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Electrical stimulation of the hippocampus and acquisition of the conditioned avoidance response in shuttle-box in cats.

The effects of electrical stimulation and electrolytic lesions of the posterior part of the hippocampus on acquisition of the active avoidance response (AAR) in shuttle-box were studied in 35 cats. Both weak or strong hippocampal stimulation applied simultaneously with conditioned stimulus (CS) facilitated the acquisition of AAR. After discontinuation of stimulation the level of AAR performance in response to CS alone decreased in cats trained with the use of strong stimulation. However, if the stimulation was applied before each experimental session the level of AAR performance did not differ from that observed during the final phase of training. These changes of AAR were not observed in cats trained with the use of weak stimulation. The application of hippocampal stimulation alone did not evoke AAR in shuttle-box after training independently of stimulation parameters. There were no differences in the speed of AAR acquisition between unoperated cats, implanted controls and cats with lesions in the posterior part of the hippocampus.

Animals

A cold box for the transport and storage of vaccines.

A cold box capable of maintaining a temperature below +4 degrees C for 1 week was constructed and tested in the laboratory and under field conditions. Cooling is produced by commercial cold packs precooled in a deep-freeze or the freezing compartment of a refrigerator. The box can take approximately 3000 doses of vaccine and is simple, cheap and strong. It is primarily intended for storage of vaccines during field trips by vaccination teams, as an alternative to the refrigerator in regional and peripheral stores in the case of an electrical power failure, and for the delivery of vaccines from regional store to the district.

Cold Temperature

Continuous airway pressure breathing with the head-box in the newborn lamb: effects of regional blood flows.

Continuous airway pressure delivered by a head-box is an accepted means of treating clinical hyaline membrane disease. To investigate hemodynamic alterations resulting from its use, eight newborn lambs, 1 to 6 days of age, were studied at 6 and 11 mm Hg of positive pressure, while spontaneoulsy breathing room air. Organ blood flows and cardiac output were measured with 25 micron-diameter radioactive microspheres. Heart rate, left ventricular pressure, and arterial blood gases did not change during the study. Jugular venous pressures increased from 6.4 mm Hg to 18.6 and 24.2 mm Hg at 6 and 11 mm Hg, respectively (P less than .005). Cardiac output decreased approximately 20% at either intrachamber pressure setting. Renal blood flow fell 21% at 11 mm Hg. No significant changes in blood flow were found in the brain, gastrointestinal tract, spleen, heart, or liver when compared to control flows. Of particular interest was the finding of a 28% reduction in ocular blood flow at 6 mm Hg and 52% at 11 mm Hg. From these results, we conclude that substantial cardiovascular alterations may occur during the application of head-box continuous airway pressure breathing, including a significant reduction in ocular blood flow.

Animals

[Sterilization of plastic culture boxes by formaldehyde vapour (author's transl)].

Various experiments showed that plastic culture boxes may be sterilized by formaldehyde vapour in an exsiccator. Detectable quanties of formaldehyde were no longer found to be present when the dishes had been stored at room temperature for 24 hours after sterilization. A striking fature in these experiments consisted in the fact that colonies failed to grow on those agar plates, the surfaces of which had been in contact with formaldehyde vapour. The dishes used should be meticulously cleaned as studies showed that complete sterilization was not achieved when bacteria were present in or on agar. When Petri dishes cleansed in alcohol and then sterilized were used in assessing aerobic bacterial counts in minced meat by the mixed plate technique, significantly higher average numbers of colonies were recorded than was the case when commercially available sterilized Petri dishes were used. When the streak plate method was adopted, however, no significant differences were observed. When all the required manoeuvres are performed with meticulous care, the method is a practicable one ensuring that plastic culture boxes may be used again several times.

Formaldehyde

Plant U-box E3 ligases: Versatile regulators of environmental stress adaptation and ABA signaling.

