Artificial intelligence-based morphologic classification and molecular characterization of neuroblastic tumors from digital histopathology.

A deep learning model using attention-based multiple instance learning (aMIL) and self-supervised learning (SSL) was developed to perform pathologic classification of neuroblastic tumors and assess MYCN-amplification status using H&E-stained whole slide digital images. The model demonstrated strong performance in identifying diagnostic category, grade, mitosis-karyorrhexis index (MKI), and MYCN-amplification on an external test dataset. This AI-based approach establishes a valuable tool for automating diagnosis and precise classification of neuroblastoma tumors.

Serotonin and neuropeptides are both released by the HSN command neuron to initiate Caenorhabditis elegans egg laying.

Neurons typically release both a small-molecule neurotransmitter and one or more neuropeptides, but how these two types of signal from the same neuron might act together remains largely obscure. For example, serotonergic neurons in mammalian brain express the neuropeptide Substance P, but it is unclear how this co-released neuropeptide might modulate serotonin signaling. We studied this issue in C. elegans, in which all serotonergic neurons express the neuropeptide NLP-3. The serotonergic Hermaphrodite Specific Neurons (HSNs) are command motor neurons within the egg-laying circuit which have been shown to release serotonin to initiate egg-laying behavior. We found that egg-laying defects in animals lacking serotonin were far milder than in animals lacking HSNs, suggesting that HSNs must release other signal(s) in addition to serotonin to stimulate egg laying. While null mutants for nlp-3 had only mild egg-laying defects, animals lacking both serotonin and NLP-3 had severe defects, similar to those of animals lacking HSNs. Optogenetic activation of HSNs induced egg laying in wild-type animals, and in mutant animals lacking either serotonin or NLP-3, but failed to induce egg laying in animals lacking both. We recorded calcium activity in the egg-laying muscles of animals lacking either serotonin, NLP-3, or both. The single mutants, and to a greater extent the double mutant, showed muscle activity that was uncoordinated and unable to expel eggs. Specifically, the vm2 muscles cells, which are direct postsynaptic targets of the HSN, failed to contract simultaneously with other egg-laying muscle cells. Our results show that the HSN neurons use serotonin and the neuropeptide NLP-3 as partially redundant co-transmitters that together stimulate and coordinate activity of the target cells onto which they are released.

Activity of the C. elegans egg-laying behavior circuit is controlled by competing activation and feedback inhibition.

Like many behaviors, Caenorhabditis elegans egg laying alternates between inactive and active states. To understand how the underlying neural circuit turns the behavior on and off, we optically recorded circuit activity in behaving animals while manipulating circuit function using mutations, optogenetics, and drugs. In the active state, the circuit shows rhythmic activity phased with the body bends of locomotion. The serotonergic HSN command neurons initiate the active state, but accumulation of unlaid eggs also promotes the active state independent of the HSNs. The cholinergic VC motor neurons slow locomotion during egg-laying muscle contraction and egg release. The uv1 neuroendocrine cells mechanically sense passage of eggs through the vulva and release tyramine to inhibit egg laying, in part via the LGC-55 tyramine-gated Cl- channel on the HSNs. Our results identify discrete signals that entrain or detach the circuit from the locomotion central pattern generator to produce active and inactive states.

The Effects of Forced Exhalation and Inhalation, Grunting, and Valsalva Maneuver on Forehand Force in Collegiate Tennis Players.

To examine the effects of forced expiration (FE), forced inspiration (FI), grunting (GR), and valsalva maneuver (VM), on air volume, maximum force production, and muscle recruitment during a simulated forehand stroke in collegiate tennis players. Superficial electrodes were placed over the anterior deltoid, pectoralis major, rectus abdominis, lumbar and thoracic erector spinae, and external and internal obliques. Subjects stood in a simulated forehand stroke stance with their dominant hand positioned on a force plate. Subjects performed 3 repetitions of maximal 2- to 3-second isometric forehands with randomly assigned breathing conditions (FE, FI, VM, and GR) and 30-seconds rest between contractions. Air volumes were also collected during each trial. A repeated-measures multivariate analysis of variance compared normalized peak electromyographic activity across the 7 muscle groups. Separate repeated measures ANOVAs compared the effect of breathing conditions on peak force production and air volume. Statistical significance was set at p ≤ 0.05 for all analyses. Overall muscle activity differed significantly by breathing condition (p = 0.031) with greater anterior deltoid activity was seen in FE and GR vs. VM. Internal oblique activity was significantly greater in GR than FI or VM, and thoracic erector spinae activity was significantly greater during FE and VM than FI. Force production did not differ significantly among breathing conditions (p = 0.74); however, GR forces were greater than FI and VM forces (p ≤ 0.05). Forced expiration air volume was significantly greater (p < 0.001) than FI, GR, or VM. These findings suggest that either GR or a more quiescent FE can be used to enhance force production. Forced expiration is a potentially safer alternative to VM and more aurally pleasing than GR.

