Influence of Sex and Gender on Musculoskeletal Conditions and How They Are Reported.

ABSTRACT: There is increasing evidence that musculoskeletal tissues are differentiallys regulated by sex hormones in males and females. The influence of sex hormones, in addition to other sex-based differences such as in anatomical alignment and immune-system function, impact the prevalence and severity of disease as well as the types of injuries that affect the musculoskeletal system and the outcomes of prevention measures and treatment. Literature specifically addressing sex differences related to the musculoskeletal system is limited, underscoring the imperative for both basic and clinical research on this topic. This review highlights areas of research that have implications for bone and cartilage health, including growth and development, sports injuries, osteoarthritis, osteoporosis, and bone frailty. It is clear that important aspects of the musculoskeletal system have been understudied. Consideration of how sex hormone therapy will affect musculoskeletal tissues in prepuberty, during puberty, and in adults is vital, yet little is known. The purpose of this article is to foster awareness and interest in advancing our understanding of how sex differences influence orthopaedic practice.

Immunologic Predictors of Vaccine Responsiveness in Patients With Lymphoma and Chronic Lymphocytic Leukemia.

Patients with B-cell lymphomas have altered cellular components of vaccine responses due to malignancy and therapy, and the optimal timing of vaccination relative to therapy remains unknown. Severe acute respiratory syndrome coronavirus 2 vaccines created an opportunity for new insights in vaccine timing because patients were challenged with a novel antigen across multiple phases of treatment. We studied serologic messenger RNA vaccine response in retrospective and prospective cohorts with lymphoma and chronic lymphocytic leukemia, paired with clinical and research immune parameters. Reduced serologic response was observed more frequently during active treatment, but nonresponse was also common within observation and posttreatment groups. Total immunoglobulin A and immunoglobulin M correlated with successful vaccine response. In individuals treated with anti-CD19-directed chimeric antigen receptor-modified T cells, nonresponse was associated with reduced B and T follicular helper cells. Predictors of vaccine response varied by disease and therapeutic group, and therefore further studies of immune health during and after cancer therapies are needed to individualize vaccine timing.

Epithelial IL-2 is critical for NK cell-mediated cancer immunosurveillance in mammary glands.

Interleukin 2 (IL-2) is the first identified cytokine and its interaction with receptors has been known to shape the immune responses in many lymphoid or non-lymphoid tissues for more than four decades. Active T cells are the primary cellular source for IL-2 production and epithelial cells have never been considered the major cellular source of IL-2 under physiological conditions. It is, however, tempting to speculate that epithelial cells could potentially express IL-2 that regulates the intricate interactions between epithelial cells and lymphocytes. Datamining our recently published single-cell RNAseq in the mouse mammary gland identified IL-2 expression in mammary epithelial cells, which is induced by prolactin via the STAT5 signaling pathway. Furthermore, epithelial IL-2 plays a crucial role in maintaining the physiological functions of natural killer (NK) cells within the mammary glands. IL-2 deletion in the mammary epithelial cells leads to a significant reduction in the number and function of NK cells, which in turn results in defective immunosurveillance, expansion of luminal epithelial cells, and tumor development. Interestingly, T cells in the mammary glands are not changed, indicating the specific regulation of NK cells by epithelial IL-2 production. In agreement, we also found that human epithelial cells express IL-2 and NK cells express the highest level of IL2RB among all the immune cells. Here, we provide the first evidence that epithelial cells produce IL-2, which is critical for maintaining the physiological functions of NK cells in immunosurveillance.

Surgical procedure of intratympanic injection and inner ear pharmacokinetics simulation in domestic pigs.

