Characterisation of LPS+ bacterial extracellular vesicles along the gut-hepatic portal vein-liver axis.

Gut microbiome dysbiosis is a major contributing factor to several pathological conditions. However, the mechanistic understanding of the communication between gut microbiota and extra-intestinal organs remains largely elusive. Extracellular vesicles (EVs), secreted by almost every form of life, including bacteria, could play a critical role in this inter-kingdom crosstalk and are the focus of present study. Here, we present a novel approach for isolating lipopolysaccharide (LPS)+ bacterial extracellular vesicles (bEVLPS) from complex biological samples, including faeces, plasma and the liver from lean and diet-induced obese (DIO) mice. bEVLPS were extensively characterised using nanoparticle tracking analyses, immunogold labelling coupled with transmission electron microscopy, flow cytometry, super-resolution microscopy and 16S sequencing. In liver tissues, the protein expressions of TLR4 and a few macrophage-specific biomarkers were assessed by immunohistochemistry, and the gene expressions of inflammation-related cytokines and their receptors (n = 89 genes) were measured using a PCR array. Faecal samples from DIO mice revealed a remarkably lower concentration of total EVs but a significantly higher percentage of LPS+ EVs. Interestingly, DIO faecal bEVLPS showed a higher abundance of Proteobacteria by 16S sequencing. Importantly, in DIO mice, a higher number of total EVs and bEVLPS consistently entered the hepatic portal vein and subsequently reached the liver, associated with increased expression of TLR4, macrophage markers (F4/80, CD86 and CD206), cytokines and receptors (Il1rn, Ccr1, Cxcl10, Il2rg and Ccr2). Furthermore, a portion of bEVLPS escaped liver and entered the peripheral circulation. In conclusion, bEV could be the key mediator orchestrating various well-established biological effects induced by gut bacteria on distant organs.

Experiences with pregnancy and child-rearing, advice for future mothers, and suggestions for improving the system: a qualitative study of women in neurosurgery.

OBJECTIVE: Neurosurgery is among the most demanding and time-consuming occupations, and with diversity and inclusion initiatives only recently increasing the number of women in the field, efforts still need to be made to help neurosurgery become more accommodating for pregnancy and child-rearing. Thus, the present study sought to be the first to investigate this issue through in-depth qualitative interviews of women in neurosurgery. METHODS: A total of 33 female neurosurgeons participated in semistructured Zoom interviews. Cocoding and thematic analysis were conducted with interview transcripts to determine themes and corresponding subthemes with regard to these women's experiences with pregnancy and child-rearing, advice for future mothers in neurosurgery, and suggestions for improving the field of neurosurgery for those desiring children. RESULTS: Among the 33 participants, 22 (66.7%) had given birth to or adopted at least one child, had at least one stepchild, or were pregnant at the time of the interview. Three themes emerged regarding these 22 women's experiences with pregnancy and child-rearing: 1) challenges with the physiological changes of pregnancy, 2) feelings of guilt and anxiety, and 3) reliance on loved ones for childcare. Three themes emerged among these 22 women's advice for future mothers in neurosurgery: 1) set realistic expectations, 2) take control of your schedule, and 3) realize that there is no "right" time to start a family. Finally, two themes emerged among all 33 participants' suggestions for making neurosurgery more feasible for pregnancy and child-rearing: 1) revamping of on-site resources, and 2) improved guidance on family planning, childbearing, and maternity leave. The most prominent subtheme in the authors' study was a call for improved on-site daycare under the "revamping of on-site resources" theme, with a particular emphasis on 24/7 operation. CONCLUSIONS: The authors' data have illustrated the themes of the experiences and thoughts of women in a field where pregnancy and child-rearing are arguably the most challenging of any occupation. Resources such as improved on-site daycare and organized, program-specific information sets for future mothers appear to comprise a consensus of suggested solutions by the women directly experiencing these challenges. The authors' results may be useful in guiding system-wide changes that may improve the field of neurosurgery for current and future mothers.

Effects of protein concentration and beta-adrenergic agonists on ruminal bacterial communities in finishing beef heifers.

