The effects of Bombyx mori silk strain and extraction time on the molecular and biological characteristics of sericin

Sericin was extracted from three strains of Thai Bombyx mori silk cocoons (white shell Chul1/1, greenish shell Chul3/2, and yellow shell Chul4/2) by a high-pressure and high-temperature technique. The characteristics of sericin extracted from different fractions (15, 45, and 60 min extraction process) were compared. No differences in amino acid composition were observed among the three fractions. For all silk strains, sericin extracted from a 15-min process presented the highest molecular weight. The biological potential of the different sericin samples as a bioadditive for 3T3 fibroblast cells was assessed. When comparing sericin extracted from three silk strains, sericin fractions extracted from Chul4/2 improved cell proliferation, while sericin from Chul 1/1 activated Type I collagen production to the highest extent. This study allows the natural variability of sericin obtained from different sources and extraction conditions to be addressed and provides clues for the selection of sericin sources.     

Two new flavonoids from Derris laxiflora Benth

Two new compounds, derriscoumaronochromone (1) and cis-3,4′-dihydroxy- 5,7-dimethoxyflavan (2), as well as trans-4'-O-methylcatechin (3) were isolated from Derris laxiflora, The structures of these compounds were determined by analysis of their spectroscopic data.     

Determination of field output correction factors of radiophotoluminescence glass dosimeter and CC01 ionization chamber and validation against IAEA-AAPM TRS-483 code of practice

PurposeTo determine the field output correction factors of the radiophotoluminescence glass dosimeter (RPLGD) in parallel and perpendicular orientations with reference to CC01, the ionization chamber.MethodsThe dose to a small water volume and the sensitive volume of the RPLGD and the IBA-CC01 were determined for 6-MV, 100-cm SAD, 10-cm depth using egs_chamber user-code. The RPLGD in perpendicular and parallel orientations to the beam axis were studied. The field output correction factors of each detector for 0.5 × 0.5 to 10 × 10 cm² field sizes were determined. These field output correction factors were validated by comparing field output factors against data determined from IAEA-AAPM TRS-483 code of practice.ResultsThe field output correction factors of all detectors were within 5% for field sizes down to 0.8 × 0.8 cm². For 0.5 × 0.5 cm², the field output correction factors of CC01, RPLGD in perpendicular and parallel orientations differed from unity by 14%, 19%, and 5%, respectively. The percentage difference between field output factors determined using RPLGD and CC01 data, corrected using the field output correction factors determined in this work and measurements with CC01 data corrected using TRS-483, was less than 3% for all field sizes, except for the smallest field size of RPLGD in perpendicular orientation and the CC01.ConclusionsThe field output correction factors of RPLGD and CC01 are reported. The validation proves that RPLGD in parallel orientation combined with the field output correction factors is the most suitable for determining the field output factors for the smallest field used in this study.     

The inhibitory activity of aldose reductase in vitro by constituents of Garcinia mangostana Linn

We investigated aldose reductase inhibition of Garcinia mangostana Linn. from Indonesia. Dichloromethane extract of the root bark of this tree was found to demonstrate an IC₅₀ value of 11.98 µg/ml for human aldose reductase in vitro. From the dichloromethane fraction, prenylated xanthones were isolated as potent human aldose reductase inhibitors. We discovered 3-isomangostin to be most potent against aldose reductase, with an IC₅₀ of 3.48 µM.     

Genetic diversity assessment and estimation of phylogenetic relationships among 26 Fagaceae species using ISSRs

The family Fagaceae includes several species and presents huge genetic variability. In the last two decades, several genetic studies about phylogenetics and genetic diversity of Fagaceae have emerged. ISSR markers were used to evaluate the genetic diversity of 26 species of Fagaceae belonging to the genera Castanea, Fagus and Quercus. Among several primers tested, 17 were selected for the evaluation of diversity and estimation of genetic relationships. A total of 371 ISSR markers were produced and each primer revealed high polymorphism. Specific ISSR markers for the Quercus infrageneric groups were amplified. ISSRs proved to be a reliable tool for the discrimination of the analyzed species per genus, infrageneric group and/or ecological origin.     

Biology of nitrogen-fixing Rhizobacteria

In non-legumes associative nitrogen-fixing system, several genera of rhizobacteria have been reported. The object of this paper is to summarize the current understanding of how rhizobacteria adhere to the root surface of non-legumes especially rice and other cereal crops. Evidence for involvement of rice lectin in adhesion will be reviewed. An emphasis will be placed on theKlebsiella R15 ammonium assimilation system in free-living state and in associative state with rice seedlings. Nitrogenase and glutamine synthetase (GS) activities of associativeKlebsiella increased significantly in the rhizosphere of rice comparing to the free-living state. In rice, the soluble form of GS specific activity appear to be slightly lower than in rice root in the absence of bacteria. These results suggest that nitrogen-fixing activity has been enhanced during association. The dinitrogen fixed should be changed to amino acids via GS-GOGAT pathway in bacteria. Transfer of fixed nitrogen and assimilation in the rice plant is the problem that needs to be solved in order to improve the efficiency of associative nitrogen fixation.     