Ubiquitination is a reversible post-translational modification that orchestrates a wide spectrum of fundamental processes throughout the plant life cycle. Executed by a hierarchical E1-E2-E3 cascades, this modification tags targets with ubiquitin to modulate their turnover, activity, or subcellular compartmentalization. Among the diverse E3 ligase families, plant U-box (PUB) proteins stand out as a prominent class that determines substrate selection and has emerged as a focal point of stress biology. In this review, we first delineate the structural features of PUB proteins, highlighting their conserved domains and associated regulatory motifs. We then systematically dissect their multifaceted functions in abiotic stress adaptation, encompassing drought, salinity, extreme temperatures, oxidative stress, heavy metal toxicity, with particular emphasis on their integration with ABA signaling networks. We further outline critical knowledge gaps and propose future strategies to decode the regulatory architecture of PUBs. Collectively, this review provides a theoretical foundation and new insights for facilitating the genetic improvement of crop resilience in the face of continuously intensifying environmental stresses through the manipulation of PUB-mediated ubiquitination networks.

ABA signaling

Oncogenic DEAD-box ATPase DDX41 establishes transcript ensembles via CLK3-dependent and -independent mechanisms.

Post-transcriptional diversification of RNA transcripts mediated by complex processing machinery, including DEAD-box ATPases, establishes and maintains cellular phenotypes. For example, DDX41 controls RNA splicing, innate immune signaling, and genome stability. Although heterozygous DDX41 germline genetic variation occurs in familial myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML), the DDX41 contributions to splicing globally, biological processes, and pathogenic mechanisms are incompletely defined. Using a genetic rescue system with Ddx41+/- myeloid progenitors, we established global wildtype DDX41 and pathogenic variant mechanisms. Differing from pathogenic variants of other RNA splicing regulators, DDX41 deficiency compromised multiple splicing steps. DDX41-regulated transcripts encoded factors controlling RNA splicing, including Cdc2-like kinase 3 (CLK3). DDX41 regulated Clk3 transcripts, and elevated CLK3 during myeloid differentiation. Loss-of-function analysis revealed DDX41-regulated splicing commonly, but not always, required CLK3. Thus, through a mechanism utilizing a splicing factor kinase that itself is DDX41-regulated, DDX41 establishes transcript ensembles in myeloid progenitors.

DEAD-box RNA Helicases

Euflavine: effect on brain heavy metal content and staining pattern, and on shuttle box behavior in goldfish.

Goldfish were exposed to euflavine in doses of 7-100 mg/l for 1h-14 days. The treatment resulted in a reduced stainability of the brain with the Timm histochemical procedure for "heavy metals". In contrast, analysis by flameless atomic absorption spectrophotometry showed no significant changes in the concentrations of zinc and of copper after euflavine treatment. In the shuttle box a significant increase in general activity was found. These results, compared to earlier work with other chelating agents, indicate that such drugs do not have a uniform effect on behavior, and that they do not cause a true depletion of heavy metals in the brain.

Acridines

Morphine and shuttle-box self-stimulation in the rat: a model for euphoria.

In a shuttle-box self-stimulation paradigm, analgesic doses of morphine increase the amount of time a rat leaves rewarding brain stimulation on, without altering average OFF times. This paradigm may serve as a model for the euphoria induced by narcotic drugs and as a useful tool for evaluating the reinforcing effects of drugs.

Animals

Tolerance to the effects of delta9-THC on shuttle-box performance and body temperature.

Two groups of rats were trained in a shuttle-box and received delta9-tetrahydrocannabinol (delta9-THC), either before or after being tested. The drug-before group showed tolerance--within 3-6 sessions--to the response-inhibiting effect of THC. The drug-after animals appeared also to be tolerant when they received delta9-THC before being tested. It is concluded that the tolerance to this effect probably is not learned, but has a physiological base. This is corroborated by the finding that during the same study all the animals developed tolerance to the hypothermic effect of delta9-THC.

Animals

Etorphine and shuttle-box self-stimulation in the rat.

Rats were trained to turn rewarding electrical brain stimulation on and off by crossing back and forth in a shuttle-box. Moderate doses of the narcotic analgesic, etorphine, increased mean ON times while having little effect on OFF times. Tolerance did not develop to the reward enhancement action over five consecutive days of injections. This paradigm seems valuable for exploration of the reinforcing properties of narcotic drugs.

Animals

Evolutionary architecture and lineage-specific diversification of Forkhead box transcription factors in Perna viridis.