To improve animal performance and modify growth by increasing lean tissue accretion, beef cattle production has relied on use of growth promoting technologies such as beta-adrenergic agonists. These synthetic catecholamines, combined with the variable inclusion of rumen degradable (RDP) and undegradable protein (RUP), improve feed efficiency and rate of gain in finishing beef cattle. However, research regarding the impact of beta-adrenergic agonists, protein level, and source on the ruminal microbiome is limited. The objective of this study was to determine the effect of different protein concentrations and beta-adrenergic agonist (ractopamine hydrochloride; RAC) on ruminal bacterial communities in finishing beef heifers. Heifers (n = 140) were ranked according to body weight and assigned to pens in a generalized complete block design with a 3 × 2 factorial arrangement of treatments of 6 different treatment combinations, containing 3 protein treatments (Control: 13.9% CP, 8.9% RDP, and 5.0% RUP; High RDP: 20.9% CP, 14.4% RDP, 6.5% RUP; or High RUP: 20.9% CP, 9.7% RDP, 11.2% RUP) and 2 RAC treatments (0 and 400 mg/day). Rumen samples were collected via orogastric tubing 7 days before harvest. DNA from rumen samples were sequenced to identify bacteria based on the V1-V3 hypervariable regions of the 16S rRNA gene. Reads from treatments were analyzed using the packages 'phyloseq' and 'dada2' within the R environment. Beta diversity was analyzed based on Bray-Curtis distances and was significantly different among protein and RAC treatments (P < 0.05). Alpha diversity metrics, such as Chao1 and Shannon diversity indices, were not significantly different (P > 0.05). Bacterial differences among treatments after analyses using PROC MIXED in SAS 9 were identified for the main effects of protein concentration (P < 0.05), rather than their interaction. These results suggest possible effects on microbial communities with different concentrations of protein but limited impact with RAC. However, both may potentially act synergistically to improve performance in finishing beef cattle.

Association Between Social Determinants of Health, Distance from Treatment Center, and Treatment Type with Outcomes in Human Papillomavirus Associated Oropharyngeal cancer.

OBJECTIVES: Social determinants of health (SDOH) can influence access to cancer care, clinical trials, and oncologic outcomes. We investigated the association between SDOH, distance from treatment center, and treatment type with outcomes in human papillomavirus associated oropharyngeal squamous cell carcinoma [HPV(+)OPSCC] patients treated at a tertiary care center. STUDY DESIGN: Retrospective review. METHODS: HPV(+)OPSCC patients treated surgically from 2006 to 2021 were selected from our departmental Oropharyngeal Cancer RedCap database. Demographic data, treatment, and oncologic outcomes were extracted. Distance was calculated in miles between the centroid of each patient zip code and our hospital zip code (zipdistance). RESULTS: 874 patients (89 % male; mean age: 58 years) were identified. Most patients (96 %) reported Non-Hispanic White as their primary race. 204 patients (23 %) had a high-school degree or less, 217 patients (25 %) reported some college education or a 2-year degree, 153 patients (18 %) completed a four-year college degree, and 155 patients (18 %) had post-graduate degrees. Relative to those with a high-school degree, patients with higher levels of education were more likely to live further away from our institution (p < 0.0001). Patients who received adjuvant radiation therapy elsewhere lived, on average, 104 miles further away than patients receiving radiation at our institution (Estimate 104.3, 95 % CI 14.2-194.4, p-value = 0.02). In univariable Cox PH models, oncologic outcomes did not significantly differ by zipdistance. CONCLUSIONS: Education level-and access to resources-varied proportionally to a patient's distance from our center. Patients travelling further distances for surgical management of OPSCC were more likely to pursue adjuvant radiation therapy at an outside institution. Distance traveled was not associated with oncologic outcomes. Breaking down barriers to currently excluded populations may improve access to clinical trials and improve oncologic outcomes for diverse patient populations.

Poverty and the wellbeing of children with sickle cell disease: The mediating role of parenting stress.

BACKGROUND: Living in poverty in the midst of caring for a child with sickle cell disease (SCD) presents with challenges that may be overwhelming for caregivers. Prior research has shown a relationship between poverty and poor outcomes in children; however, no study has investigated the mediating role of parenting stress on health-related quality of life (HRQOL) for such children. PROCEDURE: A total of 150 patients, ages 8-17 years and their parents, were enrolled and completed measures of parenting stress (caregivers) and HRQOL (patients). Caregivers also completed a demographic information questionnaire, which was used to determine poverty status. The Sobel test was used to analyze the association between poverty and HRQOL by way of parenting stress. RESULTS: The series of linear regression results show poverty was a significant predictor for decreased HRQOL (B = -7.01, p = .05) among pediatric patients with SCD. Moreover, the regression coefficient measuring the indirect effect of the model measuring the mediating role of parenting stress was statistically significant (B = -1.05, p = .05). CONCLUSION: The current study found an association between poverty status and HRQOL scores by way of parenting stress. Findings highlight the need for appropriate resource allocation to caregivers in addition to pediatric patients living with SCD.

