Dendritic cell phenotype in severe asthma reflects clinical responsiveness to glucocorticoids.

BACKGROUND: Subsets of patients with severe asthma remain symptomatic despite prolonged, high-dose glucocorticoid therapy. We hypothesized that the clinical glucocorticoid sensitivity of these asthmatics is reflected in differences in peripheral blood dendritic cell subsets. OBJECTIVE: To compare peripheral blood leucocyte populations using flow cytometry at baseline and after 2 weeks of systemic glucocorticoid (steroid) treatment to identify immunological differences between steroid-sensitive (SS) and steroid-resistant (SR) asthmatics. METHODS: Adult severe asthmatics (SS n = 12; SR n = 23) were assessed for their response to 2 weeks of therapy with oral prednisolone. Peripheral blood was obtained before and after therapy and stained for lymphocyte (CD3, CD19, CD4, CD8 and Foxp3) and dendritic cell markers (Lineage negative [CD3, CD14, CD16, CD19, CD20, CD56], HLA-DR+, CD304, CD11c, ILT3 and CD86). RESULTS: A higher median frequency of myeloid DCs (mDCs) but not plasmacytoid DCs (pDCs) was observed in the blood of SR as compared to SS asthmatics (P = .03). Glucocorticoid therapy significantly increased median B cell, but not T cell numbers in both cohorts, with a trend for increased numbers of Foxp3+ Tregs in SS (P = .07), but not SR subjects. Oral prednisolone therapy significantly reduced the median numbers and frequencies of total DCs and pDCs in both SS and SR asthmatics. Interestingly, the expression of HLA-DR and ILT3 was also reduced on pDCs in all patients. In contrast, therapy increased the median frequency of mDCs in SS, but reduced it in SR asthmatics. CONCLUSIONS: Myeloid DC frequency is elevated in SR compared with SS asthmatics, and mDC shows a differential response to oral prednisolone therapy.

Half adder capabilities of a coupled quantum dot device.

In this paper we demonstrate two realizations of a half adder based on a voltage-rectifying mechanism involving two Coulomb-coupled quantum dots. First, we examine the ranges of operation of the half adder's individual elements, the AND and XOR gates, for a single rectifying device. It allows a switching between the two gates by a control voltage and thus enables a clocked half adder operation. The logic gates are shown to be reliably operative in a broad noise amplitude range with negligible error probabilities. Subsequently, we study the implementation of the half adder in a combined double-device consisting of two individually tunable rectifiers. We show that this double device allows a simultaneous operation of both relevant gates at once. The presented devices draw their power solely from electronic fluctuations and are therefore an advancement in the field of energy efficient and autonomous electronics.

Voltage fluctuation to current converter with Coulomb-coupled quantum dots.

We study the rectification of voltage fluctuations in a system consisting of two Coulomb-coupled quantum dots. The first quantum dot is connected to a reservoir where voltage fluctuations are supplied and the second one is attached to two separate leads via asymmetric and energy-dependent transport barriers. We observe a rectified output current through the second quantum dot depending quadratically on the noise amplitude supplied to the other Coulomb-coupled quantum dot. The current magnitude and direction can be switched by external gates, and maximum output currents are found in the nA region. The rectification delivers output powers in the pW region. Future devices derived from our sample may be applied for energy harvesting on the nanoscale beneficial for autonomous and energy-efficient electronic applications.

A 1-year randomized, double-blind, placebo-controlled study of intravenous ibandronate on bone loss following renal transplantation.

The clinical profile of ibandronate as add-on to calcitriol and calcium was studied in this double-blind, placebo-controlled trial of 129 renal transplant recipients with early stable renal function (≤ 28 days posttransplantation, GFR ≥ 30 mL/min). Patients were randomized to receive i.v. ibandronate 3 mg or i.v. placebo every 3 months for 12 months on top of oral calcitriol 0.25 mcg/day and calcium 500 mg b.i.d. At baseline, 10 weeks and 12 months bone mineral density (BMD) and biochemical markers of bone turnover were measured. The primary endpoint, relative change in BMD for the lumbar spine from baseline to 12 months was not different, +1.5% for ibandronate versus +0.5% for placebo (p = 0.28). Ibandronate demonstrated a significant improvement of BMD in total femur, +1.3% versus -0.5% (p = 0.01) and in the ultradistal radius, +0.6% versus -1.9% (p = 0.039). Bone formation markers were reduced by ibandronate, whereas the bone resorption marker, NTX, was reduced in both groups. Calcium and calcitriol supplementation alone showed an excellent efficacy and safety profile, virtually maintaining BMD without any loss over 12 months after renal transplantation, whereas adding ibandronate significantly improved BMD in total femur and ultradistal radius, and also suppressed biomarkers of bone turnover. Ibandronate was also well tolerated.