The Forkhead box (Fox) transcription factors are evolutionarily conserved regulators of development, cell cycle, and apoptosis across metazoans. This study provides the first comprehensive genome-wide analysis of the Fox gene family in the Asian green mussel (Perna viridis). We identified 28 Fox genes distributed across 10 chromosomes. Comparative analysis reveals the absence of the FoxI, FoxQ1, FoxR and FoxS subfamily, consistent with other bivalves and indicative of lineage-specific gene loss during molluscan evolution. Notably, gene duplications in the FoxAB, FoxD, FoxH, FoxN1-4, FoxQ2 and FoxQD subfamilies may reflect functional diversification associated with environmental adaptation. Exon-intron structural variability, including intron loss in several paralogues, suggests structural diversification and potential regulatory variation. Phylogenetic reconstruction confirmed the monophyly of core Fox classes while highlighting divergent expansion patterns in lophotrochozoans. Selection analyses showed strong purifying selection across duplicated Fox paralogs, supporting functional conservation after lineage-specific expansion. Gene Ontology enrichment linked Fox genes to stress response, apoptosis, and transcriptional regulation. By integrating phylogenetic, structural, and transcriptomic analyses, this study provides a genomic framework for understanding Fox gene organisation, evolution, and tissue-associated expression patterns in Perna viridis and establishes a comparative resource for future functional studies in bivalves.

Animals

A wild soybean MADS-box gene GsAGL62 improves seed weight by enhancing cytokinin signaling and cell proliferation.

Soybean seed weight is a key yield determinant, but the transcriptional mechanisms connecting hormone signaling to seed growth are poorly understood. Here, we identify GsAGL62, a wild soybean MADS-box transcription factor located within a previously mapped hundred-seed weight (HSW) locus and a domestication-associated selective sweep. Functional analyses show that overexpression of GsAGL62 in cultivated soybean significantly increases HSW, whereas ethyl methanesulfonate (EMS)-induced gmagl62 mutants reduce it. Integrated transcriptomic and metabolomic analyses reveal that GsAGL62 enhances cytokinin accumulation and signaling cytokinin-associated responses, accompanied by increased expression of genes involved in cell proliferation. Mechanistically, GsAGL62 directly binds to the promoter of the conserved growth inhibitor GmATPK2 and represses its transcription. Consistently, independent EMS-induced gmatpk2 mutants exhibit increased seed weight, supporting GmATPK2 as a downstream negative regulator of seed growth. Population genetic analyses further reveal strong differentiation of GsAGL62 promoter haplotypes during soybean domestication and improvement. These haplotypes show differential promoter activities and are associated with distinct agronomic performance, suggesting that cis-regulatory variation at GsAGL62 contributes to its selection during soybean improvement. Collectively, our findings establish a regulatory module linking GsAGL62 to cytokinin-associated responses, cell proliferation, and seed growth, and highlight GsAGL62 as a potential target for soybean yield improvement.

Cell proliferation

Stunkardia minuta sp. n. (Trematoda: Paramphistomidae) from the Malayan box-tortoise Cuora amboinensis.

Stunkardia minuta sp. n. was recovered from the small intestine and rectum of 5 box-tortoises (Cuora amboinensis) in Malaysia. The average body size (L X W) is 11-42 X 2-35 mm; oral sucker 1-51 X 1.02; oral pouches 0-21 X 0.18; ventral sucker 1-67 X 1-43; oesophageal bulb 0-54 X 0-54; ant. testis 0-95 X 0-98; post. testis 0-94 X 0.94; seminal vesicle 0-82 X 0-24; ootype 0-21 X 0-41; ovary 0-41 X 0-34; and egg 121 X 83 micrometer. Although morphologically similar to S. dilymphosa, S. minuta is distinct from any other reported member of the Paramphistomidae.

Animals

Hormonal dissociation of limbic lesion effects on shuttle box avoidance in rats.

Rats with septal or hippocampal lesions, relative to normal rats, showed facilitated acquisition of a shuttle box avoidance response. The rats with septal lesions were also highly resistant to extinction compared with normal rats. When the same lesion effects were examined in hypophysectiomized rats, the animals with septal lesions continued to show facilitated performance, and those rats with hippocampal lesions performed no differently than nonoperated control animals. These findings are consistent with the hypothesis that the facilitated avoidance performance found in rats with hippocampal lesions is attributable to lesion-induced changes in hypophyseal activity, but similar changes induced by septal lesions are not.

