a/b(-/-) mice was further examined to investigate influx transport mechanism with avoiding effect of P-glycoprotein. Coadministration of bromosulfophthalein (BSP), an inhibitor of various influx transporters including organic anion transporting polypeptide (OATP) reduced plasma celiprolol concentration. Inhibition by BSP of celiprolol uptake from apical membranes was confirmed in Ussing-type chamber of small intestinal tissues. Uptake of celiprolol by human small intestinal transporter OATP-A/1A2 was also confirmed in Xenopus Laevis oocytes. Interestingly, OATP-A/1A2 accepts various beta-blockers including acebutolol, atenolol and sotalol, oral absorption of which is inhibited by coadministration of citrus juice or telithromycin in human. Taken together, these findings have suggested fundamental role of influx transport system(s) in oral absorption of celiprolol.

Treatment for intractable anemia with the traditional Chinese medicines Hominis Placenta and Cervi Cornus Colla (deer antler glue).

OBJECTIVE: Intractable anemia, such as aplastic anemia or that presumably associated with chronic herpes virus infections, sometimes require bone marrow transplant. We investigated the use of traditional Chinese medicine (TCM) for the treatment of intractable anemia. METHOD: Placenta Hominis (PH), steam boiled and roasted, and Cervi Cornus Colla (deer antler glue) has been used in China for hundreds of years to treat anemia. After consent was obtained, we prescribed these two materials for a 74-year-old female with aplastic anemia and a 26-year-old male with presumably a virus-induced anemia. Concomitant conventional therapy was continued in both patients as prescribed by their respective attending physicians. CONCLUSION: Conventional therapy with steroid hormones, immunosuppressive drugs, platelet and erythrocyte transfusions were not effective in these patients. In addition, both patients suffered from serious side effects. In two patients, ingestion of Placenta Hominis and Cervi Cornus Colla with TCM prescriptions increased the platelet and enhanced the hemoglobin concentration in several months of therapy accompanied by a dramatic improvement in quality of life. The addition to conventional therapy of PH and Cervi Cornus Colla, the latter of which is very easy to obtain, may be one of the potentially advantageous choices in case of otherwise intractable anemia.

Kampo medicine, based on traditional medicine theory, in treating uncured glossodynia: efficacy in five clinical cases.

Glossodynia, or tongue pain, is resistant to conventional therapies. Kampo medicines were evaluated in patients suffering from incurable glossodynia. Patients were diagnosed by traditional Chinese medicine (TCM) theory in order to determine the appropriate herbal prescriptions. Five Japanese females (50-76 years old) with glossodynia refractory to conventional therapy were enrolled in this study. Small portions of rikkunshito, jiinkokato, hachimijiogan and ryutanshakanto worked for a female diagnosed with "Spleen" and "Heart" Yin deficiency, "Kidney" Yang deficiency and "Liver" Qi stagnation producing heat syndrome. Seishoekkito and bakumondoto were effective for a patient diagnosed with "Spleen Qi" deficiency and "Stomach" Yin deficiency producing heat syndrome. Rikkunshito, kamikihito and chikujountanto worked for a patient diagnosed with "Spleen Qi" and "Heart Yin" deficiency, stagnation of "Liver" Qi producing fire and "Gallbladder" Qi deficiency. Rokumijiogan, kamishoyosan and kambakutaisoto were effective for a patient with Yang rise based on Yin deficiency of "Kidney" and "Liver," and restless organ disorder based on Yin deficiency of 5 viscera. A patient diagnosed with "Spleen" Yang deficiency responded to a combination of anchusan and hangeshashinto. These patients with glossodynia had resolution of pain within 1 month of treatment. Herbal mixtures containing Ganoderam lucidum, not prescribed based on TCM theory, but effective for herpes virus infection, worked for a female suffering from glossodynia for 1 year after artificial teeth were placed, but required about 5 months to note improvement. Kampo medicines, properly prescribed based on TCM theory, quickly resolved the pain of refractory glossodynia.

Memory in Caenorhabditis elegans is mediated by NMDA-type ionotropic glutamate receptors.