The Barts Health NHS Trust COVID-19 cohort: characteristics, outcomes and risk scoring of patients in East London.

BACKGROUND: Barts Health National Health Service Trust (BHNHST) serves a diverse population of 2.5 million people in London, UK. We undertook a health services assessment of factors used to evaluate the risk of severe acute respiratory coronavirus 2 (SARS-CoV-2) infection.METHODS: Patients with confirmed polymerase chain reaction (PCR) test results admitted between 1 March and 1 August 2020 were included, alongwith clinician-diagnosed suspected cases. Prognostic factors from the 4C Mortality score and 4C Deterioration scores were extracted from electronic health records and logistic regression was used to quantify the strength of association with 28-day mortality and clinical deterioration using national death registry linkage.RESULTS: Of 2783 patients, 1621 had a confirmed diagnosis, of whom 61% were male and 54% were from Black and Minority Ethnic groups; 26% died within 28 days of admission. Mortality was strongly associated with older age. The 4C mortality score had good stratification of risk with a calibration slope of 1.14 (95% CI 1.01-1.27). It may have under-estimated mortality risk in those with a high respiratory rate or requiring oxygen.CONCLUSION: Patients in this diverse patient cohort had similar mortality associated with prognostic factors to the 4C score derivation sample, but survival might be poorer in those with respiratory failure.

Aspects of present and future data presentation in Scandiatransplant.

Scandiatransplant is the Nordic organ exchange organization having existed for almost 40 years. With close collaboration between transplant centers in the Nordic countries, it has been valuable to ensure the optimal usage of available organs. The heart is the most often exchanged organ within the collaboration. It has been decided to create a priority for hyperimmunized kidney patients for compulsory exchange of organs from deceased donors. The age of the deceased organ donors has changed from younger to older donors. The evaluation of deceased kidney transplantations and deceased liver transplantations from 1995 to 2007 is shown for 4 countries. Iceland by itself is performing living donor kidney transplantations with great intensity. Scandiatransplant will make efforts to present more data than just transplantation to yield a more complete picture of organ transplantation.

Early, abrupt conversion of de novo renal transplant patients from cyclosporine to everolimus: results of a pilot study.

In a single-center study, 20 kidney transplant patients without prior rejection were abruptly converted from cyclosporine to everolimus at seven wk post-transplant. All patients received basiliximab induction with maintenance enteric-coated mycophenolate sodium and corticosteroids. Biopsy-proven acute rejection had occurred in three of 20 patients (15.0%) by the end of week seven post-conversion. All episodes were mild and reversible, with subsequent recovery of renal function. Calculated glomerular filtration rate (GFR) improved significantly (51 +/- 11 mL/min at time of conversion, 58 +/- 12 mL/min at week seven post-conversion, 57 +/- 17 mL/min at month six post-conversion; p = 0.001). No patient developed proteinuria in the nephrotic range. Twenty-two adverse events were reported in 10 patients, three of which had a suspected relationship to everolimus. Mean leukocyte and platelet count decreased significantly, and triglyceride level increased. This study suggests that kidney transplant patients without prior rejection can be converted abruptly from cyclosporine to everolimus at seven wk post-transplant, resulting in significantly improved renal function with no apparent increase, in risk of rejection and good tolerability.

Expression of activin pathway genes in granulosa cells of dominant and subordinate bovine follicles.

