Total synthesis of exigurin: the Ugi reaction in a hypothetical biosynthesis of natural products.

The first total synthesis of a marine natural product, exigurin, has been accomplished in 13 steps starting from (+)-menthone. The key intermediate (-)-10-epi-axisonitrile-3 was prepared by stereoselective intramolecular cyclopropanation followed by a cyclopropane ring opening reaction by the azide anion. The bioinspired Ugi reaction of (-)-10-epi-axisonitrile-3, formaldehyde, sarcosine and methanol successfully constructed the target exigurin in which its terpene and amino acid units were linked through an amide bond.

Asymmetric organocatalytic desymmetrization of 4,4-disubstituted cyclohexadienones at high pressure: a new powerful strategy for the synthesis of highly congested chiral cyclohexenones.

A highly diastereoselective and enantioselective method for the asymmetric desymmetrization of 4,4-disubstituted cyclohexadienones using the Michael addition reaction of malonates under catalysis with the primary amine-thiourea conjugate catalyst and PPY at high pressure was developed.

Protecting group free synthesis of urea-linked glycoconjugates: efficient synthesis of ß-urea glycosides in aqueous solution.

A method for the protecting group free synthesis of ß-urea-linked glycoconjugates has been developed. The one step process, involving reactions between urea and D-glucose, N-acetyl-D-glucosamine or D-xylose in acidic aqueous solution, furnishes the corresponding ß-urea glycosides in modest yields. This simple and efficient procedure is applicable to the synthesis of ß-urea tethered amino acid-carbohydrate conjugates.

Home-based oral immunotherapy (OIT) with an intermittent loading protocol in children unlikely to outgrow egg allergy.

BACKGROUND: Home based oral immunotherapy (OIT) for food allergy has often been used for young children in Japan, the majority of whom are believed to outgrow the allergy by the school age, therefore the true efficacy of the therapy has been controversial. The aim of this study was to evaluate the efficacy and safety of a newly developed slow- type home-based oral immunotherapy (OIT) regimen in children with hen's egg (HE) allergy, who had low likelihood of outgrowing the allergy, with treatment involving only elimination diet. METHOD: We retrospectively reviewed the medical records of 43 children with egg allergy (30 males; median age 6) who fulfilled Burks et al.'s criteria of being unlikely to outgrow the allergy. Thirty children who agreed to start OIT were assigned to the treatment group, and 13 who did not want to participate immediately were assigned to the untreated group; the patients underwent an elimination diet for 1 year, during which they were monitored. The OIT regimen involved the intake of the maximum tolerated dose 2 to 3 times a week at home, with initial dose introduction followed by dose build-ups with medical supervision. We statistically evaluated the rate of children who changed their threshold up to 32 g of egg - defined as, oral tolerance induction- in both the groups for 1 year and in the OIT group for 2 years, as well as the rate of children who fulfilled Savage et al.'s criteria of clinical tolerance after reaching the abovementioned remission stage. RESULTS: The rate of children who achieved oral tolerance induction to 32 g of egg after 1 year in the OIT group (9/30) was significantly higher than that in the untreated group (0/13). The total rate within the OIT group was significantly increased from 9/30 at 1 year to 17/30 at two years without any severe adverse reaction; of the above 17 children, we followed 14 children, and noted that 11 of these were able to obtain clinical tolerance. CONCLUSION: The home-based OIT with an intermittent loading protocol was very safe and effective in children with a low likelihood of outgrowing egg allergy.

Predictive value of IgE/IgG4 antibody ratio in children with egg allergy.

BACKGROUND: The aim of this study was to investigate the role of specific IgG4 antibodies to hen's egg white and determine their utility as a marker for the outcome of oral challenge test in children sensitized to hen's egg METHODS: The hen's egg oral food challenge test was performed in 105 sensitized children without atopic dermatitis, and the titers of egg white-specific immunoglobulin G4 (IgG4) and immunoglobulin E (IgE) antibodies were measured. To set the cut-off values of IgG4, IgE, and the IgE/IgG4 ratio for predicting positive results in oral challenges, receiver operating characteristic curves were plotted and the area under the curves (AUC) were calculated. RESULTS: Sixty-four of 105 oral challenges with whole eggs were assessed as positive. The AUC for IgE, IgG4, and IgE/IgG4 for the prediction of positive results were 0.609, 0.724, and 0.847, respectively. Thus, the IgE/IgG4 ratio generated significantly higher specificity, sensitivity, positive predictive value (%), and negative predictive value (%) than the individual IgE and IgG4. The negative predictive value of the IgE/IgG4 ratio was 90% at a value of 1. CONCLUSIONS: We have demonstrated that the egg white-specific serum IgE/IgG4 ratio is important for predicting reactivity to egg during food challenges.