Introduction: One major obstacle in validating drugs for the treatment or prevention of hearing loss is the limited data available on the distribution and concentration of drugs in the human inner ear. Although small animal models offer some insights into inner ear pharmacokinetics, their smaller organ size and different barrier (round window membrane) permeabilities compared to humans can complicate study interpretation. Therefore, developing a reliable large animal model for inner ear drug delivery is crucial. The inner and middle ear anatomy of domestic pigs closely resembles that of humans, making them promising candidates for studying inner ear pharmacokinetics. However, unlike humans, the anatomical orientation and tortuosity of the porcine external ear canal frustrates local drug delivery to the inner ear. Methods: In this study, we developed a surgical technique to access the tympanic membrane of pigs. To assess hearing pre- and post-surgery, auditory brainstem responses to click and pure tones were measured. Additionally, we performed 3D segmentation of the porcine inner ear images and used this data to simulate the diffusion of dexamethasone within the inner ear through fluid simulation software (FluidSim). Results: We have successfully delivered dexamethasone and dexamethasone sodium phosphate to the porcine inner ear via the intratympanic injection. The recorded auditory brainstem measurements revealed no adverse effects on hearing thresholds attributable to the surgery. We have also simulated the diffusion rates for dexamethasone and dexamethasone sodium phosphate into the porcine inner ear and confirmed the accuracy of the simulations using in-vivo data. Discussion: We have developed and characterized a method for conducting pharmacokinetic studies of the inner ear using pigs. This animal model closely mirrors the size of the human cochlea and the thickness of its barriers. The diffusion time and drug concentrations we reported align closely with the limited data available from human studies. Therefore, we have demonstrated the potential of using pigs as a large animal model for studying inner ear pharmacokinetics.

Online protein digestion in membranes between capillary electrophoresis and mass spectrometry.

This research employs pepsin-containing membranes to digest proteins online after a capillary electrophoresis (CE) separation and prior to tandem mass spectrometry. Proteolysis after the separation allows the peptides from a given protein to enter the mass spectrometer in a single plug. Thus, migration time can serve as an additional criterion for confirming the identification of a peptide. The membrane resides in a sheath-flow electrospray ionization (ESI) source to enable digestion immediately before spray into the mass spectrometer, thus limiting separation of the digested peptides. Using the same membrane, digestion occurred reproducibly during 20 consecutive CE analyses performed over a 10 h period. Additionally, after separating a mixture of six unreduced proteins with CE, online digestion facilitated protein identification with at least 2 identifiable peptides for all the proteins. Sequence coverages were >75% for myoglobin and carbonic anhydrase II but much lower for proteins containing disulfide bonds. Development of methods for efficient separation of reduced proteins or identification of cross-linked peptides should enhance sequence coverages for proteins with disulfide bonds. Migration times for the peptides identified from a specific protein differed by <∼30 s, which allows for rejection of some spurious peptide identifications.

The unstructured linker of Mlh1 contains a motif required for endonuclease function which is mutated in cancers.

Eukaryotic DNA mismatch repair (MMR) depends on recruitment of the Mlh1-Pms1 endonuclease (human MLH1-PMS2) to mispaired DNA. Both Mlh1 and Pms1 contain a long unstructured linker that connects the N- and carboxyl-terminal domains. Here, we demonstrated the Mlh1 linker contains a conserved motif (Saccharomyces cerevisiae residues 391-415) required for MMR. The Mlh1-R401A,D403A-Pms1 linker motif mutant protein was defective for MMR and endonuclease activity in vitro, even though the conserved motif could be >750 Å from the carboxyl-terminal endonuclease active site or the N-terminal adenosine triphosphate (ATP)-binding site. Peptides encoding this motif inhibited wild-type Mlh1-Pms1 endonuclease activity. The motif functioned in vivo at different sites within the Mlh1 linker and within the Pms1 linker. Motif mutations in human cancers caused a loss-of-function phenotype when modeled in S. cerevisiae. These results suggest that the Mlh1 motif promotes the PCNA-activated endonuclease activity of Mlh1-Pms1 via interactions with DNA, PCNA, RFC, or other domains of the Mlh1-Pms1 complex.

PD-1 combination therapy with IL-2 modifies CD8+ T cell exhaustion program.