We examined the association between the expression profiles of genes of the activin signalling pathway and ovarian follicular dominance in cattle. In monovular species such as cattle, one ovarian follicle of a cohort is selected to become dominant, whereas all others (i.e. the subordinate follicles) eventually succumb to apoptosis. We showed that Inhibin-betaA, coding for the betaA chain found in the A isoforms of activin, Inh-alpha coding for the inhibin-specific alpha chain, and the activin antagonist follistatin were expressed at higher levels in dominant follicle granulosa cells from Day 3.5 (ovulation=Day 0). Before selection, Inh-betaA but not Inh-alpha was significantly correlated with potential dominant follicles, as defined by high aromatase expression and follicular fluid estrogen concentrations. Follistatin expression marked the largest follicles at Day 1.5, but displayed large variation in levels among cows. The third inhibin gene, Inh-betaB, could only be detected at very low levels from Day 7 and thus was unlikely to play a prominent role in activin/inhibin signalling in cattle during these stages. There was a decrease in activin tone (P=0.07) specifically in the aromatase-high/dominant follicles, as measured by the ratio of Inh-betaA to Inh-alpha plus follistatin transcripts between Days 1.5 and 7. Messenger RNA for both activin type II receptors and the nuclear effector Smad2 were detected in granulosa cells, consistent with an autocrine role for activin signalling. Additionally, expression of the putative activin target genes Smad2 and FSHreceptor were, respectively, either strongly (P<0.001) or weakly (P=0.09) associated with dominant follicles.

Isolation of genes associated with developmental competency of bovine oocytes.

Eggs differ widely in their ability to develop into an embryo. To address this characteristic, the concept of developmental competency has been coined, defined as the ability or potential of an oocyte to undergo maturation, fertilization and development to blastocyst stages or live offspring. Developmental competency is acquired progressively during folliculogenesis and is linked to follicular size. In an effort to understand the molecular changes underlying differences in competency we compared oocytes derived from large follicles (>or=5mm) to those from small follicles (

Minimally invasive kidney transplantation: the first experience.

Minimally invasive procedures in recent years have gained widespread acceptance. Within the field of transplantation, laparoscopic living donor nephrectomy (LLDN), requiring a 6- to 10-cm incision, is now considered the optimal procedure. According to recent MEDLINE searches, no minimally invasive technique has been reported for kidney transplantation. Considering the rapid evolution of minimally invasive surgery during the last decade, there is little reason to believe that kidney transplantation in future will be excluded from this development. A novel minimally invasive technique for kidney transplantation (MIKT) is presented, restricted to a 7- to 9-cm incision and minimal dissection/tissue trauma. The kidney is meticulously prepared on the back table and placed in a fitted lateral, retroperitoneal pouch. All three anastomoses are performed with the kidney in its final "in situ" position, and ureter reimplantation is done by extravesical technique. Twenty-one patients have been transplanted by MIKT and followed in a prospective manner, along with a matched control group subjected to conventional kidney transplantation. Our results indicate that MIKT may be executed safely and quickly. Beneficial effects on postoperative pain/analgesia, recovery, and complications are suggested by this first MIKT experience. The technical solutions of MIKT are per se not unique. However, the incision is minimal and not larger than the one required for LLDN. Minimally invasive surgery seems particularly attractive in the immunosuppressed population, and even more so with the recent introduction of potent antiproliferative drugs.

On-farm production of AM fungus inoculum in mixtures of compost and vermiculite.

On-farm production of arbuscular mycorrhizal (AM) fungus inoculum can reduce the cost of the inoculum and increase utilization of this symbiosis in plant production. Bahiagrass (Paspalum notatum Flugge) seedlings, colonized by AM fungi, were transplanted into raised bed enclosures. Media within the enclosures was vermiculite mixed with either field soil or yard clippings compost in Experiment I and vermiculite mixed with yard clippings compost or dairy manure/leaf compost in Experiment II. Compost and vermiculite mixtures yielded more propagules of AM fungi than soil-based mixtures in Experiment I. Growth of plants in a 1:4 (v/v) mixture of yard clippings compost and vermiculite produced more inoculum (503 propagules cm(-3)) than growth in 1:9 and 1:99 (v/v) mixtures (240 and 42 propagules cm(-3), respectively). Water, inorganic nutrient solution minus P, and fish protein digest were added to inoculum production enclosures in Experiment II. Results indicated that supplemental nutrient addition was unnecessary. This method produces a concentrated inoculum of AM fungi in a form readily used as an amendment to horticultural potting media for the production of vegetable seedlings.

In vivo 133Cs-NMR a probe for studying subcellular compartmentation and ion uptake in maize root tissue.

