Use of a Calving Blind That Imitates a Natural Environment.

Many indoor-housed cows isolate at calving when given the opportunity, and calving behaviors vary by blind and pen design. The objectives of this study were to determine if cows preferred calving in a visibly separated (blind) or an open area of a group maternity pen, and if there was a preference for the degree of seclusion provided by the blind (50% vs. 100% coverage). Two calving blinds were provided in a group calving pen, and the amount of visibility through the blinds was created using firehoses secured from the top of a metal frame that lined the entire front of the blind (100%) or with every other hose rolled up (50%). Holstein cows and heifers (n = 79) were enrolled into a dynamic group calving pen 21 ± 3 d before calving. Calving location, the difference in blind use prior to calving compared to a baseline period, and social behaviors were recorded using video observation. There was no difference in the number of cows that calved in or outside of a blind (28 vs. 37 calvings, respectively). Cows were more likely to calve in a blind during the day than at night and as the number of cows in the pen increased. For cows who calved in a blind, there was no preference for calving in the 50% or 100% blind (10 vs. 18, respectively). Providing a varied environment for intensively managed cattle can improve their welfare by allowing cows the opportunity to perform natural behaviors and choice over their environment.

Quantitative somatosensory testing of the abdomen: establishing initial reference values across developmental age and biological sex.

ABSTRACT: Abdominal pain is a common symptom of several debilitating conditions (eg, inflammatory bowel disease, irritable bowel syndrome, and endometriosis) and affects individuals throughout their lifespan. Quantitative sensory testing (QST) reference values exist for many body sites but not the abdomen. Using a QST battery adapted from the German Research Network on Neuropathic Pain, we collected QST data on the upper and lower abdomen in 181 pain-free participants, ages 12 to 50 years, to establish reference values by age and biological sex. The normative values are presented as medians for each QST measure by sex (male, n = 63; female, n = 118) and across 3 age categories (adolescents: 12-19 years, n = 48; young adults: 20-30 years, n = 87; and adults: 31-50 years, n = 46). Evaluating the sensory functioning of the abdomen and characterizing ranges of QST measures is an essential first step in understanding and monitoring the clinical course of sensory abnormalities in patients with underlying diseases affecting the abdomen and pelvis. The impact of age and development on sensory functioning is necessary, given age-related changes in pain perception and modulation.

Evaluation of prevalence and outcomes of serial tyrosine kinase inhibitor use in pediatric patients with advanced solid tumors.

PURPOSE: Multitargeted tyrosine kinase inhibitors (mTKIs) are increasingly utilized in the treatment of pediatric sarcomas and other solid tumors. It is unknown whether serial treatment with multiple TKIs provides a benefit and which patients are most likely to benefit from mTKI rechallenge. METHODS: We performed a retrospective cohort study of pediatric cancer patients who received serial mTKI therapy off-study between 2007 and 2020 as either monotherapy or combination therapy. We report patient characteristics, clinical outcomes, dosing patterns, and treatment-associated toxicity. RESULTS: The study cohort included 25 patients. The overall prevalence of serial mTKI therapy among all patients treated for sarcoma at our institution was 3.7%, and the response rate to second mTKI was 9%. Median 6-month progression-free survival (PFS) and overall survival (OS) from start of second mTKI were 42.1% (95% CI: 20.4%-62.5%) and 79.1% (95% CI: 57.0%-90.8%), respectively. Patients who had received 4 months or more (n = 11) of therapy with first mTKI had significantly longer PFS versus those who received less than 4 months (n = 11; p = .001). Thirty-three percent of patients discontinued second mTKI due to toxicity. Six (40%) of 15 patients who discontinued the first mTKI due to progression had either a partial response or stable disease on the second mTKI. CONCLUSIONS: We observed a low response rate to mTKI rechallenge. However, we identified patients who had been treated with first mTKI for  ≥4 months as more likely to have prolonged stable disease with second mTKI. Several patients had a response or stable disease on the second mTKI despite having progressed on the first mTKI. Though toxicity was common, only a minority of patients discontinued the second mTKI due to toxicity.

Broken tails in Holstein dairy cattle: A cross-sectional study.