Aminohydroxyacetone synthons: versatile intermediates for the organocatalytic asymmetric aldol reaction.

A practical method for the synthesis of 1,3-aminohydroxyacetone synthons was developed, and their utility in the organocatalytic asymmetric aldol reaction was demonstrated in a short synthesis of aza-sugars.

Synthesis of manzacidin A and C: efficient construction of quaternary carbon stereocenters bearing nitrogen substituents.

An efficient synthetic method for stereoselective construction of asymmetric quaternary carbon stereocenters, bearing nitrogen in the form of Boc-protected allyl amines, has been developed. This methodology is employed in the synthesis of marine alkaloids, manzacidin A and C.

Asymmetric synthesis of (+)-geranyllinaloisocyanide: assignment of absolute stereochemistry.

The first nonracemic synthesis of (+)-geranyllinaloisocyanide, starting with (-)-lactic acid methyl ester, has been accomplished by exploiting a [3.3] sigmatropic rearrangement of allyl cyanate. The synthesis enables assignment of the S configuration of the C(3) isocyano substituted, quaternary stereogenic center in natural geranyllinaloisocyanide.

A new powerful strategy for the organocatalytic asymmetric construction of a quaternary carbon stereogenic center.

A new method for chiral diamine-catalyzed Robinson-type annulation was developed to construct cyclohexenone derivatives bearing a quaternary carbon stereogenic center at the 4-position in high enantiomeric excess. This method was successfully applied to the short synthesis of (+)-sporochnol A.

Synthesis of glycocinnasperimicin D.

The first total synthesis of amino sugar antibiotic glycocinnasperimicin D (1) has been achieved by a convergent, three-component coupling strategy. The key steps involve the Heck-Mizoroki reaction by using the iodophenyl glycoside 50 and acryl amide 32 to furnish the right core structure of 1, and the construction of the urea glycoside employing the reaction of glycosyl isocyanate 8 with amino sugar 9. Glycosyl isocyanate 8 was prepared by the oxidation of isonitrile 10, which displayed excellent reactivity in the coupling event. Synthetic roadblocks, encountered during this synthetic effort, have led to the development of the alpha-selective, Lewis acid catalyzed phenyl glycosylation process with 2-amino-hexopyranose and a procedure for acetonide deprotection without affecting the silyl ethers.

[Quality and performance of pap smears in the cervical cancer screening program in a city of southern Brazil]. / Qualidade e desempenho das colpocitologias na prevenção de câncer de colo uterino.

OBJECTIVE: To estimate prevalence and analyze quality and performance of colpocytology carried out under the Cervical Cancer Screening Program in Maringá, Paraná, Brazil. METHODS: A retrospective study of the SIS-Colo database of the Brazilian Ministry of Health. Variables such as age, colpocytology result and origin were analyzed. Ages were divided into brackets; origin was distributed in five Regional Health Divisions of the city of Maringá; colpocytology was categorized according to the Bethesda System. Colpocytology, coverage was calculated by dividing the number of exams in the population between 25 and 59 years of age by the number of women in that same age group. RESULTS: The 17,664 colpocytology exams collected in 2005 by the public health system were distributed among women between 12 and 82 years of age, with 12,961 (73.4%) examinations in women between the ages of 25 and 59, considered at high risk for cancer. A total of 17,458 (98.84%) cytological examinations were negative for malignancy, and atypical results (ASCUS / AGUS, LSIL, HSIL and invasive cancer) totaled 206 (1.16%). The study found a prevalence of 0.85% (151) for ASCUS / AGUS, 1.14% (203/17.664) for atypical cells, and a 2.75 ASCUS / atypical cells ratio (151/55). CONCLUSION: The lower than expected prevalence of colpocytological and ASCUS alterations, the high ASCUS / atypical cells ratio and the insufficient population coverage by colpocytology jeopardized the performance of the cervical cancer prevention program. The low-income population requires special attention, and those more privileged should be advised about the frequency of examination and the age bracket with higher risks.

Qualidade e desempenho das colpocitologias na prevenção de câncer de colo uterino / Quality and performance of pap smears in the cervical cancer screening program in a city of southern Brazil