A remarkable new Bulbophyllum, sect. Sestochilus (Orchidaceae) from Thailand

Ongoing studies show that what hitherto in Thailand and neighboring countries have been gathered under one variable species, Bulbophyllum lobbii , must be separated in a number of distinct, well defined species. One such. B. smitinandii , is treated here.     

Manufacturing and validation of Good Manufacturing Practice–compliant regulatory dendritic cells for infusion into organ transplant recipients

Background aimsRegulatory (or “tolerogenic”) dendritic cells (DCregs) are a highly promising, innovative cell therapy for the induction or restoration of antigen-specific tolerance in immune-mediated inflammatory disorders. These conditions include organ allograft rejection, graft-versus-host disease following bone marrow transplantation and various autoimmune disorders. DCregs generated for adoptive transfer have potential to reduce patients’ dependence on non-specific immunosuppressive drugs that can induce serious side effects and enhance the risk of infection and certain types of cancer. Here, our aim was to provide a detailed account of our experience manufacturing and validating comparatively large numbers of Good Manufacturing Practice–grade DCregs for systemic (intravenous) infusion into 28 organ (liver) transplant recipients and to discuss factors that influence the satisfaction of release criteria and attainment of target cell numbers.ResultsDCregs were generated in granulocyte-macrophage colony stimulating factor and interleukin (IL)-4 from elutriated monocyte fractions isolated from non-mobilized leukapheresis products of consenting healthy adult prospective liver transplant donors. Vitamin D3 was added on day 0 and 4 and IL-10 on day 4 during the 7-day culture period. Release and post-release criteria included cell viability, purity, phenotype, sterility and functional assessment. The overall conversion rate of monocytes to DCregs was 28 ± 8.2%, with 94 ± 5.1% product viability. The mean cell surface T-cell co-inhibitory to co-stimulatory molecule (programmed death ligand-1:CD86) mean fluorescence intensity ratio was 3.9 ± 2.2, and the mean ratio of anti-inflammatory:pro-inflammatory cytokine product (IL-10:IL-12p70) secreted upon CD40 ligation was 60 ± 63 (median = 40). The mean total number of DCregs generated from a single leukapheresis product (n = 25 donors) and from two leukapheresis products (n = 3 donors) was 489 ± 223 × 10⁶ (n = 28). The mean total number of DCregs infused was 5.9 ± 2.8 × 10⁶ per kg body weight. DCreg numbers within a target cell range of 2.5–10 × 10⁶/kg were achieved for 25 of 27 (92.6%) of products generated.ConclusionsHigh-purity DCregs meeting a range of quality criteria were readily generated from circulating blood monocytes under Good Manufacturing Practice conditions to meet target cell numbers for infusion into prospective organ transplant recipients.     

Day 4 collection of granulocyte colony-stimulating factor-mobilized HLA-matched sibling donor peripheral blood allografts demonstrates no long-term increase in chronic graft-versus-host disease or relapse rates

Background aimsIn a previous pilot study of HLA-matched sibling donor hematopoietic cell transplantation (HCT), the authors determined the feasibility of day 4 versus day 5 granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood stem cell (PBSC) collection compared with a historical cohort. Given identified differences in the PBSC product (day 4 cohort with significantly lower infused total nucleated, mononuclear and CD3 cells compared with other collection cohorts), the authors performed a follow-up study to determine long-term post-HCT outcomes, including detailed characterization of chronic graft-versus-host disease (GVHD).MethodsThis was a prospective observational study, and the authors collected data on chronic GVHD, staging, sites of involvement and treatments. Performance status, incidence of relapse, overall survival and duration of immunosuppressive therapy (IST) were also evaluated. Data were examined retrospectively. To account for differences in length of follow-up among cohorts, the authors also determined performance status and chronic GVHD staging, sites and treatment at 2 years post-HCT.ResultsAt 2 years post-HCT, the overall survival rate was 71.7% in the day 4 cohort compared with 61.5%, 52% and 56% in the day 5, 2-day and historical cohorts, respectively (P = 0.283). The cumulative incidence of chronic GVHD was 65.2% in the day 4 cohort versus 46.4% in the day 5 cohort, 51.1% in the 2-day cohort and 65% in the historical cohort (P = 0.26). There was no significant difference in the maximum overall stage of chronic GVHD (P = 0.513), median number of sites involved (P = 0.401) or cumulative incidence of discontinuation of IST (P = 0.32). Death from chronic GVHD was less common in the day 4 and day 5 cohorts compared with the 2-day and historical cohorts, though this did not reach statistical significance.ConclusionsThe authors’ preliminary results demonstrated that collection of allogeneic matched sibling donor PBSCs on day 4 of G-CSF was feasible, reduced donor exposure to growth factor and was associated with an initial cost savings. Importantly, the authors now demonstrate that transplantation of day 4 mobilized PBSCs is not associated with any adverse outcomes post-HCT, including late effects such as chronic GVHD. Further investigation of donor G-CSF collection algorithms is merited in other HCT settings, including unrelated and mismatched related donors.