Electrotonic suppression of early afterdepolarizations in isolated rabbit Purkinje myocytes

Many studies suggest that early afterdepolarizations (EADs) arising from Purkinje fibers initiate triggered arrhythmias under pathological conditions. However, electrotonic interactions between Purkinje and ventricular myocytes may either facilitate or suppress EAD formation at the Purkinje-ventricular interface. To determine conditions that facilitated or suppressed EADs during Purkinje-ventricular interactions, we coupled single Purkinje myocytes and aggregates isolated from rabbit hearts to a passive model cell via an electronic circuit with junctional resistance (R(j)). The model cell had input resistance (R(m,v)) of 50 M Omega, capacitance of 39 pF, and a variable rest potential (V(rest,v)). EADs were induced in Purkinje myocytes during superfusion with 1 microM isoproterenol. Coupling at high R(j) to normally polarized V(rest,v) established a repolarizing coupling current during all phases of the Purkinje action potential. This coupling current preferentially suppressed EADs in single cells with mean membrane resistance (R(m,p)) of 297 M Omega, whereas EAD suppression in larger aggregates with mean R(m,p) of 80 M Omega required larger coupling currents. In contrast, coupling to elevated V(rest,v) established a depolarizing coupling current during late phase 2, phase 3, and phase 4 that facilitated EAD formation and induced spontaneous activity in single Purkinje myocytes and aggregates. These results have important implications for arrhythmogenesis in the infarcted heart when reduction of the ventricular mass due to scarring alters the R(m,p)-to-R(m,v) ratio and in the ischemic heart when injury currents are established during coupling between polarized Purkinje myocytes and depolarized ventricular myocytes.     

A retrospective analysis of factors contributing to calf mortality and dystocia in beef cattle

Records of 2191 calvings from the Clemson University Beef Physiology Unit between 1981 and 1993 were analyzed to determine factors affecting malpresentation, mortality and dystocia. Only 20 (0.91%) parturitions involved malpresentation: posterior presentation (n = 14), leg deviations (n = 3), head deviations (n = 2) and breech birth (n = 1). Dystocia affected calf mortality within 24 h of birth (P < 0.05), with mortality increasing as the severity of dystocia increased. There was an overall 4.5% death loss within 24 h of birth, with 4 and 7% mortality rates for calves from multiparous and primiparous dams, respectively (P < 0.05). Mortality was higher for bull vs heifer calves (P < 0.05). Ninety-four percent of calvings were unassisted, while 6% were assisted births. Dystocia was greater (P < 0.01) in primiparous (17%) than in multiparous dams (4%). In births involving dystocia, 28.1% required mild traction, 69.3% required heavy traction and 2.6% required Cesarean section. Birth weights associated with normal births and mild traction (36 and 36 kg) were lighter than those associated with heavy traction and Cesarean section (40 and 42 kg, respectively; P < 0.05). In conclusion, malpresentations were too few to be of significance, and dystocia influenced mortality within 24 h of birth. Calf birth weight and parity of dam explained most of the observed variations in dystocia.     

Comparative effects of early weaning or once-daily suckling on occurrence of postpartum estrus in primiparous beef cows

We concurrently evaluated early weaning and once-daily suckling as options for shortening postpartum interval to estrus in primiparous cows calving in good body condition. Over 2 consecutive years, a total of 77 primiparous cows were assigned to 1 of 3 treatments: control (ad libitum suckling); once-daily suckling (for 30 d prior to start of breeding); or early weaning (calves weaned 30 d prior to start of breeding). All cows had a body condition score (BCS) of > or = 5 at parturition and were maintained in good condition throughout the breeding season. Cows were observed for estrus twice daily and blood samples for subsequent measurement of progesterone were collected at 48-h intervals. For data analysis, cows were divided into early- (> or = 85 d post partum) and late-calving (< 85 d post partum) groups at the onset of the breeding season. All calves were weighed at birth, at the onset of treatment, at weekly intervals until the start of the breeding season, and within 45 d of 205 d of age. Early weaning or once-daily suckling shortened the postpartum interval to first estrus of late-calving primiparous cows by 17 (P < 0.01) and 12 d (P < 0.08), respectively, compared with control group cows. However, among early-calving cows, neither early weaning nor once-daily suckling influenced the postpartum interval. Postpartum intervals of once-daily suckled cows and early-weaned cows were similar (P > 0.10).     