Three 133Cs-NMR signals were observed in the spectra of CsCl-perfused and CsCl-grown maize seedling root tips. Two relatively broad lower field resonances were assigned to the subcellular, compartmented Cs+ in the cytoplasm and vacuole, respectively. The rate of area increase of the broader cytoplasmic Cs resonance was about 9-times faster than that of the vacuolar signal during the first 300 min of tissue perfusion with CsCl. In addition, the spin lattice relaxation time of the cytoplasmic Cs resonance was approx. 3-times shorter than that of the extracellular resonance, while the Cs+ signal associated with the metabolically less active vacuolar compartment exhibited a relaxation time comparable to that of the extracellular signal. 133Cs spectra of excised, maize root tips and excised top sections of the root adjacent to the kernel, each grown in 10 mM CsCl showed a difference in the relative areas of the Cs resonance corresponding to the distinct cytoplasm/vacuole volume ratio of these well differentiated sections of the root. The high correlation of counterion concentration with 133Cs chemical shifts suggested that the larger downfield shift exhibited by the cytoplasmic confined Cs+ was due principally to the higher ionic strength and protein content in this compartment. Such observations indicate that 133Cs-NMR might be employed for studying ionic strength, and osmotic pressure associated chemical shifts and the transport properties of Cs+ (perhaps as an analogue for K+) in subcellular compartments of plant tissues.

Structural studies of a phosphocholine substituted beta-(1,3);(1,6) macrocyclic glucan from Bradyrhizobium japonicum USDA 110.

In our previous in vivo 31P study of intact nitrogen-fixing nodules (Rolin, D.B., Boswell, R.T., Sloger, C., Tu, S.I. and Pfeffer, P.E., 1989 Plant Physiol. 89, 1238-1246), we observed an unknown phosphodiester. The compound was also observed in the spectra of isolated bacteroids as well as extracts of the colonizing Bradyrhizobium japonicum USDA 110. In order to characterize the phosphodiester in the present study, we took advantage of the relatively hydrophobic nature of the material and purified it by elution from a C-18 silica reverse-phase chromatography column followed by final separation on an aminopropyl silica HPLC column. Structural characterization of this compound with a molecular weight of 2271 (FAB mass spectrometry), using 13C-1H and 31P-1H heteronuclear 2D COSY and double quantum 2D phase sensitive homonuclear 1H COSY NMR spectra, demonstrated that the molecule contained beta-(1,3); beta-(1,6); beta-(1,3,6) and beta-linked non-reducing terminal glucose units in the ratio of 5:6:1:1, respectively, as well as one C-6 substituted phosphocholine (PC) moiety associated with one group of (1,3) beta-glucose residues. Carbohydrate degradation analysis indicated that this material was a macrocyclic glucan, (absence of a reducing end group) with two separated units containing three consecutively linked beta-(1,3) glucose residues and 6 beta-(1,6) glucose residues. The sequences of beta-(1,3)-linked glucose units contained a single non-reducing, terminal, unsubstituted glucose linked at the C-6 position and a PC group attached primarily to an unsubstituted C-6 position of a beta-(1,3)-linked glucose.

Regional specificity of RAR gamma isoforms in Xenopus development.

All retinoic acid receptors (RAR alpha, beta, gamma) have two isoforms, whose function is unknown. We now show that at least for RAR gamma, the isoforms are differentially distributed in the embryo. RAR gamma 1 and RAR gamma 2 are detected in the head region, whereas RAR gamma 2 is the sole isoform expressed in the tail region. Specifically, it is expressed in the chordoneural hinge, a region of the tailbud that has organizing properties. Treatment with high doses of retinoic acid (RA) reduces expression in this region. The results are discussed in terms of the known teratogenic effects of RA in the tail region.

Partitioning of Intermediary Carbon Metabolism in Vesicular-Arbuscular Mycorrhizal Leek.

Vesicular-arbuscular mycorrhizal fungi are symbionts for a large variety of crop plants; however, the form in which they take up carbon from the host is not established. To trace the course of carbon metabolism, we have used nuclear magnetic resonance spectroscopy with [13C]glucose labeling in vivo and in extracts to examine leek (Allium porrum) roots colonized by Glomus etunicatum (and uncolonized controls) as well as germinating spores. These studies implicate glucose as a likely substrate for vesicular-arbuscular mycorrhizal fungi in the symbiotic state. Root feeding of 0.6 mM 1-[13C]glucose labeled only the fungal metabolites trehalose and glycogen. The time course of this labeling was dependent on the status of the host. Incubation with 50 mM 1-[13C]glucose caused labeling of sucrose (in addition to fungal metabolites) with twice as much labeling in uncolonized plants. There was no detectable scrambling of the label from C1 glucose to the C6 position of glucose moieties in trehalose or glycogen. Labeling of mannitol C1,6 in the colonized root tissue was much less than in axenically germinating spores. Thus, carbohydrate metabolism of host and fungus are significantly altered in the symbiotic state.