Combination therapy with PD-1 blockade and IL-2 is highly effective during chronic lymphocytic choriomeningitis virus infection1. Here we examine the underlying basis for this synergy. We show that PD-1 + IL-2 combination therapy, in contrast to PD-1 monotherapy, substantially changes the differentiation program of the PD-1+TCF1+ stem-like CD8+ T cells and results in the generation of transcriptionally and epigenetically distinct effector CD8+ T cells that resemble highly functional effector CD8+ T cells seen after an acute viral infection. The generation of these qualitatively superior CD8+ T cells that mediate viral control underlies the synergy between PD-1 and IL-2. Our results show that the PD-1+TCF1+ stem-like CD8+ T cells, also referred to as precursors of exhausted CD8+ T cells, are not fate-locked into the exhaustion program and their differentiation trajectory can be changed by IL-2 signals. These virus-specific effector CD8+ T cells emerging from the stem-like CD8+ T cells after combination therapy expressed increased levels of the high-affinity IL-2 trimeric (CD25-CD122-CD132) receptor. This was not seen after PD-1 blockade alone. Finally, we show that CD25 engagement with IL-2 has an important role in the observed synergy between IL-2 cytokine and PD-1 blockade. Either blocking CD25 with an antibody or using a mutated version of IL-2 that does not bind to CD25 but still binds to CD122 and CD132 almost completely abrogated the synergistic effects observed after PD-1 + IL-2 combination therapy. There is considerable interest in PD-1 + IL-2 combination therapy for patients with cancer2,3, and our fundamental studies defining the underlying mechanisms of how IL-2 synergizes with PD-1 blockade should inform these human translational studies.

Examining Relationships Between Groundwater Nitrate Concentrations in Drinking Water and Landscape Characteristics to Understand Health Risks.

Nitrate ingested from drinking water has been linked to adverse health outcomes (e.g., cancer, birth defects) at levels as low as ∼2 mg/L NO3-N, far below the regulatory limits of 10 mg/L. In many areas, groundwater is a common drinking water source and may contain elevated nitrate, but limited data on the patterns and concentrations are available. Using an extensive regulatory data set of over 100,000 nitrate drinking water well samples, we developed new maps of groundwater nitrate concentrations from 76,724 wells in Michigan's Lower Peninsula, USA for the 2006-2015 period. Kriging, a geostatistical method, was used to interpolate concentrations and quantify probability of exceeding relevant thresholds (>0.4 [common detection limit], >2 mg/L NO3-N). We summarized this probability in small watersheds (∼80 km2) to identify correlated variables using the machine learning method classification and regression trees (CARTs). We found 79% of wells had concentrations below the detection limit in this analysis (<0.4 mg/L NO3-N). In the shallow aquifer (focus of study), 13% of wells exceeded 2 mg/L NO3-N and 2% exceeded the EPA maximum contaminant level of 10 mg/L. CART explained 40%-45% of variation in each model and identified three categories of critical correlated variables: source (high agricultural nitrogen inputs), vulnerable soil conditions (low soil organic carbon and high hydraulic conductivity), and transport mechanisms (high aquifer recharge). These findings add to the body of literature seeking to identify communities at risk of elevated nitrate and study associated adverse health outcomes.

Social Media Use by Hand Surgery Fellowship Programs.

INTRODUCTION: Social media has emerged as a useful tool in the fellowship recruitment process. We aimed to assess the prevalence of social media use among hand surgery fellowships, to analyze social media posts according to content, and to evaluate the level of engagement generated by specific content. METHODS: We used a list of accredited hand surgery fellowships from the American Society for Surgery of the Hand Fellowship Directory to identify all hand surgery fellowship profiles on Facebook, Twitter, and Instagram. Instagram was the most commonly used platform and thus the focus of this study. Two reviewers independently assessed all Instagram posts from each program and assigned content labels. We assessed the variability in content published by each program using a Monte Carlo estimation of an exact chi-square test. We calculated the level of engagement generated by each content label using the number of likes per post per number of account followers. We analyzed the variability in engagement using a Kruskal-Wallis test. RESULTS: We identified 21 Instagram accounts from 89 fellowship programs (24%). Seventeen of 21 (81%) were created after the onset of the coronavirus disease 2019 pandemic. There was significant variability in the scope of content published by each program ( P < 0.0001) and in the level of engagement generated by each content label ( P < 0.0001). Skills, conferences, fellow, case example(s), faculty, and team dynamics generated some of the most engagement. Logistics, miscellaneous, and facilities generated the least. DISCUSSION: There is wide variability in the content produced by hand fellowship programs. Specific types of content generate more engagement from followers than others. This information may guide fellowship programs to produce the type of content potential applicants find most useful when making application and rank list decisions.

Clinical Pathways of Patients Denied Total Knee Arthroplasty Due to an Institutional BMI Cutoff.

