Regulated polyadenylation is a broadly conserved mechanism that controls key events during oogenesis. Pivotal to that mechanism is GLD-2, a catalytic subunit of cytoplasmic poly(A) polymerase (PAP). ...Caenorhabditis elegans GLD-2 forms an active PAP with multiple RNA-binding partners to regulate diverse aspects of germline and early embryonic development. One GLD-2 partner, RNP-8, was previously shown to influence oocyte fate specification. Here we use a genomic approach to identify transcripts selectively associated with both GLD-2 and RNP-8. Among the 335 GLD-2/RNP-8 potential targets, most were annotated as germline mRNAs and many as maternal mRNAs. These targets include gld-2 and rnp-8 themselves, suggesting autoregulation. Removal of either GLD-2 or RNP-8 resulted in shortened poly(A) tails and lowered abundance of four target mRNAs (oma-2, egg-1, pup-2, and tra-2); GLD-2 depletion also lowered the abundance of most GLD-2/RNP-8 putative target mRNAs when assayed on microarrays. Therefore, GLD-2/RNP-8 appears to polyadenylate and stabilize its target mRNAs. We also provide evidence that rnp-8 influences oocyte development; rnp-8 null mutants have more germ cell corpses and fewer oocytes than normal. Furthermore, RNP-8 appears to work synergistically with another GLD-2—binding partner, GLD-3, to ensure normal oogenesis. We propose that the GLD-2/RNP-8 enzyme is a broad-spectrum regulator of the oogenesis program that acts within an RNA regulatory network to specify and produce fully functional oocytes.
In North America, range constraints due to burgeoning development increasingly encroach on wild horse habitat and necessitate effective but humane reproductive management. The largest free-roaming ...wild horse fertility control program by population (>3500) and territory size (≈300,000 acres) is located within Nevada's Virginia Range. Data from a field study investigated porcine zona pellucida (pZP) immunocontraception via remote dart delivery to mares in this population. Analyses aimed to measure efficacy by treatment effects on annual birth rates and population demographics and to evaluate treatment frequency and season against these variables. Analyses included mares' monthly data (January 2019-December 2022; 48 months), characterized by cumulative vaccination numbers subset into four classifications considering the vaccine as having no loss of efficacy or a loss within a 6-, 12-, and 18-month period post vaccination; from foaling data, the likelihood of being in foal and of conceiving in that month; and from age, as mature or immature (<1 years-old). A downward foaling rate and trend in the numbers of mature mares, descriptively presented at monthly intervals, showed markedly declining annual seasonal breeding peaks, with no observed change in foaling season or duration. Within four years, population coverage surpassed 70% and was associated with a 58% reduction in foaling, with only a 10% conception rate. Vaccinated mares increased proportionally: assuming a 12-month decay rate, the system reached stability at an average ≈1.0 vaccination/mare/year, providing a robust recommendation for treatment frequency contributing to best management practices.
Chronic asymptomatic and acute symptomatic anterior uveitis are forms of ocular inflammation associated with juvenileidiopathic arthritis (JIA) Chronic JIA-associated uveitis is characterized by ...young age of onset, female predilection, oligoarthritis,and antinuclear antibody (ANA) positivity. Acute JIA-associated uveitis predominantly affects older male juvenileswho also develop enthesitis. A type I collagen-derived peptide (melanin-associated antigen MAA) induces anterior uveitisin rodents. In this study, we evaluated MAA-induced uveitis in rats as a potential model for JIA-uveitis. We characterizedMAA-induced uveitis by assessing its relationship to age and sex; tracking the occurrence of arthritis, enthesitis, and ANApositivity; and measuring vitreous fluid inflammatory biomarkers. Juvenile and adult and male and female Lewis rats ( Rattusnorvegicus ) were inoculated with MAA. Slit-lamp biomicroscopy, indirect ophthalmoscopy, and joint examinations wereperformed 3 times weekly. Rats were euthanized at 4 wk after MAA inoculation, and plasma ANA testing, vitreous inflammatorybiomarker assays, and globe histopathology assessments were conducted. Uveitis, arthritis, ANA status, levels ofinflammatory biomarkers, histopathology, and joint tomographic images were assessed in relation to age and sex and comparedwith nonuveitic controls. All MAA-immunized rats developed uveitis characterized by anterior chamber fibrin, iridalvessel dilation, and miosis, and uveal and choroidal lymphocytic infiltration. Levels of the vitreous fluid biomarker CCL5were higher in uveitic rats compared with control rats. Time to uveitis onset, clinical uveitis scores, and biomarker levels didnot differ based on age or sex. None of the MAA-exposed rats had arthritis, enthesitis, or ANA. None of the rats inoculatedwith MAA that had been treated with matrix metallopeptidase 1 had clinical, histologic, or immunohistochemical evidenceof ocular inflammation. In contrast to JIA-associated uveitis in humans, MAA-induced uveitis in rats is not associated withage or sex predilections and MAA is not arthritogenic.