Dairy cows are regularly handled when moved to the milking parlor and during other routine procedures. Low-stress handling methods are important in avoiding negative welfare states for dairy cattle. Tail twisting is used by some handlers to prompt cattle movement. However, when used inappropriately with excessive force, tail twisting can lead to a broken tail. The aim of this cross-sectional study was to determine cow-level factors that may be associated with the prevalence of broken tails in dairy cattle. A subset of 229 Holstein dairy cows (68 primiparous and 161 multiparous) at a single dairy were assessed for broken tails from the larger herd (N = 1,356). Tails were visually assessed for the presence of fractures by a single trained observer. A tail was classified as unfractured if it laid straight when at rest and as fractured if there were deviations in the tail when at rest. Poisson regression models were used to identify associations between cow-level characteristics and broken tails and compute adjusted prevalence ratios (PR). The prevalence of broken tails was 45.8% (105/229) at the time of assessment. Multiparous cows had a greater prevalence of broken tails than primiparous cows [PR = 1.70; 95% confidence interval (CI): 1.11-2.59]. The prevalence of broken tails was also greater for cows treated for mastitis ≥2 times than cows treated once for mastitis (PR = 1.84; 95% CI: 1.08-3.13) and cows never treated for mastitis (PR = 1.36; 95% CI: 1.02-1.82). Results from this study indicated that the longer a cow was present on the farm and the more times she was treated for mastitis, the more likely she was to experience a broken tail. These findings suggest that the relationship between dairy cow handling, health, and welfare is a multifactorial issue.

Immunotherapy approaches for the treatment of diffuse midline gliomas.

Diffuse midline gliomas (DMG) are a highly aggressive and universally fatal subgroup of pediatric tumors responsible for the majority of childhood brain tumor deaths. Median overall survival is less than 12 months with a 90% mortality rate at 2 years from diagnosis. Research into the underlying tumor biology and numerous clinical trials have done little to change the invariably poor prognosis. Continued development of novel, efficacious therapeutic options for DMGs remains a critically important area of active investigation. Given that DMGs are not amenable to surgical resection, have only limited response to radiation, and are refractory to traditional chemotherapy, immunotherapy has emerged as a promising alternative treatment modality. This review summarizes the various immunotherapy-based treatments for DMG as well as their specific limitations. We explore the use of cell-based therapies, oncolytic virotherapy or immunovirotherapy, immune checkpoint inhibition, and immunomodulatory vaccination strategies, and highlight the recent clinical success of anti-GD2 CAR-T therapy in diffuse intrinsic pontine glioma (DIPG) patients. Finally, we address the challenges faced in translating preclinical and early phase clinical trial data into effective standardized treatment for DMG patients.

Standard clinical approaches and emerging modalities for glioblastoma imaging.

Glioblastoma (GBM) is the most common primary adult intracranial malignancy and carries a dismal prognosis despite an aggressive multimodal treatment regimen that consists of surgical resection, radiation, and adjuvant chemotherapy. Radiographic evaluation, largely informed by magnetic resonance imaging (MRI), is a critical component of initial diagnosis, surgical planning, and post-treatment monitoring. However, conventional MRI does not provide information regarding tumor microvasculature, necrosis, or neoangiogenesis. In addition, traditional MRI imaging can be further confounded by treatment-related effects such as pseudoprogression, radiation necrosis, and/or pseudoresponse(s) that preclude clinicians from making fully informed decisions when structuring a therapeutic approach. A myriad of novel imaging modalities have been developed to address these deficits. Herein, we provide a clinically oriented review of standard techniques for imaging GBM and highlight emerging technologies utilized in disease characterization and therapeutic development.

Targeting oncometabolism to maximize immunotherapy in malignant brain tumors.