Out of concern for the increased risk of complications with morbid obesity, institutional body mass index (BMI) cutoffs for total knee arthroplasty (TKA) have become commonplace. We sought to answer the questions: what percentage of morbidly obese patients with knee osteoarthritis who present to an arthroplasty clinic will, within 2 years, undergo TKA at (1) a BMI less than 40 kg/m2 or (2) at a BMI greater than 40 kg/m2? Of those who do not undergo surgery, (3) what percentage lose enough weight to become TKA-eligible, and (4) what percentage do not? We performed an observational study of 288 patients, of which 256 had complete follow-up. Institutional electronic medical record review and patient follow-up by telephone were conducted to determine which patients underwent surgery, and at what BMI. For those that did not undergo TKA, BMI was examined to see if the patient ever lost enough weight to become TKA eligible. Twelve of 256 patients (4.7%) underwent TKA at a BMI less than 40 kg/m2, 64 patients (25%) underwent TKA at a BMI greater than 40 kg/m2, and 7 patients (2.7%) underwent surgery at an outside hospital. The average BMI at the time of surgery was 42.3 kg/m2. Thirty-seven of 256 patients (14.4%) lost enough weight to become TKA-eligible within 2 years of the initial visit but did not undergo surgery, while 136 patients (53.1%) neither underwent TKA nor became eligible. Strict enforcement of a BMI cutoff for TKA is variable among surgeons. In the absence of weight loss protocols, 19.1% of morbidly obese patients may be expected to reach the sub-40 kg/m2 BMI milestone.

Rad27 and Exo1 function in different excision pathways for mismatch repair in Saccharomyces cerevisiae.

Eukaryotic DNA Mismatch Repair (MMR) involves redundant exonuclease 1 (Exo1)-dependent and Exo1-independent pathways, of which the Exo1-independent pathway(s) is not well understood. The exo1Δ440-702 mutation, which deletes the MutS Homolog 2 (Msh2) and MutL Homolog 1 (Mlh1) interacting peptides (SHIP and MIP boxes, respectively), eliminates the Exo1 MMR functions but is not lethal in combination with rad27Δ mutations. Analyzing the effect of different combinations of the exo1Δ440-702 mutation, a rad27Δ mutation and the pms1-A99V mutation, which inactivates an Exo1-independent MMR pathway, demonstrated that each of these mutations inactivates a different MMR pathway. Furthermore, it was possible to reconstitute a Rad27- and Msh2-Msh6-dependent MMR reaction in vitro using a mispaired DNA substrate and other MMR proteins. Our results demonstrate Rad27 defines an Exo1-independent eukaryotic MMR pathway that is redundant with at least two other MMR pathways.

Rapid induction of antigen-specific CD4+ T cells is associated with coordinated humoral and cellular immunity to SARS-CoV-2 mRNA vaccination.

SARS-CoV-2 mRNA vaccines have shown remarkable clinical efficacy, but questions remain about the nature and kinetics of T cell priming. We performed longitudinal antigen-specific T cell analyses on healthy SARS-CoV-2-naive and recovered individuals prior to and following mRNA prime and boost vaccination. Vaccination induced rapid antigen-specific CD4+ T cell responses in naive subjects after the first dose, whereas CD8+ T cell responses developed gradually and were variable in magnitude. Vaccine-induced Th1 and Tfh cell responses following the first dose correlated with post-boost CD8+ T cells and neutralizing antibodies, respectively. Integrated analysis revealed coordinated immune responses with distinct trajectories in SARS-CoV-2-naive and recovered individuals. Last, whereas booster vaccination improved T cell responses in SARS-CoV-2-naive subjects, the second dose had little effect in SARS-CoV-2-recovered individuals. These findings highlight the role of rapidly primed CD4+ T cells in coordinating responses to the second vaccine dose in SARS-CoV-2-naive individuals.

Adoption, Sustainability, and Dissemination of Chronic Disease Prevention Policies in Community-Based Organizations.