Family conflict over illness beliefs and care strategies: implications for burden in family caregivers.

OBJECTIVES: Family conflict is an understudied aspect of the caregiver experience. Drawing from the stress process model, the present study examined the prevalence and correlates of conflict over illness beliefs (e.g. conflict over the care recipient's illness and need for facility placement) and family care strategies (e.g. lack of involvement and support from family members). METHODS: Adult child and spouse caregivers (N = 579) of persons with physical or cognitive impairment from clinic and internet samples completed the Caregiver Reaction Scale (CRS) to assess each topic of conflict and burden. RESULTS: Most caregivers reported conflict with family members over care strategies (63%) or illness beliefs (55%). Clinic caregivers reported greater illness beliefs conflict than online caregivers. Adult children reported greater illness beliefs and family care strategies than spouses. Male caregivers were more likely than female caregivers to report care strategies conflict. Caregiver overload was significantly associated with family care strategies conflict. Both conflicts were positively associated with caregiver burden. CONCLUSIONS: Findings further support that family conflict is a widespread concern associated with burden among online and help-seeking caregivers. Caregiving relationship type and gender may increase likelihood of each conflict. Findings inform recommendations for clinical intervention and assessment of family caregivers.

A Novel Metastatic Estrogen Receptor-Expressing Breast Cancer Model with Antiestrogen Responsiveness.

Most women diagnosed with breast cancer (BC) have estrogen receptor alpha-positive (ER+) disease. The current mouse models of ER+ BC often rely on exogenous estrogen to encourage metastasis, which modifies the immune system and the function of some tissues like bone. Other studies use genetically modified or immunocompromised mouse strains, which do not accurately replicate the clinical disease. To create a model of antiestrogen responsive BC with spontaneous metastasis, we developed a mouse model of 4T1.2 triple-negative (TN) breast cancer with virally transduced ER expression that metastasizes spontaneously without exogenous estrogen stimulation and is responsive to antiestrogen drugs. Our mouse model exhibited upregulated ER-responsive genes and multi-organ metastasis without exogenous estrogen administration. Additionally, we developed a second TN BC cell line, E0771/bone, to express ER, and while it expressed ER-responsive genes, it lacked spontaneous metastasis to clinically important tissues. Following antiestrogen treatment (tamoxifen, ICI 182,780, or vehicle control), 4T1.2- and E0771/bone-derived tumor volumes and weights were significantly decreased, exemplifying antiestrogen responsivity in both cell lines. This 4T1.2 tumor model, which expresses the estrogen receptor, metastasizes spontaneously, and responds to antiestrogen treatment, will allow for further investigation into the biology and potential treatment of metastasis.

Viral Disinfection of Porous Fomites Utilizing a Bacteriophage Model and Chlorine Dioxide Gas.

The pursuit of disinfecting porous materials or fomites to inactivate viral agents has special challenges. To address these challenges, a highly portable chlorine dioxide (ClO2) gas generation system was used to ascertain the ability of a gaseous preparation to inactivate a viral agent, the MS2 bacteriophage, when associated with potentially porous fomites of cloth, paper towel, and wood. The MS2 bacteriophage is increasingly used as a model to identify means of inactivating infectious viral agents of significance to humans. Studies showed that MS2 bacteriophage can be applied to and subsequently recovered from potential porous fomites such as cloth, paper towel, and wood. Paired with viral plaque assays, this provided a means for assessing the ability of gaseous ClO2 to inactivate bacteriophage associated with the porous materials. Notable results include 100% inactivation of 6 log bacteriophage after overnight exposure to 20 parts per million(ppm) ClO2. Reducing exposure time to 90 minutes and gas ppm to lower concentrations proved to remain effective in bacteriophage elimination in association with porous materials. Stepwise reduction in gas concentration from 76 ppm to 5 ppm consistently resulted in greater than 99.99% to 100% reduction of recoverable bacteriophage. This model suggests the potential of ClO2 gas deployment systems for use in the inactivation of viral agents associated with porous potential fomites. The ClO2 gas could prove especially helpful in disinfecting enclosed areas containing viral contaminated surfaces, rather than manually spraying and wiping them.