The effect of stress on learning in surgical skill acquisition.

BACKGROUND: An excessive level of stress and anxiety in medical education can have a negative impact on learning. In particular, the interaction between attending surgeons and trainees in the operating room could induce stress on trainees that is counterproductive, especially if the teaching style or feedback is unduly harsh or critical. AIM: To characterize the effects of stress resulting from attending-trainee interaction during surgical skill acquisition. METHODS: Forty medical students learned to perform the FLS pattern-cutting task for the first time in one of four scenarios. In the control condition, no mentor was present. In the three experimental conditions, participants were observed, encouraged, or criticized by an expert surgeon. RESULTS: Task performance, as well as physiological and subjective indicators of stress, were measured. Taking both speed and accuracy into account, participants who were criticized performed the worst on the task, and those who were encouraged performed best. Physiological and subjective measures indicated that the criticized participants experienced the highest level of stress and anxiety. CONCLUSION: Even though providing constructive criticism to trainees is inevitable during the course of teaching, an exceedingly critical and negative mentoring style by attending physicians could be detrimental to trainees' acquisition of surgical skills.

Enriched open field facilitates exercise and social interaction in 2 strains of guinea pigs (Cavia porcellus).

Current housing guidelines for laboratory rodents include recommendations for enrichment. Working with guinea pigs, we have developed an open-field enrichment paradigm that provides several aspects of this species' natural environment. These naturalistic aspects include access to increased space for exploration, access to western timothy (Phleum pratense L.) hay, and grouping as a herd to facilitate social interaction. To determine the immediate effect on behavior from access to the enriched environment, female guinea pigs from 2 strains, IAF Hairless and NIH Hartley, were observed in both standard home cages and an open-field enriched environment. Subjects were housed with cagemates in pairs for the home-cage observation and were grouped as a herd when in the open-field arena. Behaviors were videorecorded for 1 h and then scored. Salivary cortisol levels were measured both prior to and immediately after behavioral observations. Analyses revealed higher levels of activity and social interaction in the open-field arena compared with the home cage, with no significant change in salivary cortisol levels. These results suggest that exposure to the open-field environment provide increased opportunities for exercise and social enrichment. Although additional studies are needed to determine long-term effects on experimental outcomes, the open-field configuration holds promise as a laboratory enrichment paradigm for guinea pigs.

Altered gene expression and spine density in nucleus accumbens of adolescent and adult male mice exposed to emotional and physical stress.

Stressful early life experiences are implicated in lifelong health. However, little is known about the consequences of emotional stress (ES) or physical stress (PS) on neurobiology. Therefore, the following set of experiments was designed to assess changes in transcription and translation of key proteins within the nucleus accumbens (NAc). Male adolescent (postnatal day 35) or adult (8-week-old) mice were exposed to ES or PS using a witness social defeat paradigm. Then, 24 h after the last stress session, we measured levels of specific mRNAs and proteins within the NAc. Spine density was also assessed in separate groups of mice. Exposure to ES or PS disrupted extracellular signal-related kinase 2 (ERK2), reduced transcription of ΔFosB and had no effect on cAMP response element-binding protein (CREB) mRNA. Western blots revealed that exposure to ES or PS decreased ERK2 phosphorylation in adolescents, whereas the same stress regimen increased ERK2 phosphorylation in adults. Exposure to ES or PS had no effect on ΔFosB or CREB phosphorylation. ES and PS increased spine density in the NAc of adolescent exposed mice, but only exposure to PS increased spine density in adults. Together, these findings demonstrate that exposure to ES or PS is a potent stressor in adolescent and adult mice and can disturb the integrity of the NAc by altering transcription and translation of important signaling molecules in an age-dependent manner. Furthermore, exposure to ES and PS induces substantial synaptic plasticity of the NAc.