Satellite glial cells (SGCs) surrounding primary sensory neurons are similar to astrocytes of the central nervous system in that they buffer the extracellular environment via potassium and calcium ...channels and express the intermediate filament glial fibrillary acidic protein (GFAP). Peripheral nerve injury induces a reactive state in SGCs that includes SGC proliferation, increased SGC/SGC coupling via gap junctions, decreased inward rectifying potassium channel 4.1 (Kir4.1) expression and increased expression of GFAP and the common neurotrophin receptor, p75NTR. In contrast, neuronal p75NTR expression, normally detected in ∼80% of adult rat sensory neurons, decreases in response to peripheral axotomy. Given the differential regulation of p75NTR expression in neurons versus SGCs with injury, we hypothesized that reduced signaling via neuronal p75NTR contributes to the induction of a reactive state in SGCs. We found that reducing neuronal p75NTR protein expression in uninjured sensory neurons by intrathecal subarachnoid infusion of p75NTR‐selective anti‐sense oligodeoxynucleotides for one week was sufficient to induce a “reactive‐like” state in the perineuronal SGCs akin to that normally observed following peripheral nerve injury. This reactive state included significantly increased SGC p75NTR, GFAP and gap junction protein connexin‐43 protein expression, increased numbers of SGCs surrounding individual sensory neurons and decreased SGC Kir4.1 channel expression. Collectively, this supports the tenet that reductions in target‐derived trophic support leading to, or as a consequence of, reduced neuronal p75NTR expression plays a critical role in switching the SGC to a reactive state. GLIA 2014;62:763–777
Main Points:
Sensory neuron p75 neurotrophin receptor (p75NTR) signaling maintains perineuronal satellite glial cell (SGC) homeostatic phenotype.
Reduced p75NTR expression in uninjured sensory neuron induces robust SGC phenotype changes consistent with nerve injury.
For my extended family member, it happened to be about sameness. Since potty training was mostly going to be taught by their father, they wanted their sons to look like him. ...we considered if the ...decision to circumcise should come from him once he became an adult. Whatever the factors could be for an individual or family, the personal decision is always the right choice for you.
Soil carbon (C) storage is generally expressed as a mass of C per unit area measured to a specific depth. This practice is not ideal as the soil surface can fluctuate for several reasons including ...compaction, tillage, and shrink/swell of clays. This change in bulk density may cause the calculated C stocks in a soil to change from one sampling period to the next. In soils with high shrink/swell capacities, the bulk density has the potential to change regardless of soil management. This could present limitations to efforts to monitor changes in C stocks in high shrink/swell soils. Therefore, a study was conducted in two soils with shrink/swell capacity, in which they were sampled to a depth of 90 cm at two or three different moisture conditions and analyzed for bulk density and organic C at 10-cm increments. Profile C stocks were analyzed using the conventional "fixed depth" method and also with a "fixed mass" method. The analysis showed that under moist conditions the swelling of clays did not decrease bulk density. On the contrary, moist conditions generally resulted in elevated bulk density values. This resulted in significant differences in C stocks within depth increments where the variation in C concentration was low. Differences in profile C stocks, as determined using the fixed depth method, caused by differences in bulk density were removed by using the fixed mass method of calculating C stocks. The fixed mass method provided measurements of C stocks, which vary only as a result of variation in C concentrations.
Soil sampling equipment can be a major source of bias in soil organic carbon (SOC) stock estimations. The objective of this study was to evaluate the impact of sampling probes on soil bulk density ...(BD) and SOC stocks calculated using fixed depth (FD) and equivalent soil mass (ESM) methods. Soil samples were collected to 30 cm using three probes with different diameters and divided into 0–10‐, 10–20‐, and 20–30‐cm layers. The probe with smallest diameter measured higher BD at 0–10 cm in 42% of fields and was significantly different when averaged across fields, while no consistent differences were observed at lower depths. This study shows that sampling probes with different diameters may introduce biases in BD and SOC measurements at individual or combined soil layers when calculated using the FD approach. The ESM approach reduced the differences in mean SOC stocks calculated using different probes.
Abstract This study examines the effects of the selective serotonin reuptake inhibitor (SSRI), fluoxetine (PROZAC), on the ontogeny of spontaneous swimming activity (SSA) in developing zebrafish. The ...development of zebrafish motor behavior consists of four sequential locomotor patterns that develop over 1–5 days post fertilization (dpf), with the final pattern, SSA, established at 4–5 dpf. In stage specific experiments, larvae were exposed to 4.6 μM fluoxetine for 24 h periods beginning at 24 h post fertilization (hpf) and extending through 5 dpf. From 1–3 dpf, there was no effect on SSA or earlier stages of motor development, i.e., spontaneous coiling, evoked coiling and burst swimming. Fluoxetine exposure at 3 dpf for 24 h resulted in a transient decrease in SSA through 7 dpf with a complete recovery by 8 dpf. Larvae exposed to 4.6 μM fluoxetine for 24 h on 4 or 5 dpf showed a significant decrease in SSA by day 6 with no recovery through 14 dpf. Although SSA was significantly affected 24 h after fluoxetine exposure, there was little or no effect on pectoral fin movement. These results demonstrate both a stage specific and a long term effect of 4.6 μM fluoxetine exposure in 4 and 5 dpf larvae. Reverse transcriptase polymerase chain reaction (RT-PCR) was performed to determine the relative levels of a serotonin transporter protein (SERT) transcript and the serotonin 1A (5-HT1A ) receptor transcript in developing embryos/larvae over 1–6 dpf. Both transcripts were present at 24 hpf with the relative concentration of SERT transcript showing no change over the developmental time range. The relative concentration of the 5-HT1A receptor transcript, however, showed a two-tiered pattern of concentration. RT-PCR was also used to detect potential changes in the SERT and 5-HT1A receptor transcripts in 6 dpf larvae after a 24 h exposure to 4.6 μM fluoxetine on 5 dpf. Three separate regions of the CNS were individually analyzed, two defined brain regions and spinal cord. The two brain regions showed no effect on transcript levels subsequent to fluoxetine exposure, however, the spinal cord showed a significant decrease in both transcripts. These results suggest a correlation between decreased concentration of SERT and 5-HT1A receptor transcripts in spinal cord and decreased SSA subsequent to fluoxetine exposure.