A Longitudinal Study on the Dynamics of Salmonella enterica Prevalence and Serovar Composition in Beef Cattle Feces and Lymph Nodes and Potential Contributing Sources from the Feedlot Environment.

Salmonella can persist in the feedlot pen environment, acting as a source of transmission among beef cattle. Concurrently, cattle that are colonized with Salmonella can perpetuate contamination of the pen environment through fecal shedding. To study these cyclical dynamics, pen environment and bovine samples were collected for a 7-month longitudinal comparison of Salmonella prevalence, serovar, and antimicrobial resistance profiles. These samples included composite environment, water, and feed from the feedlot pens (n = 30) and cattle (n = 282) feces and subiliac lymph nodes. Salmonella prevalence across all sample types was 57.7%, with the highest prevalence in the pen environment (76.0%) and feces (70.9%). Salmonella was identified in 42.3% of the subiliac lymph nodes. Based on a multilevel mixed-effects logistic regression model, Salmonella prevalence varied significantly (P < 0.05) by collection month for most sample types. Eight Salmonella serovars were identified, and most isolates were pansusceptible, except for a point mutation in the parC gene, associated with fluoroquinolone resistance. There was a proportional difference in serovars Montevideo, Anatum, and Lubbock comparing the environment (37.2, 15.9, and 11.0%, respectively), fecal (27.5, 22.2, and 14.6%, respectively), and lymph node (15.6, 30.2, and 17.7%, respectively) samples. This suggests that the ability of Salmonella to migrate from the pen environment to the cattle host-or vice versa-is serovar specific. The presence of certain serovars also varied by season. Our results provide evidence that Salmonella serovar dynamics differ when comparing environment and host; therefore, developing serovar-specific preharvest environmental Salmonella mitigation strategies should be considered. IMPORTANCE Salmonella contamination of beef products, specifically from the incorporation of bovine lymph nodes into ground beef, remains a food safety concern. Current postharvest Salmonella mitigation techniques do not address Salmonella bacteria that are harbored in the lymph nodes, nor is it well understood how Salmonella invades the lymph nodes. Alternatively, preharvest mitigation techniques that can be applied to the feedlot environment, such as moisture applications, probiotics, or bacteriophage, may reduce Salmonella before dissemination into cattle lymph nodes. However, previous research conducted in cattle feedlots includes study designs that are cross-sectional, are limited to point-in-time sampling, or are limited to sampling of the cattle host, making it difficult to assess the Salmonella interactions between environment and hosts. This longitudinal analysis of the cattle feedlot explores the Salmonella dynamics between the feedlot environment and beef cattle over time to determine the applicability of preharvest environmental treatments.

Impacts of including Sweet Bran and wet distillers grains with solubles alone or in combination in finishing cattle diets on physically effective fiber concentrations and rumen buffering characteristics of feedlot cattle.

This study evaluated the effects of Sweet Bran (SB) and wet distillers grains with solubles (WDGS) in the diet alone or in combination on physically effective neutral detergent fiber (peNDF), ruminal pH, and rumination behavior of finishing beef cattle. For this study, 455 steers (373 ± 15.5 kg) were allocated to 48 pens in a randomized complete block design. Treatments (n = 12 pens per treatment) were one of four steam-flaked corn-based diets containing no corn-milling products (CON), 20% WDGS (WDGS20), 20% SB (SB20), or 20% SB and 10% WDGS (COMBO). Within each pen, two steers were randomly selected to receive an indwelling ruminal pH bolus to quantify ruminal pH and a 3-axis accelerometer tag to measure rumination for the first 92 d of the study. Diet samples were collected weekly to determine particle size, neutral detergent fiber (NDF) concentration, and peNDF. Physically effective NDF was calculated using both the proportion of particles > 4.0 mm (peNDF4.0 ) and the proportion of particles > 8.0 mm (peNDF8.0). The percentage of particles > 4.0 mm was greatest (P < 0.01) for CON, intermediate for SB20, and least for WDGS20 and COMBO. Both NDF (P < 0.01) and peNDF4.0 (P < 0.01) were greatest for COMBO, intermediate for WDGS20 and SB20, and least for CON. The percentage of particles > 8.0 was greatest (P < 0.01) for CON, intermediate for WDGS20 and SB20, and least for COMBO, but peNDF8.0 did not differ (P = 0.40). A diet × day interaction (P < 0.01) was observed for daily rumination minutes per kg of DMI, NDF, peNDF4.0, and peNDF8.0. A diet × hour interaction (P < 0.01) was observed where CON cattle spent less time ruminating at 0800 and 1000 h in a 24 h period. Daily ruminal pH was greatest (P < 0.01) for COMBO, intermediate for SB20 and WDGS20, and least for CON. A diet × hour interaction (P < 0.01) was also observed for circadian ruminal pH, where pH was least for CON from 0800 to 1800 h. Relationships between peNDF, rumination behavior, and ruminal pH observed in this study suggest that SB and WDGS similarly enhance rumen buffering capacity when steam-flaked corn is replaced in the diet.