Acoustic Stimulation

Expanding the amino acid repertoire of ribosomal polypeptide synthesis via the artificial division of codon boxes.

In ribosomal polypeptide synthesis the library of amino acid building blocks is limited by the manner in which codons are used. Of the proteinogenic amino acids, 18 are coded for by multiple codons and therefore many of the 61 sense codons can be considered redundant. Here we report a method to reduce the redundancy of codons by artificially dividing codon boxes to create vacant codons that can then be reassigned to non-proteinogenic amino acids and thereby expand the library of genetically encoded amino acids. To achieve this, we reconstituted a cell-free translation system with 32 in vitro transcripts of transfer RNASNN (tRNASNN) (S = G or C), assigning the initiator and 20 elongator amino acids. Reassignment of three redundant codons was achieved by replacing redundant tRNASNNs with tRNASNNs pre-charged with non-proteinogenic amino acids. As a demonstration, we expressed a 32-mer linear peptide that consists of 20 proteinogenic and three non-proteinogenic amino acids, and a 14-mer macrocyclic peptide that contains more than four non-proteinogenic amino acids.

Amino Acid Sequence

Telomere-to-telomere genome of Phoebe chekiangensis reveals that age-dependent CHG hypomethylation promotes floral transition via MADS-box gene activation.

Phoebe species are renowned for their highly valuable 'golden thread' timber; however, their protracted juvenile phase presents a significant obstacle to mechanistic investigations of floral induction. Phoebe chekiangensis, a rare early-flowering representative within this genus, provides a unique model system for dissecting the vegetative-to-reproductive phase transition. Nevertheless, the absence of a high-quality reference genome has severely hindered molecular insights into its developmental regulation. Here, we present the first telomere-to-telomere (T2T) genome assembly for P. chekiangensis, comprising two completely gap-free haplotypes with contig N50 values exceeding 65 Mb, base-level quality scores >36, and Long Terminal Repeat Assembly Index scores surpassing the gold standard threshold of 20. Approximately 29 000 genes were annotated per haplotype, supported by a BUSCO completeness score of >97%. Age-resolved transcriptomic landscapes identified two MADS-box transcription factors, PcMADS5 (AP1-like) and PcMADS19.1 (SOC1-like), as core activators of the floral transition. Both genes triggered precocious flowering when ectopically expressed in Arabidopsis thaliana. Whole-genome bisulfite sequencing revealed a progressive, age-dependent decline in CHG (where H is A, C, or T) DNA methylation, which was particularly pronounced at the PcMADS19.1 locus. Notably, DML1/2, which mediate active DNA demethylation, were coordinately upregulated during the onset of reproductive growth. Chemical demethylation using 5-azacytidine further diminished CHG methylation and selectively enhanced PcMADS19.1 expression, confirming a causal relationship between CHG hypomethylation and transcriptional activation. This work delivers the first chromosome-scale T2T genome within the genus Phoebe and uncovers CHG demethylation as a previously unrecognized epigenetic switch governing reproductive competence in woody perennials.

Journal Article

Evaporative water loss in box turtles: effects of rostral brainstem and other temperatures.

Box turtles were implanted with thermodes astride the preoptic tissue of the brainstem. The rate of evaporative water loss could be transiently increased by heating the rostral brainstem. Heating tissue in the anterior hypothalamus affected evaporative water loss only a high ambient temperatures. The magnitude of the response was proportional both to the change in hypothalamic temperature and to the ambient temperature with which the turtle was in equilibrium. The major fuction of a high rate of evaporative water loss in turtles is probably to protect the brain from overheating during thermal stress.

Animals

Rapid entry port for an anaerobic glove box.

A rapid entry mechanism for an anaerobic glove box is described. The entry port is practical for entry or removal of one or two items and requires only seconds to operate. No deleterious effect on isolation or growth of anaerobes has been encountered since installation of the rapid entry port 3.5 years ago. Details for its construction are given.

Anaerobiosis