OBJETIVO: Estimar a prevalência e analisar a qualidade e o desempenho das colpocitologias realizadas no Programa de Prevenção de Câncer de Colo Uterino no município de Maringá-PR. MÉTODOS: Estudo retrospectivo dos dados do SIS-Colo do Ministério da Saúde. Foram analisadas as variáveis idade, resultado de colpocitologia e procedência. As idades foram categorizadas em faixas etárias, a procedência distribuída em cinco regionais de saúde do município de Maringá; a colpocitologia foi categorizada conforme o Sistema Bethesda. Calculou-se a cobertura de colpocitologia dividindo-se o número de exames na população de 25 a 59 anos pelo número de mulheres nesta mesma faixa etária. RESULTADOS: As 17.664 colpocitologias coletadas, no ano de 2005, pela rede pública se distribuíram entre as mulheres com idade mínima de 12 e máxima de 82 anos, sendo 12.961 (73,4 por cento) exames na faixa entre 25 e 59 anos, considerada de maior risco para o câncer. Foram negativos para neoplasia 17.458 (98,84 por cento) exames e as alterações celulares (ASCUS/AGUS, LIEBG, LIEAG e câncer invasor) totalizaram 206 (1,16 por cento). Encontrou-se prevalência de ASCUS de 0,85 por cento (151), de atipias celulares 1,14 por cento (203/17.664) e a relação ASCUS/atipias celulares 2,75 por cento (151/55). CONCLUSÃO: A prevalência das alterações colpocitológicas e da ASCUS abaixo dos índices esperados, a relação ASCUS/atipias celulares em níveis elevados, a cobertura populacional insuficiente de colpocitologia compromete o desempenho da prevenção de câncer de colo uterino. A população de baixa condição sócio-econômica requer uma atenção especial e a população mais privilegiada deve ser orientada quanto a periodicidade do exame e faixa etária de maior risco.

OBJECTIVE: To estimate prevalence and analyze quality and performance of colpocytology carried out under the Cervical Cancer Screening Program in Maringá, Paraná, Brazil. METHODS: A retrospective study of the SIS-Colo database of the Brazilian Ministry of Health. Variables such as age, colpocytology result and origin were analyzed. Ages were divided into brackets; origin was distributed in five Regional Health Divisions of the city of Maringá; colpocytology was categorized according to the Bethesda System. Colpocytology, coverage was calculated by dividing the number of exams in the population between 25 and 59 years of age by the number of women in that same age group. RESULTS: The 17,664 colpocytology exams collected in 2005 by the public health system were distributed among women between 12 and 82 years of age, with 12,961 (73.4 percent) examinations in women between the ages of 25 and 59, considered at high risk for cancer. A total of 17,458 (98.84 percent) cytological examinations were negative for malignancy, and atypical results (ASCUS / AGUS, LSIL, HSIL and invasive cancer) totaled 206 (1.16 percent). The study found a prevalence of 0.85 percent (151) for ASCUS / AGUS, 1.14 percent (203/17.664) for atypical cells, and a 2.75 ASCUS / atypical cells ratio (151/55). CONCLUSION: The lower than expected prevalence of colpocytological and ASCUS alterations, the high ASCUS / atypical cells ratio and the insufficient population coverage by colpocytology jeopardized the performance of the cervical cancer prevention program. The low-income population requires special attention, and those more privileged should be advised about the frequency of examination and the age bracket with higher risks.

Administration of granulocyte colony-stimulating factor to recipients followed by intra-bone marrow-bone marrow transplantation accelerates acceptance of allogeneic bone marrow cells in mice.

We have recently established a novel method for bone marrow transplantation: intra-bone marrow-bone marrow transplantation (IBM-BMT), by which the rapid recovery of donor-derived hematopoiesis can be expected even when reduced radiation doses are used. In this paper, we examine, using mice, whether the combination of pretreatment of recipients with granulocyte-colony-stimulating factor (G-CSF) and IBM-BMT can induce a more rapid recovery of donor-derived hematopoiesis than IBM-BMT alone. We first pretreated recipients with recombinant human (rh) G-CSF (250 microg/kg/day) for 5 consecutive days (days -6 to -2). On day -1, the recipients were irradiated, and IBM-BMT was carried out on day 0. On day 12, we performed colony-forming units of spleen (CFU-S) assays. The combination of G-CSF pretreatment and IBM-BMT augmented the CFU-S counts, the weight of spleens, and the numbers of donor-derived hematopoietic cells. We next analyzed the mechanisms underlying these effects of G-CSF and found that (i) G-CSF induces Th2 polarization, which can prevent graft rejection, and (ii) G-CSF augments natural suppressor activity, which suppresses graft rejection. The combination of G-CSF pretreatment and IBM-BMT can produce the rapid recovery of donor-derived hematopoiesis and suppress graft rejection. This method would lighten the burden on patients in allogeneic BMT.

Synthesis of (-)-agelastatin A by [3.3] sigmatropic rearrangement of allyl cyanate.

Total synthesis of (-)-agelastatin A has been achieved starting from l-arabitol. The highlights in our synthesis include the preparation of vicinal diamine moiety by [3.3] sigmatropic rearrangement of allyl cyanate and construction of central ring-C with ring-closing metathesis.