Management of nonoperative cervical spine fractures: An institutional analysis of follow-up duration and image acquisition.

Background: How long do nonoperative cervical fractures have to be followed and with how many imaging studies? Methods: We reviewed 69 adult patients with 122 (31 occiptocervical and 91 sub-axial) cervical fractures; at least one of the cervical fractures was managed nonoperatively. Outcomes were assessed along with the optimal follow-up duration and frequency of various diagnostic studies. Results: An average of four follow-up visits occurred on average 28-, 66-, 94-, and 158-day post hospital discharge. The average time between discharge and orthotic clearance was 70 days (±32.4 days). Radiographic studies were obtained at follow-up visits, respectively, in, 82.6%, 83.6%, 94.7%, and 100% of patients. Conclusion: The trend for managing nonoperative cervical fractures was an unduly prolonged follow-up duration that included obtaining too many imaging studies.

Bone Marrow-Derived Stem Cell Factor Regulates Prostate Cancer-Induced Shifts in Pre-Metastatic Niche Composition.

Skeletal metastasis is the leading cause of morbidity and mortality in prostate cancer, with 80% of advanced prostate cancer patients developing bone metastases. Before metastasis, bone remodeling occurs, stimulating pre-metastatic niche formation and bone turnover, and platelets govern this process. Stem cell factor (SCF, Kit Ligand) is increased in advanced prostate cancer patient platelet releasates. Further, SCF and its receptor, CD117/c-kit, correlate with metastatic prostate cancer severity. We hypothesized that bone-derived SCF plays an important role in prostate cancer tumor communication with the bone inducing pre-metastatic niche formation. We generated two cell-specific SCF knockout mouse models deleting SCF in either mature osteoblasts or megakaryocytes and platelets. Using two syngeneic androgen-insensitive murine prostate cancer cell lines, RM1 (Ras and Myc co-activation) and mPC3 (Pten and Trp53 deletion), we examined the role of bone marrow-derived SCF in primary tumor growth and bone microenvironment alterations. Platelet-derived SCF was required for mPC3, but not RM1, tumor growth, while osteoblast-derived SCF played no role in tumor size in either cell line. While exogenous SCF induced proangiogenic protein secretion by RM1 and mPC3 prostate cancer cells, no significant changes in tumor angiogenesis were measured by immunohistochemistry. Like our previous studies, tumor-induced bone formation occurred in mice bearing RM1 or mPC3 neoplasms, demonstrated by bone histomorphometry. RM1 tumor-bearing osteoblast SCF knockout mice did not display tumor-induced bone formation. Bone stromal cell composition analysis by flow cytometry showed significant shifts in hematopoietic stem cell (HSC), mesenchymal stem cell (MSC), and osteoblast cell percentages in mice bearing RM1 or mPC3 tumors. There were no significant changes in the percentage of macrophages, osteoclasts, or osteocytes. Our study demonstrates that megakaryocyte/platelet-derived SCF regulates primary mPC3 tumor growth, while SCF originating from osteoblasts plays a role in bone marrow-derived progenitor cell composition and pre-metastatic niche formation. Further, we show that both the source of SCF and the genetic profile of prostate cancer determine the effects of SCF. Thus, targeting the SCF/CD117 signaling axis with tyrosine kinase inhibitors could affect primary prostate carcinomas or play a role in reducing bone metastasis dependent on the gene deletions or mutations driving the patients' prostate cancer.

Association of Sickle Cell Pain & Symptoms on Health-Related Quality of Life Among Pediatric Patients.

