Comparison of individual and combination DNA vaccines for B. anthracis,Ebola virus,Marburg virus and Venezuelan equine encephalitis virus

Multiagent DNA vaccines for highly pathogenic organisms offer an attractive approach for preventing naturally occurring or deliberately introduced diseases. Few animal studies have compared the feasibility of combining unrelated gene vaccines. Here, we demonstrate that DNA vaccines to four dissimilar pathogens that are known biowarfare agents, Bacillus anthracis, Ebola (EBOV), Marburg (MARV), and Venezuelan equine encephalitis virus (VEEV), can elicit protective immunity in relevant animal models. In addition, a combination of all four vaccines is shown to be equally as effective as the individual vaccines for eliciting immune responses in a single animal species. These results demonstrate for the first time the potential of combined DNA vaccines for these agents and point to a possible method of rapid development of multiagent vaccines for disparate pathogens such as those that might be encountered in a biological attack.     

Regulation of hairy-cell survival through constitutive activation of mitogen-activated protein kinase pathways

The hairy cells (HCs) of hairy-cell leukemia are intrinsically activated mature clonal B cells. The aims of this study were to gain further insights into the nature of this activation and to assess its importance for the prolonged HC survival in this chronic disease. We show that HCs contain phosphorylated/activated p38 MAPK, JNK and ERK1/ERK2 (ERK1/2). PKC inhibitors increased the activation of p38 and JNK, but reduced the phosphorylation of ERK1/2. Moreover, PKC inhibition resulted in cell death; cell death was also observed when the activation of ERK1/2 in HCs was abrogated with an inhibitor of MEK1/2 activation. In addition to PKC, active Src kinase was also shown to be involved in the maintenance of Raf-independent ERK activation in HCs. During cell culture on a nonadherent surface, ERK phosphorylation was sustained, while phosphorylation of p38 and JNK decreased. This decrease was not observed in HCs cultured on vitronectin (VN), indicating that p38/JNK activation is probably a consequence of in vivo HC interaction with VN present in abundance in the red pulp of the spleen. Taken together, these results suggest that active p38/JNK make HCs susceptible to apoptosis, but the cells are effectively rescued by ERK activation involving constitutively active PKC and Src. These findings are relevant for the understanding of the prolonged cell survival of HCs and their selective sensitivity to some chemotherapeutic agents.     

Effects of mistletoe lectin I and ionizing radiation on the glucose and thymidine uptake in tumour cells in vitro

The increased uptake of hexose by mammalian cells is considered to be a general response to stress. Nowadays, mistletoe lectin separated from the extracts of the European mistletoe (Viscum album L.) is often used in adjuvant cancer therapy. The present work studies the effect of the lectin on unirradiated and x-irradiated tumour cells. The response of cultured human lung carcinoma cells (Calu-1) was followed by radioactive glucose uptake as well as by tritiated thymidine incorporation. The cells were maintained either in a complete or a so-called restrictive medium. Slight metabolic changes were found in the restrictive medium but not in the complete one. Mistletoe lectin I at a very low concentration (0.001 ng/mL) increased the glucose uptake and thymidine incorporation. Ionizing radiation (1 Gy) did not influence the hexose uptake but it enhanced the incorporation of thymidine. It seems that the actions of two different factors (mistletoe lectin I and radiation) proved to be rather provoking stress effects for the tumour cells as detected in the restrictive medium.     

Outcome in congenital stridor (laryngomalacia)

Twenty-one children who had been diagnosed as having laryngomalacia by direct laryngoscopy in infancy were reviewed 7 to 12 years later. The natural history of the disease is documented. A wide variation in the time of onset and duration of the stridor was found and there was a high incidence of feeding difficulties. A previously reported association with mental retardation or cerebral palsy is not confirmed. 4 out of the 21 children had early speech problems.     

Precise intraoperative location of gastrointestinal bleeding with a hand-held counter. Work in progress

The nuclear medicine bleeding scan is frequently insufficient to locate sites of bleeding precisely, in spite of its great sensitivity. A small, hand-held Geiger-Müller counter, placed directly on exposed intestine in the operating room, enables precise location of the probable bleeding site. In three patients, the technique allowed a minimal amount of intestine to be resected, distinguished between large- and small-intestinal hemorrhage, and eliminated other foci as sites of bleeding.     