Background
While intra-arrest echocardiography can be used to guide and monitor chest compression quality, it is not currently feasible on the scene of out-of-hospital cardiac arrests. Rapid and ...automated sonographic localization of the heart may provide first-responders guidance to an optimal area of compression without requiring them to interpret ultrasound images. In this proof-of-concept porcine study, we sought to describe the performance of an automated ultrasound device in correctly identifying and tracing the borders of the heart in three distinct states: pre-arrest, arrest, and late arrest.
Methods
An automated ultrasound device (bladder scanner) was placed on the chests of 7 swine, along the left sternal border (4th–8th intercostal spaces). Scanner-generated images were recorded for each space during pre-arrest, arrest, and finally late arrest. 828 images of the LV and LV outflow tract were randomized and 150 (50/state) selected for analysis. Scanner tracings of the heart were then digitally obscured to facilitate tracing by expert reviewers who were blinded to the physiologic state. Reviewer tracings were compared to bladder scanner tracings; with concordance between these images determined via Sørensen–Dice index (SDI).
Results
When compared to human reviewers, the bladder scanner was able to identify and trace the borders during cardiac arrest. The bladder scanner performed best at the time of arrest (SDI 0.900 ± 0.059). As resuscitation efforts continued and time from initial arrest increased, the scanner’s performance decreased dramatically (SDI 0.597 ± 0.241 in late arrest).
Conclusion
An automated ultrasound device (bladder scanner) reliably traced porcine hearts during cardiac arrest. It is possible a device could be developed to indicate where compressions should be performed without requiring the operator to interpret ultrasound images. Further investigation into rapid, automated, sonographic localization of the heart to identify the area of compression in out-of-hospital cardiac arrest is warranted.
Abstract In this study, p -chlorophenylalanine (pCPA), an inhibitor of tryptophan hydroxylase (the rate limiting enzyme of serotonin synthesis), was used to reduce serotonin (5HT) levels during early ...development in zebrafish embryos. One day old dechorionated embryos were treated with 25 μM pCPA for 24 h and subsequently rescued. Immunohistological studies using a 5HT antibody confirmed that 5HT neurons in the brain and spinal cord were depleted of transmitter by 2 days post fertilization (dpf). Twenty four hours after pCPA exposure embryos were unable to burst swim and were nearly paralyzed. Movement began to improve at 4 dpf, and by 7 dpf, larvae exhibited swimming activity. Rescued larvae continued to grow in rostrocaudal length over 5 days post-rescue, but their length was always 16–21% below controls. Surprisingly, both groups displayed the same number of myotomes. To examine whether hypertonicity of myotomes in treated embryos played a role in their shorter rostrocaudal lengths, 1 dpf embryos were exposed to a combination of 25 μM pCPA and 0.6 mM of the sodium channel blocker ethyl 3-aminobenzoate methanesulfonate (MS-222). After a 24 hour exposure, the embryos exhibited the same rostrocaudal length as control embryos suggesting that myotome hypertonicity plays a major role in the decreased axial length of the treated larvae. In addition, pCPA treated 2 dpf embryos exhibited abnormal notochordal morphology that persisted throughout recovery. Reverse transcriptase polymerase chain reaction (RT-PCR) was performed to determine the relative levels of the serotonin 1A receptor (5HT1A ) transcript and the serotonin transporter (SERT) transcript in the brain and spinal cord of control and treated embryos. Transcripts were present in both brain and spinal cord as early as 1 dpf and reached maximal concentrations by 3 dpf. Embryos treated with pCPA demonstrated a decrease in the concentration of 5HT1A transcript in both brain and spinal cord. While SERT transcript levels remained unaffected in brain, they were decreased in spinal cord. Five days subsequent to pCPA rescue, 5HT1A transcript concentrations remained decreased in brain while SERT transcript levels were elevated in both regions. These findings suggest that reduction of 5HT during early zebrafish development may have an adverse effect on body length, notochordal morphology, locomotor behavior, and serotonin message-related expression.
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