CONTEXT: Sickle cell disease (SCD) may cause significant complications leading to increased disease burden and poor psychosocial functioning. Yet, little is known about specific disease-related variables associated with decreased health-related quality of life (HRQOL) of this population. OBJECTIVES: This study aims to discover whether pain and disease-related symptoms are associated with the HRQOL of pediatric patients diagnosed with SCD. METHODS: A total of 150 patient (ages 8-17 years) and parent dyads were enrolled in this cross-sectional quantitative research study. Measures of HRQOL were gathered using the Pediatric Quality of Life (PedsQL) 3.0 SCD module, while the frequency of pain and SCD-related symptoms was reported by parent-proxy. Linear regression was deployed to analyze whether pain and SCD-related symptoms separately predict HRQOL scores in youth with SCD. RESULTS: Study results revealed that the frequency of pain (P < 0.001) and SCD-related symptoms (P < 0.001) predicted lower HRQOL scores in children and adolescents with SCD. Moreover, age (P < 0.05) emerged as a significant predictor of HRQOL in this sample. CONCLUSION: Pain and disease-related symptoms separately predicted lower HRQOL scores in the study sample, which shows that other SCD-related symptoms aside from pain has the potential to negatively impact youth with SCD. Implications for these findings extend the knowledge of clinicians in the healthcare setting to recognize the detriment of other SCD-related symptoms that may be overlooked, as pain is often the focus.

Further validation of the Perceptions of Uncertainties in Genome Sequencing scale among patients with cancer undergoing tumor sequencing.

It is important to understand how individuals perceive uncertainties and the consequent impact on their psychological well-being and health behavior. The Perceptions of Uncertainties in Genome Sequencing (PUGS) scale measures clinical, affective, and evaluative uncertainties about information from sequencing. The PUGS scale has been shown to be valid and reliable among individuals receiving results about their genomes. This study assessed whether its validity generalized to patients with cancer undergoing tumor sequencing. Exploratory factor analysis (EFA) was conducted on data from the Molecular Screening and Therapeutics Program (n = 310) to identify a measurement model. Confirmatory factor analysis (CFA) was used to determine the adequacy of the resulting fit. EFA identified the same three-factor structure reported previously. CFA confirmed that the measurement model yielded a good fit (χ2 /df = 3.72, CFI = 0.96, SRMR = 0.05, and RMSEA = 0.09) and satisfied convergent and discriminant validity. These findings provide further evidence of the validity and reliability of the PUGS scale in measuring three types of uncertainty. Continued application will facilitate an evidence-based approach to intervention and enhance understanding of what it is like to receive results. In turn, this will improve clinical outcomes as undergoing sequencing becomes an increasingly common experience.

Feeding a high-energy finishing diet upon arrival to high-risk feedlot calves: effects on health, performance, ruminal pH, rumination, serum metabolites, and carcass traits.

This study evaluated the impacts of feeding a high-energy finishing diet during both the receiving and finishing period compared with a lower-energy receiving diet with adaptation to the finishing diet on health, performance, serum chemistry, ruminal pH, rumination, and carcass characteristics of high-risk feedlot cattle. Five truck-load blocks of steers (n = 101) and bulls (n = 299) were used in a generalized complete block design and randomly assigned to receive: 1) finishing diet for the entire feeding period (FIN) or 2) receiving diet for the first 56 d, followed by a transition to the finishing diet (REC). All cattle were fed ad libitum and consumed the same diet by day 74. A subset of cattle (n = 48) was randomly selected to quantify ruminal pH, temperature, and rumination time. Ultrasound images were collected on days 0, 74, and 146 to determine fat thickness over the 12th rib and rump, and carcass characteristics were determined after slaughter. Cattle fed FIN had less (P < 0.01) dry matter intake (DMI) from days 0 to 74, but DMI did not differ (P = 0.80) after day 74. From days 0 to final, DMI was 0.26 kg less for FIN compared with REC (P = 0.01). However, calculated metabolizable energy intake was not different from days 0 to 74 (P = 0.19), days 74 to final (P = 0.80), or overall (P = 0.78). Body weight (BW) on day 74 was greater (P < 0.01) and final BW tended to be greater (P = 0.10) for FIN compared with REC. Cattle consuming FIN had greater (P < 0.01) average daily gain and increased (P < 0.01) gain:feed from days 0 to 74. There were no differences (P ≥ 0.31) in health outcomes. On day 74, FIN had greater (P = 0.04) fat thickness over the rump and rib but did not differ (P ≥ 0.52) on day 146. Carcasses of FIN had greater (P = 0.04) hot carcass weight with no difference (P ≥ 0.11) in ribeye area, 12th rib fat thickness, yield grade, or quality grade. There was no difference (P = 0.18) in liver abscess rate. There was a diet × day interaction for blood urea nitrogen (P = 0.02) such that concentration decreased from days 0 to 28 in both treatments, but was less on day 28 for FIN. Ruminal pH was greater on days 2 and 61 and rumination time was less from days 0 to 28 for FIN (diet × day interaction; P < 0.01). Overall, these results suggest that providing a finishing diet fed ad libitum to high-risk calves upon arrival may be a viable alternative to a low-energy receiving diet.