Miniature Swine as a Clinically Relevant Model of Graft-Versus-Host Disease

Miniature swine provide a preclinical model of hematopoietic cell transplantation (HCT) for studies of graft-versus-host disease. HCT between MHC-matched or ‑mismatched pigs can be performed to mimic clinical scenarios with outcomes that closely resemble those observed in human HCT recipients. With myeloablative conditioning, HCT across MHC barriers is typically fatal, with pigs developing severe (grade III or IV) GVHD involving the gastrointestinal tract, liver, and skin. Unlike rodent models, miniature swine provide an opportunity to perform extended longitudinal studies on individual animals, because multiple tissue biopsies can be harvested without the need for euthanasia. In addition, we have developed a swine GVHD scoring system that parallels that used in the human clinical setting. Given the similarities of GVHD in pigs and humans, we hope that the use of this scoring system facilitates clinical and scientific discourse between the laboratory and the clinic. We anticipate that results of swine studies will support the development of new strategies to improve the identification and treatment of GVHD in clinical HCT scenarios.Abbreviations: BMT, bone marrow transplantation; GVHD, graft-versus-host disease; GVL, graft-versus-leukemia; HCT, hematopoietic cell transplantation; TBI, total-body irradiation     

Phylogenetic analysis of the tenascin gene family: evidence of origin early in the chordate lineage

Background  

Tenascins are a family of glycoproteins found primarily in the extracellular matrix of embryos where they help to regulate cell proliferation, adhesion and migration. In order to learn more about their origins and relationships to each other, as well as to clarify the nomenclature used to describe them, the tenascin genes of the urochordate Ciona intestinalis, the pufferfish Tetraodon nigroviridis and Takifugu rubripes and the frog Xenopus tropicalis were identified and their gene organization and predicted protein products compared with the previously characterized tenascins of amniotes.     

Attenuation of pattern recognition receptor signaling is mediated by a MAP kinase kinase kinase

Pattern recognition receptors (PRRs) play a key role in plant and animal innate immunity. PRR binding of their cognate ligand triggers a signaling network and activates an immune response. Activation of PRR signaling must be controlled prior to ligand binding to prevent spurious signaling and immune activation. Flagellin perception in Arabidopsis through FLAGELLIN‐SENSITIVE 2 (FLS2) induces the activation of mitogen‐activated protein kinases (MAPKs) and immunity. However, the precise molecular mechanism that connects activated FLS2 to downstream MAPK cascades remains unknown. Here, we report the identification of a differentially phosphorylated MAP kinase kinase kinase that also interacts with FLS2. Using targeted proteomics and functional analysis, we show that MKKK7 negatively regulates flagellin‐triggered signaling and basal immunity and this requires phosphorylation of MKKK7 on specific serine residues. MKKK7 attenuates MPK6 activity and defense gene expression. Moreover, MKKK7 suppresses the reactive oxygen species burst downstream of FLS2, suggesting that MKKK7‐mediated attenuation of FLS2 signaling occurs through direct modulation of the FLS2 complex.     

Spontaneous calcium transients regulate neuronal plasticity in developing neurons

Calcium ions play critical roles in neuronal differentiation. We have recorded transient, repeated elevations of calcium in embryonic Xenopus spinal neurons over periods of 1 h in vitro and in vivo, confocally imaging fluo 3-loaded cells at 5 s intervals. Calcium spikes and calcium waves are found both in neurons in culture and in the intact spinal cord. Spikes rise rapidly to approximately 400% of baseline fluorescence and have a double exponential decay, whereas waves rise slowly to approximately 200% of baseline fluorescence and decay slowly as well. Imaging of fura 2-loaded neurons indicates that intracellular calcium increases from 50 to 500 nM during spikes. Both spikes and waves are abolished by removal of extracellular calcium. Developmentally, the incidence and frequency of spikes decrease, whereas the incidence and frequency of waves are constant. Spikes are generated by spontaneous calcium-dependent action potentials and also utilize intracellular calcium stores. Waves are produced by a mechanism that does not involve classic voltage-dependent calcium channels. Spikes are required for expression of the transmitter GABA and for potassium channel modulation. Waves in growth cones are likely to regulate neurite extension. The results demonstrate the roles of a novel signaling system in regulating neuronal plasticity, that operates on a time scale 10⁴ times slower than that of action potentials. © 1995 John Wiley & Sons, Inc.