Introduction. Despite increasing interest in structural (policy, systems, and environmental) changes to improve health, little attention has focused on the adoption, implementation, sustainability, and potential for dissemination of these changes among local community-based organizations. Method. A mixed methods approach was used for this process evaluation. Representatives of nine community-based organizations were surveyed using closed-ended questions and in-depth qualitative interviews to describe 32 policy changes. Diffusion of Innovation theory was used to inform the development of survey questions and the interview guide. Results. Policies adopted by local community-based organizations concerned types of food/beverages provided to staff/clients, methods to encourage physical activity, breastfeeding support, and tobacco control. The majority of the policies were either fully (66%) or partially (31%) implemented 1 year after their initial adoption. In general, participants somewhat/strongly agreed that policies had characteristics that predict sustainability/diffusion (relative advantage, compatibility, complexity, trialability, observability). In-depth interview responses described a generally smooth process for policy adoption and high levels of optimism for continued sustainability but revealed few efforts to disseminate the policies beyond the original organization. Conclusions. Structural changes in community-based organizations are a valuable tool for encouraging healthy changes in communities and have great potential to be adopted, sustained, and diffused.

Gleason Score Evolution and the Effect on Prostate Cancer Outcomes.

OBJECTIVES: We evaluated how the changes in Gleason grading affected the long-term outcomes of a large prostatectomy cohort. METHODS: We obtained long-term follow-up (16.7 years) in 581 patients having undergone radical retropubic prostatectomy between 1985 and 1995. We excluded those with seminal vesicle and/or lymphatic involvement. We regraded the specimens according to contemporary guidelines and compared how this affected outcomes compared with their original (pre-1995) Gleason scoring. In total, 499 patients were evaluable. RESULTS: A Gleason score of 6 or less declined from 73% to 29%, and the number increased from 25% to 63% for a Gleason score of 7 and from 5% to 8% for a Gleason score of 8 to 9. As a result, for a Gleason score less than 7, biochemical failure decreased from 28% to 23%, metastatic disease 5% to 2%, and prostate cancer death from 5% to 3%. The same results were 50% to 37%, 11% to 7%, and 10% to 6% for a Gleason score of 7 and 86% to 71%, 43% to 32%, and 29% to 26% for a Gleason score more than 7, respectively. With the most recent grade grouping, for groups 1 to 5, biochemical failure occurred in 23%, 32%, 45%, 69%, and 78%, respectively. Metastatic disease occurred in 2%, 4%, 12%, 24%, and 56%, respectively. Prostate cancer-related death occurred in 2%, 4%, 9%, 21%, and 44%, respectively. CONCLUSIONS: The revised Gleason scores improved the outcomes in all risk groups. Based on Gleason score, patients with prostate cancer will appear to have better outcomes than they did before 2005, making any comparison tenable. The current grading system shows a consistent increased risk in biochemical failure, metastatic disease, and prostate cancer-related death with each successive grade.

Predictive value of lateral soft tissue thickness for complications after total hip arthroplasty with a lateral incision.

The purpose of this study was to determine the relationship between soft tissue thickness lateral to the greater trochanter, as measured on anteroposterior pelvis radiograph, and postoperative complications following primary total hip arthroplasty. A retrospective review of 1110 consecutive patients treated at a single institution from 2003 to 2011 was conducted. Postoperative complications were divided into surgical site infections, deep wound infections, noninfectious surgical complications, need for revision surgery, and medical complications. Lateral soft tissue thickness (LSTT) was measured as the horizontal distance from the most lateral point on the greater trochanter to the skin edge obtained from anteroposterior hip radiographs. Among the 1110 study patients, 19.19% had a postoperative complication, with a deep infection rate of 3.42%. Of the previously identified risk factors, increased LSTT and body mass index were both associated with surgical site infection and deep infection, and LSTT was associated with revision surgery. An LSTT value of >5 cm was predictive of surgical site infection, deep infection, and revision surgery. This easily obtainable radiographic measurement, along with clinical examination near the operative site, might prove helpful in making preoperative risk assessments.

In Utero Restoration of Hindbrain Herniation in Fetal Myelomeningocele as Part of Prenatal Regenerative Therapy Program at Mayo Clinic.