When high-risk cattle arrive at the feedlot, they have low feed consumption and a greater risk for disease because of stress, inflammation, and exposure to pathogens. Because of reduced feed consumption, newly arrived cattle may not be able to meet their energy requirement for growth during the first several weeks after feedlot arrival. Therefore, providing a high-energy finishing diet (FIN) when stressed cattle arrive at the feedlot may allow for greater growth performance and improved health when compared with a traditional, low-energy receiving diet that contains more roughage (REC). Our study evaluated this concept and we observed that cattle fed FIN had greater body weight, average daily gain, and gain:feed (G:F) within the first 56 d when the different diets were fed, with no difference in growth performance after the cattle consuming REC transitioned to FIN on day 74. However, cattle consuming FIN had greater hot carcass weight and G:F over the entire feeding period. There were no differences in health outcomes among treatments. Overall, providing a high-energy finishing diet to high-risk cattle upon arrival to the feedlot improved growth performance with no impact on health.

Healthcare utilization and the quality of life of children and adolescents with sickle cell disease.

BACKGROUND/OBJECTIVE: Youth diagnosed with sickle cell disease (SCD) are at increased risk of poor health-related quality of life (HRQOL) due to the complexities associated with this disease. The literature notes that predictors such as pain and poor mental health are associated with increased healthcare access; however, the connection between healthcare use and their overall well-being has been understudied. This study investigates whether healthcare utilization predicts the HRQOL in youth with SCD. DESIGN/METHODS: Patients completed the Pediatric Quality of Life (PedsQL) 3.0 SCD module, whereas the researcher conducted a retrospective chart review to gather patient characteristics such as emergency room (ER) and hospitalization occurrences over the past 12 months. RESULTS: The study consisted of 150 pediatric patients with SCD, ages 8-17 years old, and their parents. Patients with ≥ 4 ER visits and hospitalizations reported worse HRQOL scores than their respective counterparts. Additionally, a higher frequency of ER visits (P = 0.05) and hospitalizations (P = 0.005) predicted lower HRQOL scores. Age (P = 0.04) also emerged as a significant predictor for both regression models, as increased healthcare access among older patients with SCD was associated with poorer HRQOL. CONCLUSION: This study found that as youth with SCD require ER treatment and/or hospital admission, they are at increased risk for lower HRQOL, specifically as they get older. Findings suggest that attention should be paid to patients who require more frequent healthcare intervention. Improvement in outpatient care of pediatric patients with SCD may help to mitigate ER and inpatient use.

A systematic review of amino acid PET in assessing treatment response to temozolomide in glioma.

The response assessment in neuro-oncology (RANO) criteria have been the gold standard for monitoring treatment response in glioblastoma (GBM) and differentiating tumor progression from pseudoprogression. While the RANO criteria have played a key role in detecting early tumor progression, their ability to identify pseudoprogression is limited by post-treatment damage to the blood-brain barrier (BBB), which often leads to contrast enhancement on MRI and correlates poorly to tumor status. Amino acid positron emission tomography (AA PET) is a rapidly growing imaging modality in neuro-oncology. While contrast-enhanced MRI relies on leaky vascularity or a compromised BBB for delivery of contrast agents, amino acid tracers can cross the BBB, making AA PET particularly well-suited for monitoring treatment response and diagnosing pseudoprogression. The authors performed a systematic review of PubMed, MEDLINE, and Embase through December 2021 with the search terms "temozolomide" OR "Temodar," "glioma" OR "glioblastoma," "PET," and "amino acid." There were 19 studies meeting inclusion criteria. Thirteen studies utilized [18F]FET, five utilized [11C]MET, and one utilized both. All studies used static AA PET parameters to evaluate TMZ treatment in glioma patients, with nine using dynamic tracer parameters in addition. Throughout these studies, AA PET demonstrated utility in TMZ treatment monitoring and predicting patient survival.

GM-CSF Promotes Immune Response and Survival in a Mouse Model of COVID-19.