Glycinergic and GABAergic calcium responses in the developing lateral superior olive

The lateral superior olive (LSO), a binaural nucleus involved in sound localization, receives tonotopically organized inhibitory inputs from the medial nucleus of the trapezoid body (MNTB). During development, the tonotopic organization of this glycinergic/GABAergic MNTB-LSO pathway is established by activity-dependent axonal reorganization. However, the underlying mechanisms by which this reorganization takes place have remained largely unknown. As cytosolic calcium is one of the most important second messengers responsible for inducing synaptic plasticity and reorganization, we examined whether and how activity in the MNTB-LSO pathway changes the intracellular calcium concentration ([Ca2+]i) in developing LSO neurons. By applying calcium imaging techniques to Fura-2-labelled slices from neonatal rats and mice, we found that glycine and GABA (gamma-aminobutyric acid) affect [Ca2+]i in LSO neurons in an age-dependent manner; during the first postnatal week, the period at which glycine and GABA are depolarizing in the LSO, glycine and GABA always increased [Ca2+]i. However, in 2-week-old animals, the time around hearing onset when glycine and GABA are hyperpolarizing, glycine and GABA slightly decreased [Ca2+]i. Calcium responses could also be elicited by stimulation of afferent fibres from the MNTB, and these synaptic responses were mediated by glycine and GABA(A) receptors. Furthermore, GABA, which is a neurotransmitter only in the immature MNTB-LSO pathway, played a major role in generating MNTB-elicited Ca2+ responses. The direct link of glycinergic/GABAergic synaptic activity to intracellular calcium signalling during the period of inhibitory synaptic plasticity could be one of the mechanisms by which tonotopic MNTB-LSO connections become established.     

PLACENTAL CALCIUM PUMP: CLINICAL-BASED EVIDENCE

Placenta can be considered as a pump of calcium necessary for the normal development of the fetus. We believe that the location of this pump is in the placental basement membrane. The calcification of this membrane has been described only in cases of in utero fetal death. In this study we describe for the first time a case of placental calcification in a living fetus. The fetus of a normal 21-year-old pregnant woman showed heart abnormalities but the genetic analysis showed a normal male karyotype. The histology of the placenta demonstrated multiple intravillous linear and granular calcific incrustations The hemtoxylin/eosin stain of the sections revealed basement membrane calcific incrustations and intravillous calcium deposits. We postulate that the fetal circulation in the villi was impaired and the calcium that reached the villi from the mother was deposited at this level.     

Successes and Challenges of Interprofessional Physiologic Birth and Obstetric Emergency Simulations in a Nurse‐Midwifery Education Program

This article describes childbirth simulation design and implementation within the nurse‐midwifery education program at the University of California, San Francisco. Nurse‐midwife and obstetrician faculty coordinators were supported by faculty from multiple professions and specialties in curriculum review and simulation development and implementation. The primary goal of the resulting technology‐enhanced simulations of normal physiologic birth and obstetric emergencies was to assist learners’ development of interprofessional competencies related to communication, teamwork, and patient‐centered care. Trainees included nurse‐midwifery students; residents in obstetrics, pediatrics, and family medicine; medical students; and advanced practice nursing students in pediatrics. The diversity of participant types and learning levels provided benefits and presented challenges to effective scenario‐based simulation design among numerous other theoretical and logistical considerations. This project revealed practical solutions informed by emerging health sciences and education research literature, faculty experience, and formal course evaluations by learners. Best practices in simulation development and implementation were incorporated, including curriculum revision grounded in needs assessment, case‐ and event‐based clinical scenarios, optimization of fidelity, and ample time for participant debriefing. Adequate preparation and attention to detail increased the immersive experience and benefits of simulation. Suggestions for fidelity enhancement are provided with examples of simulation scenarios, a timeline for preparations, and discussion topics to facilitate meaningful learning by maternity and newborn care providers and trainees in clinical and academic settings. Pre‐ and postsimulation measurements of knowledge, skills, and attitudes are ongoing and not reported. This article is part of a special series of articles that address midwifery innovations in clinical practice, education, interprofessional collaboration, health policy, and global health.