Equine non-invasive time-lapse imaging and blastocyst development.

In this study we examined the timeline of mitotic events of invitro-produced equine embryos that progressed to blastocyst stage using non-invasive time-lapse microscopy (TLM). Intracytoplasmic sperm injection (ICSI) embryos were cultured using a self-contained imaging incubator system (Miri®TL; Esco Technologies) that captured brightfield images at 5-min intervals that were then generated into video for retrospective analysis. For all embryos that progressed to the blastocyst stage, the initial event of extrusion of acellular debris preceded all first cleavages and occurred at mean (±s.e.m.) time of 20.0±1.1h after ICSI, whereas 19 of 24 embryos that did not reach the blastocyst stage demonstrated debris extrusion that occurred at 23.8±1.1h, on average 4h longer for this initial premitotic event (P<0.05). Embryos that failed to reach the blastocyst stage demonstrated a 4-h delay compared with those that reached the blastocyst stage to reach the 2-cell stage (P<0.05). All embryos that reached the blastocyst stage expressed pulsation of the blastocyst with visible expansion and contraction at approximate 10-min intervals, or five to six times per hour. Using a logit probability method, we determined that 2- and 8-cell stage embryos could reasonably predict which embryos progressed to the blastocyst stage. Together, the results indicate that TLM for equine embryo development is a dynamic tool with promise for predicting successful embryo development.

Pharmacological enhancement of mGlu5 receptors rescues behavioral deficits in SHANK3 knock-out mice.

SHANK3 (also called PROSAP2) genetic haploinsufficiency is thought to be the major cause of neuropsychiatric symptoms in Phelan-McDermid syndrome (PMS). PMS is a rare genetic disorder that causes a severe form of intellectual disability (ID), expressive language delays and other autistic features. Furthermore, a significant number of SHANK3 mutations have been identified in patients with autism spectrum disorders (ASD), and SHANK3 truncating mutations are associated with moderate to profound ID. The Shank3 protein is a scaffold protein that is located in the postsynaptic density (PSD) of excitatory synapses and is crucial for synapse development and plasticity. In this study, we investigated the molecular mechanisms associated with the ASD-like behaviors observed in Shank3Δ11-/- mice, in which exon 11 has been deleted. Our results indicate that Shank3 is essential to mediating metabotropic glutamate receptor 5 (mGlu5)-receptor signaling by recruiting Homer1b/c to the PSD, specifically in the striatum and cortex. Moreover, augmenting mGlu5-receptor activity by administering 3-Cyano-N-(1,3-diphenyl-1H-pyrazol-5-yl)benzamide ameliorated the functional and behavioral defects that were observed in Shank3Δ11-/- mice, suggesting that pharmaceutical treatments that increase mGlu5 activity may represent a new approach for treating patients that are affected by PMS and SHANK3 mutations.

Performance evaluation of the Abbott CELL-DYN Emerald for use as a bench-top analyzer in a research setting.

INTRODUCTION: The CELL-DYN Emerald is a compact bench-top hematology analyzer that can be used for a three-part white cell differential analysis. To determine its utility for analysis of human and mouse samples, we evaluated this machine against the larger CELL-DYN Sapphire and Sysmex XT2000iV hematology analyzers. METHODS: 120 human (normal and abnormal) and 30 mouse (normal and abnormal) samples were analyzed on both the CELL-DYN Emerald and CELL-DYN Sapphire or Sysmex XT2000iV analyzers. For mouse samples, the CELL-DYN Emerald analyzer required manual recalibration based on the histogram populations. RESULTS: Analysis of the CELL-DYN Emerald showed excellent precision, within accepted ranges (white cell count CV% = 2.09%; hemoglobin CV% = 1.68%; platelets CV% = 4.13%). Linearity was excellent (R² ≥ 0.99), carryover was minimal (<1%), and overall interinstrument agreement was acceptable for both human and mouse samples. Comparison between the CELL-DYN Emerald and Sapphire analyzers for human samples or Sysmex XT2000iV analyzer for mouse samples showed excellent correlation for all parameters. CONCLUSION: The CELL-DYN Emerald was generally comparable to the larger reference analyzer for both human and mouse samples. It would be suitable for use in satellite research laboratories or as a backup system in larger laboratories.

Effects of season and distance moved during loading on transport losses of market-weight pigs in two commercially available types of trailer.

