Next Generation Weight Loss Drugs for the Prevention of Cancer?

Background: Western populations are losing the battle over healthy weight management, and excess body weight is a notable cancer risk factor at the population level. There is ongoing interest in pharmacological interventions aimed at promoting weight loss, including GLP-1 receptor agonists (GLP-1RA), which may be a useful tool to stem the rising tide of obesity-related cancers. Purpose: To investigate the potential of next generation weight loss drugs (NGWLD) like GLP-1RA in population-level chemoprevention.Research Design: We used the OncoSim microsimulation tool to estimate the population-level reductions in obesity and the potentially avoidable obesity-related cancers in Canada over the next 25 years.Results: We estimated a total of 71 281 preventable cancers by 2049, with 36 235 and 35 046 cancers prevented for females and males, respectively. Among the 327 254 total projected cancer cases in 2049, 1.3% are estimated to be preventable through intervention with NGWLD.Conclusions: Pharmacologic intervention is not the ideal solution for the obesity-related cancer crisis. However, these agents and subsequent generations provide an additional tool to rapidly reduce body weight and adiposity in populations that have been extremely challenging to reduce weight with standard diet and exercise approaches. Additional research is needed around approaches to prevent initial weight gain and maintain long-term weight loss.

Differential Expression of LLT1, SLAM Receptors CS1 and 2B4 and NCR Receptors NKp46 and NKp30 in Pediatric Acute Lymphoblastic Leukemia (ALL).

Acute lymphoblastic leukemia (ALL) represents the most common pediatric cancer. Most patients (85%) develop B-cell ALL; however, T-cell ALL tends to be more aggressive. We have previously identified 2B4 (SLAMF4), CS1 (SLAMF7) and LLT1 (CLEC2D) that can activate or inhibit NK cells upon the interaction with their ligands. In this study, the expression of 2B4, CS1, LLT1, NKp30 and NKp46 was determined. The expression profiles of these immune receptors were analyzed in the peripheral blood mononuclear cells of B-ALL and T-ALL subjects by single-cell RNA sequencing data obtained from the St. Jude PeCan data portal that showed increased expression of LLT1 in B-ALL and T-ALL subjects. Whole blood was collected from 42 pediatric ALL subjects at diagnosis and post-induction chemotherapy and 20 healthy subjects, and expression was determined at the mRNA and cell surface protein level. A significant increase in cell surface LLT1 expression in T cells, monocytes and NK cells was observed. Increased expression of CS1 and NKp46 was observed on monocytes of ALL subjects at diagnosis. A decrease of LLT1, 2B4, CS1 and NKp46 on T cells of ALL subjects was also observed post-induction chemotherapy. Furthermore, mRNA data showed altered expression of receptors in ALL subjects pre- and post-induction chemotherapy treatment. The results indicate that the differential expression of the receptors/ligand may play a role in the T-cell- and NK-cell-mediated immune surveillance of pediatric ALL.

Regenerative farming as climate action.

Regenerative agriculture is an alternative approach to farming that has been gaining traction and interest among farmers due to its potential to reduce input costs, improve soil health, and increase the resilience of farming systems. This paper undertakes a practice-based analysis of farmers, applying a lens of regenerative agriculture. Surveys were developed as a part of a broader project using an established methodological framework. Topics were developed and adapted with input from local stakeholders before being mailed out to three farming regions across Australia (the Western Australian Wheatbelt, the Eyre Peninsula in South Australia and Central West New South Wales). The research clustered farmers into two groups: those who are using best-practices that fall inside the scope of regenerative agriculture, and those who are not. The similarities and differences in farmer attributes, as well as self-reported knowledge levels and information sources used by each group are explored. Results indicate that a belief in anthropogenic climate change may be one of the primary divides between the two groups, and therefore a possible driver of best-practice implementation. The findings provide insight into perceptions of regenerative agriculture for Australian farmers, and may assist with knowledge dissemination amongst those managing our environment.

Eatomics: Shiny Exploration of Quantitative Proteomics Data.

Quantitative proteomics data are becoming increasingly more available, and as a consequence are being analyzed and interpreted by a larger group of users. However, many of these users have less programming experience. Furthermore, experimental designs and setups are getting more complicated, especially when tissue biopsies are analyzed. Luckily, the proteomics community has already established some best practices on how to conduct quality control, differential abundance analysis and enrichment analysis. However, an easy-to-use application that wraps together all steps for the exploration and flexible analysis of quantitative proteomics data is not yet available. For Eatomics, we utilize the R Shiny framework to implement carefully chosen parts of established analysis workflows to (i) make them accessible in a user-friendly way, (ii) add a multitude of interactive exploration possibilities, and (iii) develop a unique experimental design setup module, which interactively translates a given research hypothesis into a differential abundance and enrichment analysis formula. In this, we aim to fulfill the needs of a growing group of inexperienced quantitative proteomics data analysts. Eatomics may be tested with demo data directly online via https://we.analyzegenomes.com/now/eatomics/ or with the user's own data by installation from the Github repository at https://github.com/Millchmaedchen/Eatomics.