Immunotherapy for malignant tumors using combination of allogeneic intra-bone marrow-bone marrow transplantation, donor lymphocyte infusion and dendritic cells.

We have previously shown that the combination of allogeneic intra-bone marrow-bone marrow transplantation (IBM-BMT) and donor lymphocyte infusion (DLI) using CD4+ cell-depleted spleen cells is effective in suppressing tumor growth, but that this does not induce graft-versus-host disease (GVHD) in mice. In this report, we show that formalin-fixed tumor cell-pulsed dendritic cells (FFTCP DCs) have an additive effect with IBM-BMT plus DLI on the suppression of tumor growth, but that the DCs do not augment GVHD. BALB/c mice, which had been subcutaneously inoculated with Meth A (BALB/c-derived fibrosarcoma), were irradiated at a low dose (5 Gy) and were transplanted with bone marrow cells (BMCs) from C57BL/6 (B6) mice into the bone marrow cavity (IBM-BMT). Simultaneously, the mice were intravenously injected with spleen cells from B6 mice, and subcutaneously injected with FFTCP DCs derived from the bone marrow (BM) of B6 mice. At the point of the induction of DCs from BMCs, formalin-fixed Meth A cells were added into the culture. The mice treated with the combination of FFTCP DCs, IBM-BMT and DLI using CD4+ cell-depleted spleen cells showed smaller tumor sizes and longer survival than the mice treated with IBM-BMT plus FFTCP DCs or IBM-BMT plus DLI using CD4+ cell-depleted spleen cells. These results suggest that the combination of FFTCP DCs, IBM-BMT plus DLI using CD4+ cell-depleted spleen cells has potent anti-tumor effects without showing GVHD.

Allogeneic intrabone marrow-bone marrow transplantation plus donor lymphocyte infusion suppresses growth of colon cancer cells implanted in skin and liver of rats.

We have recently found that allogeneic intrabone marrow-bone marrow transplantation (IBM-BMT) + donor lymphocyte infusion (DLI) using CD4(+) cell-depleted spleen cells (CD4(-) cells) can prevent graft-versus-host disease (GvHD) but suppress tumor growth (Meth A: fibrosarcoma) in mice. In the present study, we show that allogeneic IBM-BMT + DLI using CD4(-) cells also has suppressive effects on the growth of colon cancer cells implanted not only in the skin but also in the liver of rats. First, we examined the effects of allogeneic IBM-BMT + DLI on the subcutaneously inoculated ACL-15 (rat colon cancer cell line). Lethally irradiated Fischer rats (F344 rats) were transplanted with T-cell-depleted bone marrow cells (BMCs) from Brown Norway (BN) rats. Simultaneously, DLI was performed using whole spleen cells (whole cells), CD4(+) cell-depleted spleen cells (CD4(-) cells) or CD8(+) cell-depleted spleen cells (CD8(-) cells) of BN rats. Although allogeneic IBM-BMT + DLI suppressed tumor growth, a considerable number of rats treated with allogeneic IBM-BMT + DLI using whole cells or CD8(-) cells died due to GvHD. In contrast, allogeneic IBM-BMT + DLI using CD4(-) cells also suppressed tumor growth, but there was no GvHD. Based on these findings, we next examined the effects of allogeneic IBM-BMT + DLI using CD4(-) cells on the cancer cells implanted in the liver. Allogeneic IBM-BMT + DLI using CD4(-) cells via the portal vein significantly prolonged the survival. These results suggest that allogeneic IBM-BMT + DLI using CD4(-) cells could become a new strategy for the treatment of solid tumors.

An innovative approach to bone marrow collection and transplantation in a patient with beta-thalassemia major: marrow collection using a perfusion method followed by intra-bone marrow injection of collected bone marrow cells.

Using small animals (mice and rats) and monkeys, we have found that the combination of bone marrow collection using the perfusion method (PM) and intra-bone marrow-bone marrow transplantation (IBM-BMT) of the collected cells is safe and effective in treating various intractable diseases. Based on these findings, we attempted to apply this method to humans. We report here the first case of a patient (6 years old) with beta-thalassemia major who underwent allogeneic BMT using this new PM + IBM-BMT method. The white blood cell counts of the patient gradually increased to more than 1500/microL by day 47 and continued to increase, reaching the highest level (8600/microL) on day +55. Fluorescence in situ hybridization data on day +33 showed that 98% of the peripheral blood cells were from the donor. Notably, there were no symptoms of graft-versus-host disease (GvHD). However, on day +56, the patient regrettably died of asphyxia resulting from sticky sputum. There was no evidence of infection (in the lung or liver) or GvHD (in the skin) by necropsy. We hope that this case report will help make our new strategies more readily available for the treatment of patients with various intractable diseases.