This study evaluated effects of trailer design and season on physical indicators of stress during loading and unloading and transport losses (dead and nonambulatory pigs) in market-weight pigs (BW = 129.6 +/- 0.40 kg). A total of 109 trailer loads of pigs (n = 17,256 pigs) from 1 farm were used in a randomized complete block design with a 2 x 4 factorial arrangement of treatments: 1) trailer design (potbelly vs. straight-deck) and 2) season (spring vs. summer vs. fall vs. winter). A subset of loads (n = 42) was used to examine effect of distance pigs were moved during loading [short (<24 m) vs. long (47 to 67 m)] on physical indicators of stress and transport losses. This study was conducted on 7 d per season at 1 farm with 4 loads (2 on potbelly and 2 on straight-deck trailers) being transported each day to 1 commercial packing plant. Pigs from different farm groups were mixed on the trailer and provided with 0.45 m(2)/pig floor space during an approximately 4-h journey to the plant. The percentage of pigs exhibiting open-mouth breathing, skin discoloration, and muscle tremors was recorded during loading and unloading. Additionally, dead pigs on arrival at the plant and nonambulatory pigs at the farm and at the plant were recorded. Effects of trailer design on open-mouth breathing and skin discoloration during unloading were dependent on season (trailer design x season interaction; P < 0.05). Pigs unloaded from potbelly trailers had a greater (P < or = 0.05) incidence of open-mouth breathing in the spring and summer and a greater (P < 0.05) incidence of skin discoloration in the spring, summer, and winter than pigs unloaded from straight-deck trailers. The incidence of total nonambulatory pigs at the plant was greater (P < 0.05) in the winter than in the spring and summer. The long compared with short distance moved treatment resulted in a greater (P = 0.001) incidence of open-mouth breathing and skin discoloration during loading and tended (P = 0.06) to increase the incidence of nonambulatory pigs at the farm. However, there was no effect of trailer design, season, or loading distance on total losses at the plant. In summary, physical indicators of stress (open-mouth breathing and skin discoloration) were increased with the long distance moved during loading treatment and were greater during unloading for potbelly than straight-deck trailers; however, trailer design, season, and loading distance had minimal effects on total transport losses.

Frequency of the HAL-1843 mutation of the ryanodine receptor gene in dead and nonambulatory-noninjured pigs on arrival at the packing plant.

Four Midwestern packing plants (designated as plants A, B, C, and D) were visited on 53 occasions, and tissue samples were collected postmortem from a total of 2,019 pigs to determine the frequency of the HAL-1843 mutation of the ryanodine receptor gene in dead (DOA), nonambulatory-noninjured (NANI), and normal animals. The sampled pigs came from approximately 130,000 animals from 454 farms and were transported on 861 trailer loads, with an average of 152 pigs/load and an average pig live BW/load of 125 (SD 7.02) kg/pig. Frequency of animals with the HAL-1843 mutation was low, with only 2.7% of the pigs being either homozygous recessives (nn, 0.45%) or carriers (Nn, 2.3%) for the mutation and 97.3% of the pigs being homozygous for the normal allele (NN). The mutation was present in all 3 classes of pig, with 1.8% of normal, 1.8% of NANI, and 4.7% of DOA animals having at least 1 copy. Two of the plants (A and C) had a greater frequency (P < 0.05) of carrier (3.7 and 3.5 vs. 1.1 and 1.0 for plants A and C vs. B and D, respectively) and homozygous recessive (1.0 and 0.9 vs. 0.0 and 0.0, respectively) animals than the others (plants B and D). There was a greater frequency (P < 0.05) of carriers in DOA animals than in the normal or NANI pigs (3.7 vs. 1.7 and 1.5 for DOA vs. normal and NANI, respectively). The 55 pigs that had at least 1 copy of the mutation came from 53 farms; therefore, the mutation was relatively widespread, being present in approximately 11% of the farms sampled. Although the HAL-1843 mutation is still present in commercial pig populations in the United States, its low frequency in DOA and NANI pigs suggests that it is not a major cause of these transport losses.

Role of the JAKs/STATs pathway in the intracellular calcium changes induced by interleukin-6 in hippocampal neurons.

Recent studies show that inflammation has an active role in the onset of neurodegenerative diseases. It is known that in response to extracellular insults microglia and/or astrocytes produce inflammatory agents. These contribute to the neuropathological events in the aging process and neuronal degeneration. Interleukin-6 (IL-6) has been involved in the pathogenesis of neurodegenerative disorders, such as Alzheimer's and Parkinson's diseases. Here, we show that IL-6 treatment of rat hippocampal neurons increases the calcium influx via NMDA-receptor, an effect that is prevented by the specific NMDA receptor antagonist MK-801 (dizocilpine). We also show that this calcium influx is mediated by the JAKs/STATs pathway, since the inhibitor of JAKs/STATs pathway, JAK 3 inhibitor, blocks calcium influx even in the presence of IL-6. This increase in calcium signal was dependent on external sources, since this signal was not observed in the presence of EGTA. Additional studies indicate that the increase in cytosolic calcium induces tau protein hyperphosphorylation, as revealed by using specific antibodies against Alzheimer phosphoepitopes. This anomalous tau hyperphosphorylation was dependent on both the JAKs/STATs pathway and NMDA receptor. These results suggest that IL-6 would induce a cascade of molecular events that produce a calcium influx through NMDA receptors, mediated by the JAKs/STATs pathway, which subsequently modifies the tau hyperphosphorylation patterns.

Coledocoduodenostomía laterolateral tipo Jurasz / Jurasz type lateral lateral coledochoduodenostomy

Se analiza una serie de 21 pacientes operados entre junio de 1989 y junio de 1993 por patología biliar, a quienes se realizó coledocoduodenostomía tipo Jurasz, por indicaciones como: estenosis de colédoco distal, panlitiasis, cálculo impactado, con vía biliar dilatada más de 16 mm. El 71 por ciento de los pacientes presentaba colangitis al momento de la derivación. Se demostró en el seguimiento endoscópico gastritis en 40 por ciento de estos pacientes sometidos a biopsia, y en 20 de ellos se demostró atipias de mucosa gástrica secundaria sugerentemente a reflujo de bilis. No se observó colangitis aguda postoperatorias, ni litiasis recurrente. Todos los pacientes han experimentado notoria mejoría postderivación, con un bajo índice de complicaciones postcirugía