Novel Magnetic Resonance Imaging Classification of Osteochondritis Dissecans of the Knee: A Reliability Study.

BACKGROUND: Current advanced imaging classification systems for osteochondritis dissecans (OCD) of the knee grade severity of disease by identifying certain lesion characteristics. The most widely used are the Hefti and Nelson systems. A novel classification presents a simpler 3-group approach to diagnose knee OCD by magnetic resonance imaging (MRI), compared with the Hefti (5-group) and Nelson (4-group) classifications. The purpose of this study was to compare the reliability of this novel classification with that of the more complex, established systems-an initial step in establishing validity and clinical utility. METHODS: In total, 120 standardized knee MRIs of patients with established knee OCD were preselected to capture the spectrum of lesion types, with regard to both progression and location of the lesion. Each of the MRIs were independently classified by 2 readers into the novel, Hefti, and Nelson classification systems. A random sample was rereviewed by 1 rater 6 weeks after initial review. The inter-rater and intrarater agreements were evaluated by estimating Krippendorff α. RESULTS: In total, 106 knees were classified by the novel, Hefti, and Nelson classification systems, as 14 of the knees lacked the necessary MRI sequences. There were no differences in inter-rater and intrarater agreement across classification systems. Krippendorff α for inter-rater agreement was 0.51 (95% confidence interval, 0.33-0.66) for the Hefti classification, 0.50 (0.34-0.64) for the Nelson classification, and 0.49 (0.32-0.65) for the novel classification. The intrarater agreement was 0.88 (0.75-0.97) for the Hefti classification, 0.94 (0.86-0.99) for the Nelson classification, and 0.98 (0.94-1.00) for the novel classification system. CONCLUSIONS: The novel classification for knee OCD demonstrated near-perfect intrarater agreement and moderate inter-rater agreement, consistent with the current, well-established classification systems. Pending a subsequent study on validity and clinical utility, this simpler classification system may offer an alternative, noninvasive diagnostic method to guide clinical treatment. LEVEL OF EVIDENCE: Level IV.

Impact of the Microbiome on the Immune System.

Higher organisms are all born with general immunity as well as with, increasingly, more specific immune systems. All immune mechanisms function with the intent of aiding the body in defense against infection. Internal and external factors alike have varying effects on the immune system, and the immune response is tailored specifically to each one. Accompanying the components of the human innate and adaptive immune systems are the other intermingling systems of the human body. Increasing understanding of the body's immune interactions with other systems has opened new avenues of study, including that of the microbiome. The microbiome has become a highly active area of research over the last 10 to 20 years since the NIH began funding the Human Microbiome Project (HMP), which was established in 2007. Several publications have focused on the characterization, functions, and complex interplay of the microbiome as it relates to the rest of the body. A dysfunction between the microbiome and the host has been linked to various diseases including cancers, metabolic deficiencies, autoimmune disorders, and infectious diseases. Further understanding of the microbiome and its interaction with the host in relation to diseases is needed in order to understand the implications of microbiome dysfunction and the possible use of microbiota in the prevention of disease. In this review, we have summarized information on the immune system, the microbiome, the microbiome's interplay with other systems, and the association of the immune system and the microbiome in diseases such as diabetes and colorectal cancer.

The effect of varying degrees of compression from elastic vs plastic wrap on quadriceps intramuscular temperature during wetted ice application.

The aim of this study was to evaluate and compare the effectiveness of wetted ice bag, applied with high compression elastic wrap or held in place with low compression plastic wrap, on reducing vastus lateralis intramuscular temperature and skin surface temperature. Ten healthy male participants had wetted ice packs applied to a standardized area on the anterior aspect of the quadriceps simultaneously to both legs for 30 minutes. The ice pack was secured with high compression (elastic wrap) to the left anterior thigh (60.6 ± 8.1 mm Hg) and low compression (plastic wrap) to the right anterior thigh (15.5 ± 4.0 mm Hg). Intramuscular temperature (1 and 3 cm) and skin temperature of the vastus lateralis were measured continuously during a 10-minute baseline period, 30-minute treatment period, and a 60-minute recovery period. No difference was observed between treatments in terms of the magnitude of reduction in intramuscular temperature at both 1 and 3 cm and skin temperature regardless of compression pressure (P > 0.05). Temperature upon conclusion of elastic wrap treatment was as follows: 17.8 ± 5.2°C at 1 cm and 23.1 ± 4.9°C at 3 cm; plastic wrap treatment: 17.9 ± 4.4°C at 1 cm and 24.5 ± 6.7°C at 3 cm. Plastic wraps may offer a practical alternative to elastic wraps for clinicians as they may be disposed of by the patient or athlete without having to stay at the treatment facility.