OBJECTIVE: To assess our initial experience with prenatal restoration of hindbrain herniation following in utero repair of myelomeningocele (MMC). PATIENTS AND METHODS: Three consecutive patients with prenatally diagnosed MMC (between January 1, 2018 and September 30, 2018) were managed with open in utero surgery. As per institutional review board approval and following a protocol designed at the Mayo Clinic Maternal & Fetal Center, fetal intervention was offered between 19 0/7 and 25 6/7 weeks of gestation. Prenatal improvement of hindbrain herniation was the declared restorative end point. Obstetrical and perinatal outcomes were also assessed. RESULTS: Diagnosis of MMC was confirmed upon referral between 20 and 21 weeks' gestation by using fetal ultrasound and magnetic resonance imaging. In all cases reported here, the spinal defect was lumbosacral with evidence of hindbrain herniation. Open in utero MMC repair was performed between 24 and 25 weeks' gestation with no notable perioperative complications. Postprocedure fetal magnetic resonance imaging performed 6 weeks after in utero repair documented improvement of hindbrain herniation. Deliveries were at 37 weeks by cesarean section without complications. Most recent postnatal follow-ups were unremarkable at both 11 months (baby 1) and 3 months of age (baby 2), with mild ventriculomegaly. Antenatal and postnatal follow-up of baby 3 at 1 month of age was also unremarkable. CONCLUSION: Our study highlights the prenatal restoration of hindbrain herniation following in utero MMC repair in all cases presented here as an example of a prenatal regenerative therapy program in our institution.

Prospective Study of Acute Opioid Use After Adolescent Anterior Cruciate Ligament Reconstruction Shows No Effect From Patient- or Surgical-Related Factors.

INTRODUCTION: Patient-reported pain scores and opioid use have not been quantified after outpatient adolescent anterior cruciate ligament reconstruction (ACLR). METHODS: Patients aged 12 to 18 years undergoing primary isolated ACLR, with or without meniscal treatment, were prospectively recruited. Patients actively taking opioids or with previous extended use of opioids were excluded. Two orthopaedic surgeons performed ACLR and determined the use of a hamstring or bone-patellar tendon-bone autograft. For postoperative pain management, patients were prescribed 40 tablets of hydrocodone/acetaminophen 5/325 mg. Patients were instructed to document daily pill consumption and side effects through a daily log for 6 weeks. Patients completed the American Pain Society Patient Outcome Questionnaire at the end of weeks 1 and 6. RESULTS: One hundred three patients were enrolled, with age: 12.5 to 18.9 years (mean 16.2 y ± 1.3), weight: 41.3 to 113.6 kg (mean 72.4 kg ± 17.2), and body mass index: 17.8 to 40.1 (mean 25.9 ± 4.9). Sixty-nine patients received a hamstring autograft, and 34 received a bone-patellar tendon-bone autograft. Fifty-six received additional meniscal procedures. The median number of postoperative opioids taken by patients was 17 (range 0 to 40). No notable differences were found in total pill consumption with regard to age, weight, body mass index, sex, block type, autograft type, or meniscal treatment at 1 week post-op or 6 weeks post-op. No correlation was found between the self-reported "worst pain in the past 24 hours" at the end of the first postoperative week or after 6 weeks (r = 0.112, P = 0.26, and r = 0.093, P = 0.36). No correlation was found between the level of satisfaction with pain treatment and total number of pills taken during the first postoperative week or at the end of 6 weeks (r = -0.090, P = 0.37, and r = -0.172, P = 0.08). CONCLUSION: Patients take most pain medication during the first postoperative week after adolescent ACLR, although patient and surgical variables had no notable influence on pill consumption. LEVEL OF EVIDENCE: Level IV, case series.

Validating the management paradigm for pediatric spinal aneurysmal bone cysts to optimize long-term outcomes: an institutional experience.