Learning and memory are essential processes of both vertebrate and invertebrate nervous systems that allow animals to survive and reproduce. The neurotransmitter glutamate signals via ionotropic glutamate receptors (iGluRs) that have been linked to learning and memory formation; however, the signaling pathways that contribute to these behaviors are still not well understood. We therefore undertook a genetic and electrophysiological analysis of learning and memory in the nematode Caenorhabditis elegans. Here, we show that two genes, nmr-1 and nmr-2, are predicted to encode the subunits of an NMDA-type (NMDAR) iGluR that is necessary for memory retention in C. elegans. We cloned nmr-2, generated a deletion mutation in the gene, and showed that like nmr-1, nmr-2 is required for in vivo NMDA-gated currents. Using an associative-learning paradigm that pairs starvation with the attractant NaCl, we also showed that the memory of a learned avoidance response is dependent on NMR-1 and NMR-2 and that expression of NMDARs in a single pair of interneurons is sufficient for normal memory. Our results provide new insights into the molecular and cellular mechanisms underlying the memory of a learned event.

Human leukocyte antigen may predict outcome of primary recurrent spontaneous abortion treated with paternal lymphocyte alloimmunization therapy.

PROBLEM: Recurrent spontaneous abortion (RSA) is defined by at least three consecutive abortions in otherwise healthy couples. Paternal lymphocyte alloimmunization therapy (PLAT) is an effective therapy for RSA in some cases, but there are no predictive markers about the effectiveness of PLAT. METHOD OF STUDY: Forty-two consecutive cases with primary RSA treated by PLAT and 23 controls were the subjects. Polymorphisms of human leukocyte antigen (HLA)-E, HLA-G, HLA-A, HLA-B, HLA-C and HLA-DRB1 were investigated by sequenced based typing. Promoter polymorphism and a 14 bp ins/del polymorphism in exon 8 were also investigated for HLA-G. RESULTS: Thirty-eight RSA wives became pregnant within 1 year after PLAT. Among them, 27 obtained babies (succeeded PLAT cases), while 11 again aborted with no detectable chromosomal abnormalities in the aborted fetuses (aborted PLAT cases). The frequencies of HLA-G*010401, A*2402, B*5201, and DRB1*1502 were significantly increased in the aborted cases than those in the succeeded cases or controls. Of note, HLA-G*010401 was found in all aborted cases whereas it was found in 51.9% of succeeded cases (odds ratio = 21.4, P = 0.006, P(c) = 0.03), and the presence of HLA-G*010401 could predict the abortion after PLAT with sensitivity and specificity of 100% and 48.1%, respectively. CONCLUSION: Human leukocyte antigen testing may be useful for predicting effectiveness of PLAT in RSA.

Sex differences of abortuses and neonates in women with allo-immune recurrent abortions.

To investigate the possible association of recurrent spontaneous abortions (RSA) of immune aetiologies with neonatal sex differences, karyotyping of abortuses from allo-immune RSA and epidemiological studies on the sex differences of neonates from sporadic aborters was carried out. Allo-immune disorders, as diagnosed by an increased number of shared HLA class II loci and reduced blocking activity of the woman's serum in mixed lymphocyte reaction, were found almost twice (54.9%) as often as auto-immune disorders (29.9%) among a total of 244 women with RSA. Of 33 abortuses karyotyped from women with RSA, 69.7% showed normal female karyotypes, while only 6.1% had normal male karyotypes, indicating that female fetuses are more prone to abort than males. Epidemiological studies revealed that boys were born at a significantly greater incidence of 58.1% in 221 women with a history of sporadic abortion than 47.6% in 893 women with no history of abortion. Moreover, the proportion of women giving birth to boys only was consistently and significantly higher, regardless of repeated deliveries, in sporadic aborters (36.7%) than in women with no history of abortion (19.6%), showing that more boys were born than girls to women with sporadic abortion. It is concluded that male fetuses are more likely to survive than females in allo-immune RSA due to allo-immune reproductive wastage of chromosomally normal female concept in early human pregnancy, and that allo-immune RSA makes up the highest proportion of unexplained RSA.