Crescent, a novel chick gene encoding a Frizzled-like cysteine-rich domain, is expressed in anterior regions during early embryogenesis.

We describe the isolation of a novel chicken gene that we have termed crescent, based on the most distinctive stage of its highly dynamic expression pattern during early embryogenesis. Crescent encodes a protein that in its N-terminal half shows the characteristic invariant 9 cysteine residues of the cysteine-rich domain (CRD) found in the Frizzled family of proteins, in Smoothened and in Collagen XVIII. The CRD of several Frizzled proteins have recently been shown to bind to Wg. Unlike Frizzled proteins, crescent does not contain a transmembrane domain and thus can not function as a receptor. Crescent expression is first found at stage XII (E-G&K) in the center of the area pellucida. On primitive streak formation, expression is detected in the entire anterior half of the area pellucida in the hypoblast layer. At maximal streak extension, crescent transcripts are localized primarily to the germinal crescent, where the primordial germ cells reside. During head process and head fold stages, crescent labels the anteriormost endodermal cells which will give rise to prospective foregut. With the commencement of somitogenesis, crescent expression rapidly wanes.

Stage- and adult tissue-specific expression of a homeobox gene in embryo and adult Parechinus angulosus sea urchins.

We report the isolation of a gene (PaHbox6), encoding a homeobox-containing protein of the South African sea urchin, Parechinus angulosus. Sequencing identified an Antennapedia-class gene encoding a homeobox that is the homologue of the Hawaiian sea urchin Tripneustes gratilla homeobox gene. Extensive restriction-fragment length polymorphism surrounds the gene. RNase-protection analyses revealed expression of PaHbox6 in mesenchyme blastula embryos at maximal levels of 44 +/- 8 transcripts/embryo. Four adult tissues examined (testes, ovary, intestines, Aristotle's lantern) showed expression of PaHbox6, though at greatly differing levels, with testes highest at eleven transcripts/10 pg RNA. Two transcripts of 5.2 and 5.7 kb were identified in adult tissue.

HLA-restricted cytotoxicity against male-specific (H-Y) antigen after acute rejection of an HLA-identical sibling kidney: clonal distribution of the cytotoxic cells.

A 22-year-old previously nonsensitized female patient (HLA-A2,28; B7,14; DR1,2) received a kidney from her HLA-identical brother. She irreversibly rejected the graft 2 1/2 weeks later. In vitro tests performed 6 weeks after the graft loss, using recipient responding and donor-stimulating and target cells, revealed a weak mixed lymphocyte culture (MLC) response and negative direct cell-mediated lympholysis (CML), but strong cytotoxicity against donor target cells after priming in MLC. Both MLC and CML tests using donor-responding cells and recipient-stimulating and target cells were negative. When the patient's cytotoxic cells were tested on a panel of target cells from nonrelated donors, a significant CML was only found against target cells from male donors sharing HLA-A2 and/or B7 with the patient, not with male target cells sharing A28 or B14. By using cold target inhibition tests, two separate populations of male-specific cytotoxic cells could be demonstrated. The one was restricted by HLA-A2, the other by B7. These findings indicate that a self-HLA-restricted cytotoxicity against a male-specific minor histocompatibility antigen was a major cause of the rejection in this case.

Effects of methylprednisolone on the in vitro generation of human secondary cytotoxic lymphocytes.

The effects of methylprednisolone on the induction of secondary proliferative and cytotoxic lymphocytes in human mixed lymphocyte cultures (MLCs) have been studied. Concentrations of methylprednisolone (MP) as low as 0.01 microgram/ml proved highly effective in inhibiting the generation of cytotoxic memory cells if the steroid was present during the first 5 days of the priming mixed lymphocyte culture. The generation of cytotoxic lymphocytes was also inhibited if the steroid was added along with the restimulating cells on day 10, although the degree of inhibition was not as great as that seen when steroids were added to the cultures at their initiation. Cultures containing 1 microgram of MP per ml during the priming phase required 10 times as many effector cells to achieve a comparable level of cytotoxicity compared to control cultures without steroids. Our results indicate that an important aspect of the immunosuppressive role of steroids is the prevention of the generation of specific memory cells following exposure to alloantigens.