BACKGROUND: The optimal clinical management and outcomes of rare pediatric spinal aneurysmal bone cysts (spABC) is largely anecdotal. Current practice is based on bigger adult series, although given the disparities in spine growth of adults versus children, what impact this difference may have on long-term outcomes has yet to be substantiated. Correspondingly, the aim of this study was to describe the clinical course of all pediatric spABC cases managed at our institution to better understand this. METHODS: A retrospective cohort study of all pediatric spABC cases presenting to our institution between 1993 and 2017 was performed using a predetermined set of selection criteria. Primary outcomes of interest were treatment modalities and their outcomes, recurrence status, and functional status. RESULTS: A total of 24 pediatric spABC cases satisfied all criteria. Median age of diagnosis was 13.5 years, with 15 females and 9 males. Radicular pain was the presenting symptom in 21 (88%) cases. Diagnostic biopsy was pursued in 9 (38%) cases, pre-operative embolization in 8 (33%) cases, surgical intervention in 23 (96%) cases, and sclerotherapy in 2 (8%) cases. In terms of surgery, there were no intraoperative complications, and gross total resection (GTR) was achieved in 14 of the 23 (61%) cases. Overall, there were 5 (21%) cases which experienced recurrence by a median time of 8 months after initial surgery, all of which had initial subtotal resection. Median follow-up was 5 years, by which all patients demonstrated excellent functional status. CONCLUSIONS: There are a number of feasible therapeutic modalities and combinations that can be utilized to maximize control of pediatric spABCs and optimize long-term function similar to that of adults, irrespective of developing versus developed spines. The incidence of recurrence is not negligible, and therefore, rigorous long-term surveillance is highly encouraged, particularly within the first post-operative year following mono-modal non-GTR treatment.

G1T48, an oral selective estrogen receptor degrader, and the CDK4/6 inhibitor lerociclib inhibit tumor growth in animal models of endocrine-resistant breast cancer.

PURPOSE: The combination of targeting the CDK4/6 and estrogen receptor (ER) signaling pathways with palbociclib and fulvestrant is a proven therapeutic strategy for the treatment of ER-positive breast cancer. However, the poor physicochemical properties of fulvestrant require monthly intramuscular injections to patients, which limit the pharmacokinetic and pharmacodynamic activity of the compound. Therefore, an orally available compound that more rapidly reaches steady state may lead to a better clinical response in patients. Here, we report the identification of G1T48, a novel orally bioavailable, non-steroidal small molecule antagonist of ER. METHODS: The pharmacological effects and the antineoplastic mechanism of action of G1T48 on tumors was evaluated using human breast cancer cells (in vitro) and xenograft efficacy models (in vivo). RESULTS: G1T48 is a potent and efficacious inhibitor of estrogen-mediated transcription and proliferation in ER-positive breast cancer cells, similar to the pure antiestrogen fulvestrant. In addition, G1T48 can effectively suppress ER activity in multiple models of endocrine therapy resistance including those harboring ER mutations and growth factor activation. In vivo, G1T48 has robust antitumor activity in a model of estrogen-dependent breast cancer (MCF7) and significantly inhibited the growth of tamoxifen-resistant (TamR), long-term estrogen-deprived (LTED) and patient-derived xenograft tumors with an increased response being observed with the combination of G1T48 and the CDK4/6 inhibitor lerociclib. CONCLUSIONS: These data show that G1T48 has the potential to be an efficacious oral antineoplastic agent in ER-positive breast cancer.

Heat stress impairs in vitro development of preantral follicles of cattle.

Although detrimental effects of heat stress on antral follicle development have been well studied, long-term effects - affecting the preantral follicle pool - are still largely unknown. The goal of this study was to evaluate effects of heat stress on growth, viability, gene expression and ATP production of preantral follicles of cattle. Follicles at the primary, early secondary and secondary stages were isolated from cattle ovaries and individually cultured while imposing physiological (CON; 38.5 °C) or intermittent heat stress (HS; 38.5 °C for 16 h and 41 °C for 8 h daily) conditions for 7 days. Individual follicles were subjected to real-time qPCR for determination of relative abundance of BAX, HSPA1A and SOD1 mRNA transcripts and evaluated for ATP production. Treatment for 7 days with intermittent HS decreased viability (P =  0.01) and diameter (P =  0.03) of preantral follicles. Relative abundances of BAX and HSPA1A mRNA transcripts were greater in follicles of the CON and HS groups that became non-viable during culture (P <  0.05); relative abundance of SOD1 mRNA transcript, however, was only greater in non-viable follicles of the HS group (P <  0.05), but not non-viable follicles of the CON group (P =  0.3). The ATP production was not different between viable follicles of the CON and HS group (P = 0.86). In conclusion, all stages of growing preantral follicles of cattle were susceptible to negative effects of heat stress. Follicles at the secondary stage of development were most sensitive, followed by early secondary and primary follicles.