Allogeneic bone marrow transplantation with co-stimulatory blockade induces macrochimerism and tolerance without cytoreductive host treatment

Allogeneic bone marrow transplantation (in immunocompetent adults) has always required cytoreductive treatment of recipients with irradiation or cytotoxic drugs to achieve lasting engraftment at levels detectable by non-PCR-based techniques ('macrochimerism' or 'mixed chimerism'). Only syngeneic marrow engraftment at such levels has been achieved in unconditioned hosts. This requirement for potentially toxic myelosuppressive host pre-conditioning has precluded the clinical use of allogeneic bone marrow transplantation for many indications other than malignancies, including tolerance induction. We demonstrate here that treatment of naive mice with a high dose of fully major histocompatibility complex-mismatched allogeneic bone marrow, followed by one injection each of monoclonal antibody against CD154 and cytotoxic T-lymphocyte antigen 4 immunoglobulin, resulted in multi-lineage hematopoietic macrochimerism (of about 15%) that persisted for up to 34 weeks. Long-term chimeras developed donor-specific tolerance (donor skin graft survival of more than 145 days) and demonstrated ongoing intrathymic deletion of donor-reactive T cells. A protocol of high-dose bone marrow transplantation and co-stimulatory blockade can thus achieve allogeneic bone marrow engraftment without cytoreduction or T-cell depletion of the host, and eliminates a principal barrier to the more widespread use of allogeneic bone marrow transplantation. Although efforts have been made to minimize host pre-treatment for allogeneic bone marrow transplantation for tolerance induction, so far none have succeeded in eliminating pre-treatment completely. Our demonstration that this can be achieved provides the rationale for a safe approach for inducing robust transplantation tolerance in large animals and humans.     

How the Electronic Health Record Will Change the Future of Health Care

Genetic testing is expected to play a critical role in patient care in the near future. Advances in genomic research have the potential to impact medicine in very tangible and direct ways, from carrier screening to disease diagnosis and prognosis to targeted treatments and personalized medicine. However, numerous barriers to widespread adoption of genetic testing continue to exist, and health information technology will be a critical means of addressing these challenges. Electronic health records (EHRs) are a digital replacement for the traditional paper-based patient chart designed to improve the quality of patient care. EHRs have become increasingly essential to managing the wealth of existing clinical information that now includes genetic information extracted from the patient genome. The EHR is capable of changing health care in the future by transforming the way physicians use genomic information in the practice of medicine.     

Prevention of suckling improves postpartum reproductive responses to hormone treatments in Pelibuey ewes

To determine the effects of suckling on postpartum (pp) reproductive efficiency in Pelibuey ewes, two experiments were performed. In Experiment 1, 112 ewes were randomly assigned to one of two groups at parturition: Without restriction of suckling (WRS) 24 h day(-1) for 60 days (n=56), and Weaned Ewes (WE), weaned at 40 days pp (n=56). On Day 30 pp, all ewes were given Prostaglandin (PGF2alpha) and one of four treatments (n=14): T1, intravaginal progestagen (FGA; 40 mg) for 12 days from day 30 pp+equine chorionic gonadotropin (eCG; 300 UI) until 2 days before removing FGA; T2, FGA was applied for 12 days; T3, a second application of PGF2alpha was given on day 40 pp+eCG on the same day; T4, a second injection of PGF2alpha was applied on day 40 pp only. In all the analyzed characteristics, the best results were obtained in WE. Within the WE group, the best treatment (P<0.05) was T1 with 85.7% of the ewes in oestrus, 71.4% pregnant and a prolificacy of 1.9. Within the WRS group the best results were observed in T1. In both groups, the lowest results (P<0.05) were obtained in T4. In Experiment 2, 75 ewes were randomly assigned to one of three groups (n=25) immediately after parturition: Group 1, Without restriction of suckling (WRS, as in Experiment 1); Group 2, with restriction of suckling (RS; suckling for 30 min day(-1)); Group 3, Early Weaning (EW: at 7 days pp). All ewes were given PGF2alpha at 30 days pp and the same hormonal treatment, FGA for 12 days+PGF2alpha and eCG 2 days before removing FGA. No differences were observed (P>0.05) between RS and EW for the presentation of oestrus (96% vs. 92%), pregnancy (72% vs. 76%) or prolificacy (1.9 vs. 1.9), although group WRS did not perform (P<0.05) as well as groups RS and EW for any measure of performance. In conclusion, the combination of hormonal treatment (FGA plus eCG) with weaning at 7 or 40 days pp, or restricted suckling, improves postpartum reproductive efficiency in Pelibuey ewes, demonstrating the inhibitory role of suckling on postpartum reproduction in this breed.     