Brain tumors result in significant morbidity and mortality in both children and adults. Recent data indicate that immunotherapies may offer a survival benefit after standard of care has failed for malignant brain tumors. Modest results from several late phase clinical trials, however, underscore the need for more refined, comprehensive strategies that incorporate new mechanistic and pharmacologic knowledge. Recently, oncometabolism has emerged as an adjunct modality for combinatorial treatment approaches necessitated by the aggressive, refractory nature of high-grade glioma and other progressive malignant brain tumors. Manipulation of metabolic processes in cancer and immune cells that comprise the tumor microenvironment through controlled targeting of oncogenic pathways may be utilized to maximize the efficacy of immunotherapy and improve patient outcomes. Herein, we summarize preclinical and early phase clinical trial research of oncometabolism-based therapeutics that may augment immunotherapy by exploiting the biochemical and genetic underpinnings of brain tumors. We also examine metabolic pathways related to immune cells that target tumor cells, termed "tumor immunometabolism". Specifically, we focus on glycolysis and altered glucose metabolism, including glucose transporters, hexokinase, pyruvate dehydrogenase, and lactate dehydrogenase, glutamine, and we discuss targeting arginase, adenosine, and indoleamine 2,3-dioxygenase, and toll-like receptors. Lastly, we summarize future directions targeting metabolism in combination with emerging therapies such as oncolytic virotherapy, vaccines, and chimeric antigen receptor T cells.

Therapeutic cancer vaccines for pediatric malignancies: advances, challenges, and emerging technologies.

Though outcomes for pediatric cancer patients have significantly improved over the past several decades, too many children still experience poor outcomes and survivors suffer lifelong, debilitating late effects after conventional chemotherapy, radiation, and surgical treatment. Consequently, there has been a renewed focus on developing novel targeted therapies to improve survival outcomes. Cancer vaccines are a promising type of immunotherapy that leverage the immune system to mediate targeted, tumor-specific killing through recognition of tumor antigens, thereby minimizing off-target toxicity. As such, cancer vaccines are orthogonal to conventional cancer treatments and can therefore be used alone or in combination with other therapeutic modalities to maximize efficacy. To date, cancer vaccination has remained largely understudied in the pediatric population. In this review, we discuss the different types of tumor antigens and vaccine technologies (dendritic cells, peptides, nucleic acids, and viral vectors) evaluated in clinical trials, with a focus on those used in children. We conclude with perspectives on how advances in combination therapies, tumor antigen (eg, neoantigen) selection, and vaccine platform optimization can be translated into clinical practice to improve outcomes for children with cancer.

Quantification of cooling effects on basic tissue measurements and exposed cross-sectional brain area of cadaver heads from market pigs.

The objective of this project was to determine the impact of cooling on the soft tissue thickness, cranial thickness, and cross-sectional brain area of cadaver heads from market pigs. Documenting the effect of cooling on tissue dimensions of swine heads is valuable and important for future investigations of physical stunning and euthanasia methods that use cadaver heads. Scalded and dehaired cadaver heads with intact jowls were sourced from market pigs stunned with CO2 gas. After transport to the data collection location, a penetrating captive bolt (PCB) shot (Jarvis Model PAS-Type P 0.25R Caliber Captive Bolt Pistol with Medium Rod Assembly and Blue Powder Cartridges) was applied in the frontal position. Following PCB application, each head (n = 36) underwent an UNCHILLED treatment followed by CHILLED treatment. The UNCHILLED treatment involved images collected immediately after splitting each head along the bolt path, and the CHILLED treatment involved images of the same heads after storage in a walk-in cooler for 24 h at 2 to 4°C. All measurements for each treatment were collected from images of the heads on the plane of the bolt path immediately prior to and immediately after the refrigeration treatment. Measurements were performed by two observers. Across all measurements, mean interobserver coefficient of variation was 11.3 ± 0.6%. The soft tissue caudal to the bolt path was different (P = 0.0120) between treatments (CHILLED: 6.4 ± 0.2 mm; UNCHILLED: 7.2 ± 0.2 mm). The soft tissue thickness rostral to the bolt path was different (P = 0.0378) between treatments (CHILLED: 5.5 ± 0.2 mm; UNCHILLED: 6.1 ± 0.2 mm). Cranial thickness caudal to the bolt path was not different (P = 0.8659; CHILLED: 18.1 ± 0.6 mm; UNCHILLED: 18.3 ± 0.6 mm), nor was there a significant difference (P = 0.2593) in cranial thickness rostral to the bolt path between treatments (CHILLED: 16.2 ± 0.6 mm; UNCHILLED: 15.2 ± 0.6 mm). Cross-sectional brain area did not differ (P = 0.0737; CHILLED: 3633.4 ± 44.1 mm; UNCHILLED: 3519.9 ± 44.1 mm). A correction factor of 1.12 was determined from this study for cases where estimation of UNCHILLED soft tissue thickness from CHILLED soft tissue thickness is necessary.