COVID-19 results in increased expression of inflammatory cytokines, but inflammation-targeting clinical trials have yielded poor to mixed results. Our studies of other disorders with an inflammatory component, including Alzheimer's disease, chemobrain, Down syndrome, normal aging, and West Nile Virus infection, showed that treatment with the 'pro-inflammatory' cytokine granulocyte-macrophage colony stimulating factor (GM-CSF) in humans or mouse models alleviated clinical, behavioral, and pathological features. We proposed that human recombinant GM-CSF (sargramostim) be repurposed to promote both the innate and adaptive immune responses in COVID-19 to reduce viral load and mortality1. Here, we report the results of a placebo-controlled study of GM-CSF in human ACE2 transgenic mice inoculated intranasally with SARS-CoV2 virus, a model of COVID-19. Infection resulted in high viral titers in lungs and brains and over 85% mortality. GM-CSF treatment beginning one day after infection increased anti-viral antibody titers, lowered mean lung viral titers proportionately (p=0.0020) and increased the odds of long-term survival by up to 5.8-fold (p=0.0358), compared to placebo. These findings suggest that, as an activator of both the innate and adaptive immune systems, GM-CSF/sargramostim may be an effective COVID-19 therapy with the potential to protect from re-infection more effectively than treatment with antiviral drugs or monoclonal antibodies.

Identification of a Novel Delta Opioid Receptor Agonist Chemotype with Potential Negative Allosteric Modulator Capabilities.

The δ-opioid receptor (δOR) holds great potential as a therapeutic target. Yet, clinical drug development, which has focused on δOR agonists that mimic the potent and selective tool compound SNC80 have largely failed. It has increasingly become apparent that the SNC80 scaffold carries with it potent and efficacious ß-arrestin recruitment. Here, we screened a relatively small (5120 molecules) physical drug library to identify δOR agonists that underrecruit ß-arrestin, as it has been suggested that compounds that efficaciously recruit ß-arrestin are proconvulsant. The screen identified a hit compound and further characterization using cellular binding and signaling assays revealed that this molecule (R995045, compound 1) exhibited ten-fold selectivity over µ- and κ-opioid receptors. Compound 1 represents a novel chemotype at the δOR. A subsequent characterization of fourteen analogs of compound 1, however did not identify a more potent δOR agonist. Computational modeling and in vitro characterization of compound 1 in the presence of the endogenous agonist leu-enkephalin suggest compound 1 may also bind allosterically and negatively modulate the potency of Leu-enkephalin to inhibit cAMP, acting as a 'NAM-agonist' in this assay. The potential physiological utility of such a class of compounds will need to be assessed in future in vivo assays.

Site fidelity and behavioral plasticity regulate an ungulate's response to extreme disturbance.

With rapid global change, the frequency and severity of extreme disturbance events are increasing worldwide. The ability of animal populations to survive these stochastic events depends on how individual animals respond to their altered environments, yet our understanding of the immediate and short-term behavioral responses of animals to acute disturbances remains poor. We focused on animal behavioral responses to the environmental disturbance created by megafire. Specifically, we explored the effects of the 2018 Mendocino Complex Fire in northern California, USA, on the behavior and body condition of black-tailed deer (Odocoileus hemionus columbianus). We predicted that deer would be displaced by the disturbance or experience high mortality post-fire if they stayed in the burn area. We used data from GPS collars on 18 individual deer to quantify patterns of home range use, movement, and habitat selection before and after the fire. We assessed changes in body condition using images from a camera trap grid. The fire burned through half of the study area, facilitating a comparison between deer in burned and unburned areas. Despite a dramatic reduction in vegetation in burned areas, deer showed high site fidelity to pre-fire home ranges, returning within hours of the fire. However, mean home range size doubled after the fire and corresponded to increased daily activity in a severely resource-depleted environment. Within their home ranges, deer also selected strongly for patches of surviving vegetation and woodland habitat, as these areas provided forage and cover in an otherwise desolate landscape. Deer body condition significantly decreased after the fire, likely as a result of a reduction in forage within their home ranges, but all collared deer survived for the duration of the study. Understanding the ways in which large mammals respond to disturbances such as wildfire is increasingly important as the extent and severity of such events increases across the world. While many animals are adapted to disturbance regimes, species that exhibit high site fidelity or otherwise fixed behavioral strategies may struggle to cope with increased climate instability and associated extreme disturbance events.