Culture of Tetraselmis tetrathele and its utilization in the hatchery production of different penaeid shrimps in Asia

The prasinophyte Tetraselmis tetrathele is an important live feedorganism for shrimp/prawn hatchery operations because of its highnutritional value and ease of culture. It can replace Brachionus plicatilisand Artemia nauplii as live diet during the protozoeal and mysis feedingstage. This study tried to determine the different optimum cultureconditions for T. tetrathele, and to evaluate its application in thehatchery production of different penaeid species. Tetraselmis tetrathelewas cultured at combinations of six levels of salinities (10–60 gkg^–1), nine levels of pH (3–10.5), and two temperatures(25°C and 30°C) in a three factorialcombination experiment. Effects of varying concentrations of differentorganic and inorganic media including the use of vitamins and trace metalswere also assessed. The dietary value of T. tetrathele was evaluated byfeeding it to different shrimp larvae from protozoea-1 (PZ-1) untilpostlarva-1 (PL-1). There was no significant difference (P > 0.05) inthe growth of T. tetrathele at 25°C and30°C in acidic media; but, there were significantdifferences (P < 0.05) in the range favoring fast growth at25°C. For the inorganic media test, the optimumconcentration for the maximum growth rate of T. tetrathele was at 1 gkg^–1 of Yashima medium without urea (k = 0.57). Thegrowth rate for the other media was highly variable with some approximatingthe optimum. For inorganic nutrients, the optimum concentration for maximumgrowth was at 4 g kg^–1 of unsterilized bio-conversion mediumwith 30 mg l^–1 of Clewat-32 (k = 0.72). The othergrowth rates also indicated significant positive rates but were far from theoptimum. The combination of T. tetrathele and Chaetoceros gracilis or C.calcitrans could be used as the only diet from protozoea-1 (PZ-1) untilpostlarva-1 (PL-1) for Metapenaeus ensis, Metapenaeopsis barbata,Trachypenaeus curvirostris, Penaeus indicus, P. merguiensis, P.latisulcatus, and P. japonicus, with high survival rates. However, thelarvae of P. semisulcatus, P. monodon and P.chinensis can benefit from apure phytoplanton diet until the second mysis stage (M-2) only. Beyond thisstage, the larvae need bigger zooplankton as live feed.     

Cellular Composition of Cerebrospinal Fluid in HIV-1 Infected and Uninfected Subjects

In order to characterize the cellular composition of cerebrospinal fluid (CSF) in a healthy state and in the setting of chronic pleocytosis associated with HIV-1 (HIV) infection, multi-parameter flow cytometry was used to identify and quantitate cellular phenotypes in CSF derived from HIV-uninfected healthy controls and HIV-infected subjects across a spectrum of disease and treatment. CD4+ T cells were the most frequent CSF population and the CD4:CD8 ratio was significantly increased in the CSF compared to blood (p = 0.0232), suggesting preferential trafficking of CD4+ over CD8+ T cells to this compartment. In contrast, in HIV-infection, CD8+ T cells were the major cellular component of the CSF and were markedly increased compared to HIV-uninfected subjects (p<0.001). As with peripheral blood, the CSF CD4:CD8 ratio was reversed in HIV-infected subjects compared to HIV-uninfected subjects. Monocytes, B cells and NK cells were rare in the CSF in both groups, although absolute counts of CSF NK cells and B cells were significantly increased in HIV-infected subjects (p<0.05). Our studies show that T cells are the major cellular component of the CSF in HIV-infected and uninfected subjects. The CSF pleocytosis characteristic of HIV infection involves all lymphocyte subsets we measured, except for CD4+ T cells, but is comprised primarily of CD8+ T cells. The reduced proportion of CD4+ T cells in the CSF may reflect both HIV-related peripheral loss and changes in trafficking patterns in response to HIV infection in the central nervous system.     

Early Developmental Stages of the Green Tiger Prawn, Penaeus semisulcatus de Haan (Crustacea, Decapoda, Penaeidae)

Gravid females of Penaeus semisulcatus were spawned in the laboratory by natural means. The embryos were documented and the larvae were reared from hatching to postlarval stage at 28.2–30.0 °C and 33.5–34.5 g kg⁻¹ salinity for about 10 days (223 h 55 min). Six naupliar stages, three protozoea stages, three mysis stages and the first postlarval stage were described and illustrated. The larvae were fed only with microalgae Tetraselmis tetrathele and Chaetoceros gracilis from first protozoea until the second mysis, with about 90% survival rate; from the third mysis until the first postlarva they were fed with similar microalgae coupled with rotifer Brachionus plicatilis and Artemia nauplii. The embryonic and larval stages of P. semisulcatus are generally similar to those of other closely related species in the family Penaeidae, such as Melicertus canaliculatus, Fenneropenaeus merguiensis, and Marsupenaeus japonicus, except for the size and structure of diagnostic characters, setation of appendages and duration of metamorphoses. The change in the feeding habit during ontogeny was related to morphological transformation of the feeding apparatus of larvae and postlarvae. This paper is the first comprehensive and complete account of the early developmental stages of P. semisulcatus.     

Heterotrimeric Kinesin-2 (KIF3) Mediates Transition Zone and Axoneme Formation of Mouse Photoreceptors

Anterograde intraflagellar transport (IFT) employing kinesin-2 molecular motors has been implicated in trafficking of photoreceptor outer segment proteins. We generated embryonic retina-specific (prefix “emb”) and adult tamoxifen-induced (prefix “tam”) deletions of KIF3a and IFT88 in adult mice to study photoreceptor ciliogenesis and protein trafficking. In ^embKif3a^−/− and in ^embIft88^−/− mice, basal bodies failed to extend transition zones (connecting cilia) with outer segments, and visual pigments mistrafficked. In contrast, ^tamKif3a^−/− and ^tamIft88^−/− photoreceptor axonemes disintegrated slowly post-induction, starting distally, but rhodopsin and cone pigments trafficked normally for more than 2 weeks, a time interval during which the outer segment is completely renewed. The results demonstrate that visual pigments transport to the retinal outer segment despite removal of KIF3 and IFT88, and KIF3-mediated anterograde IFT is responsible for photoreceptor transition zone and axoneme formation.