Author Correction: Striatal neurons directly converted from Huntington's disease patient fibroblasts recapitulate age-associated disease phenotypes.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Trends in conditional survival and predictors of late death in neuroblastoma.

PURPOSE: Significant advances in the treatment of neuroblastoma have been made in the past several decades. There are scant data examining how these improvements have changed over time and differentially affected conditional survival among high-risk and non-high-risk patient groups. METHODS: We conducted a retrospective cohort study using the Surveillance, Epidemiology, and End Results database. We analyzed clinical characteristics and survival outcomes for 4717 neuroblastoma patients. Kaplan-Meier methods were used to estimate overall survival (OS) and conditional overall survival (COS) with estimates compared between groups using log-rank tests. RESULTS: Five-year OS was 41.46% (95% CI 38.77-44.13) for the high-risk group and 91.13% (95% CI 89.49-92.53) for the non-high-risk group. Both groups saw significant improvements in OS by decade (P < .001). Five-year COS among 1-year survivors was 52.69% (CI 49.54-55.73) for the high-risk group and 96.75% (95% CI 95.57-97.62) for the non-high-risk group. One-year survivors in the high-risk group showed a statistically significant improvement in COS over time. No difference in COS was observed among 5-year high-risk survivors. In the high-risk and non-high-risk groups, 82% and 32% of late deaths were attributable to cancer, respectively. Statistically significant adverse prognostic factors for late death were age ≥ 1 year at diagnosis, metastatic disease, and nonthoracic primary site (P = .001). CONCLUSIONS: Improvements in COS over time have largely benefited high-risk patients, though they are still at higher risk for late death due to cancer when compared to non-high-risk patients. Age, stage, and primary site, but not treatment decade, influence outcomes among 5-year survivors.

Corpus Callosotomy for Refractory Epilepsy in Aicardi Syndrome: Case Report and Focused Review of the Literature.

BACKGROUND: Aicardi syndrome is a severe neurodevelopmental disorder that occurs primarily in females and is characterized by seizures, agenesis of the corpus callosum, and chorioretinal lacunae, which occur together in the majority of affected individuals. Seizures begin in infancy and tend to progress in intensity and are often refractory to standard multimodal medication treatments. CASE DESCRIPTION: We present here a unique case of a 12-year-old girl with partial agenesis of the corpus callosum who underwent a corpus callosotomy for treatment of medically refractory epilepsy. In so doing, we also review the literature with regard to the neurosurgical management of these unique patients. CONCLUSIONS: For the subset of children who present with partial, rather than complete, agenesis of the corpus callosum, corpus callosotomy should be considered as a treatment option to reduce seizure burden.

Suppression of proteolipid protein rescues Pelizaeus-Merzbacher disease.

Mutations in PLP1, the gene that encodes proteolipid protein (PLP), result in failure of myelination and neurological dysfunction in the X-chromosome-linked leukodystrophy Pelizaeus-Merzbacher disease (PMD)1,2. Most PLP1 mutations, including point mutations and supernumerary copy variants, lead to severe and fatal disease. Patients who lack PLP1 expression, and Plp1-null mice, can display comparatively mild phenotypes, suggesting that PLP1 suppression might provide a general therapeutic strategy for PMD1,3-5. Here we show, using CRISPR-Cas9 to suppress Plp1 expression in the jimpy (Plp1jp) point-mutation mouse model of severe PMD, increased myelination and restored nerve conduction velocity, motor function and lifespan of the mice to wild-type levels. To evaluate the translational potential of this strategy, we identified antisense oligonucleotides that stably decrease the levels of Plp1 mRNA and PLP protein throughout the neuraxis in vivo. Administration of a single dose of Plp1-targeting antisense oligonucleotides in postnatal jimpy mice fully restored oligodendrocyte numbers, increased myelination, improved motor performance, normalized respiratory function and extended lifespan up to an eight-month end point. These results suggest that PLP1 suppression could be developed as a treatment for PMD in humans. More broadly, we demonstrate that oligonucleotide-based therapeutic agents can be delivered to oligodendrocytes in vivo to modulate neurological function and lifespan, establishing a new pharmaceutical modality for myelin disorders.

Relationship of captive bolt stunning location with basic tissue measurements and exposed cross-sectional brain area in cadaver heads from market pigs.

The objective of this study was to contrast the soft tissue thickness, cranial thickness, total tissue thickness, cross-sectional brain area, and bolt-brain contact from the common frontal application of captive bolt euthanasia with the alternative location behind the ear in cadaver swine heads. Twenty-three cadaver heads from pigs that were approximately 136 kg and 6 mo of age were collected from a regional slaughter establishment following CO2 stunning and assigned to either the FRONTAL (n = 11) or the CAUDAL TO PINNA (n = 12) application of the captive bolt. The soft tissue thickness was different (P < 0.0001) between the 2 applications (FRONTAL: 8.3 ± 3.4 mm; CAUDAL TO PINNA: 56.5 ± 3.4 mm). The cranial thickness was different (P < 0.0001) between the applications (FRONTAL: 23.4 ± 2.9 mm; CAUDAL TO PINNA: 26.5 ± 2.9 mm). There was also a difference (P < 0.0001) in the total tissue thickness between the 2 applications (FRONTAL: 31.7 ± 3.8 mm; CAUDAL TO PINNA: 73.4 ± 3.8 mm). Cross-sectional area was calculated from images collected immediately after the heads were cut along the plane of bolt travel by bandsaw and was different (P = 0.0028) between the 2 applications (FRONTAL: 25.2 ± 1.3 cm2; CAUDAL TO PINNA: 18.9 ± 1.3 cm2). Bolt-brain contact was also assessed from the images, and a difference (P = 0.0360) between the 2 applications (FRONTAL: 100 ± 10.5%; CAUDAL TO PINNA: 66.7 ± 10.5%) was identified. The results of this study suggest that the FRONTAL application may provide a bolt path with less tissue to travel through when compared with the CAUDAL TO PINNA application for pigs of the approximate age and weight of those in this study. Ultimately, the FRONTAL location may present less risk for the captive bolt euthanasia of swine at market weight at this time. Additional refinement of the CAUDAL TO PINNA procedure and modification to the captive bolt device to penetrate to a suitable depth to ensure brain damage is recommended.

Rapid functional genetics of the oligodendrocyte lineage using pluripotent stem cells.

Oligodendrocyte dysfunction underlies many neurological disorders, but rapid assessment of mutation-specific effects in these cells has been impractical. To enable functional genetics in oligodendrocytes, here we report a highly efficient method for generating oligodendrocytes and their progenitors from mouse embryonic and induced pluripotent stem cells, independent of mouse strain or mutational status. We demonstrate that this approach, when combined with genome engineering, provides a powerful platform for the expeditious study of genotype-phenotype relationships in oligodendrocytes.

Striatal neurons directly converted from Huntington's disease patient fibroblasts recapitulate age-associated disease phenotypes.

In Huntington's disease (HD), expansion of CAG codons in the huntingtin gene (HTT) leads to the aberrant formation of protein aggregates and the differential degeneration of striatal medium spiny neurons (MSNs). Modeling HD using patient-specific MSNs has been challenging, as neurons differentiated from induced pluripotent stem cells are free of aggregates and lack an overt cell death phenotype. Here we generated MSNs from HD patient fibroblasts through microRNA-based direct neuronal conversion, bypassing the induction of pluripotency and retaining age signatures of the original fibroblasts. We found that patient MSNs consistently exhibited mutant HTT (mHTT) aggregates, mHTT-dependent DNA damage, mitochondrial dysfunction and spontaneous degeneration in culture over time. We further provide evidence that erasure of age stored in starting fibroblasts or neuronal conversion of presymptomatic HD patient fibroblasts results in differential manifestation of cellular phenotypes associated with HD, highlighting the importance of age in modeling late-onset neurological disorders.