Is Anteromedial Drilling Safe in Transphyseal Anterior Cruciate Ligament Reconstruction in Adolescents with Growth Remaining?

BACKGROUND: Previous reports of transphyseal drilling in anterior cruciate ligament (ACL) reconstruction have demonstrated good clinical outcomes without subjective changes in further skeletal development. The purpose of this study is to evaluate radiographic changes during continued growth following a transphyseal ACL reconstruction using an anteromedial femoral (AM) drilling technique in patients with >18 months of growth remaining. METHODS: A review of consecutive adolescents who underwent a soft tissue transphyseal ACL reconstruction using an AM drilling technique was performed. Inclusion criteria was 18 months of growth remaining based on radiographic bone age and standing radiographs at least one year from the index procedure. Demographic, preoperative, and postoperative data, and follow-up three-foot standing lower extremity radiographs were reviewed. Radiographic data included femoral length, tibial length, total lower extremity length, mechanical axis deviation (MAD), lateral distal femoral angle (LDFA), and medial proximal tibial angle (MPTA). RESULTS: In total 12 adolescent patients with a mean age of 13.4 years (range, 12.3 to 14.4) and bone age of 13.4 years (11.5 to 14) at the time of surgery were included. At an average of 2.27-year follow-up (412 to 1058 d), there was no difference in the total growth of the operative and nonoperative limb (48.5 mm vs. 47 mm; P=0.36). In addition, the average increases in femoral length (23.4 mm) and tibial length (25.8 mm) were not statistically different between the operative and the nonoperative limb (P=0.12; P=0.75). There was no statistical difference in mechanical axis deviation, LDFA, or MPTA between preoperative and postoperative radiographs. Mean differences in operative and nonoperative coronal angular changes were all <1.5 degrees. CONCLUSIONS: With at least 2 years of growth remaining, transphyseal ACL reconstruction with anteromedial drilling did not significantly affect the physis or residual growth compared with the contralateral extremity. Although this technique may create a larger defect in the physis, standing radiographs demonstrate there is no change in limb length or angulation in growing adolescents approximately 2 years after surgery. LEVEL OF EVIDENCE: This is a case series; Level IV evidence.

Racial/Ethnic Disparities and Immunotherapeutic Advances in the Treatment of Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) remains one of the leading causes of death among many associated liver diseases. Various conventional strategies have been utilized for treatment, ranging from invasive surgeries and liver transplants to radiation therapy, but fail due to advanced disease progression, late screening/staging, and the various etiologies of HCC. This is especially evident within racially distinct populations, where incidence rates are higher and treatment outcomes are worse for racial/ethnic minorities than their Caucasian counterparts. However, with the rapid development of genetic engineering and molecular and synthetic biology, many novel strategies have presented promising results and have provided potential treatment options. In this review, we summarize past treatments, how they have shaped current treatments, and potential treatment strategies for HCC that may prove more effective in the future.

CS1 (SLAMF7) inhibits production of proinflammatory cytokines by activated monocytes.

OBJECTIVE AND DESIGN: CS1 (CRACC, CD319, SLAMF7) is a member of the Signaling Lymphocyte Activation Molecule family expressed on immune cells mediating host defense. CS1 is a self-ligand and has both activating and inhibitory functions in Natural Killer cells. However, the function of CS1 in human monocytes is currently unknown. The objective of this study was to evaluate the control of CS1 surface expression in activated monocytes and to assess the effect of CS1 triggering on proinflammatory cytokine production by monocytes. MATERIAL, METHODS AND TREATMENT: Human monocytes were isolated from PBMC of healthy volunteers by magnetic depletion method or FACS sorting. The monocytes were cultured with or without LPS (1 µg/ml) in the presence or absence of various pharmacological inhibitors to inhibit NF-кB and PI3K signaling pathways. The cells were stimulated with anti-CS1 antibody or isotype control. Total RNA was extracted and RT-PCR was performed using specific primers for CS1 and EAT-2. Cell supernatants were collected and cytokine levels (TNF-α and IL-12p70) were determined by sandwich ELISA. RESULTS: Our study revealed that adherent or LPS-activated monocytes express CS1, and CS1 induction is via NF-кB and PI3K pathways. Importantly, cross-linking CS1 resulted in reduced production of proinflammatory cytokines TNF-α and IL-12p70 by LPS-activated monocytes. CONCLUSIONS: Our study demonstrated that CS1 plays an inhibitory role in human monocytes to control proinflammatory immune responses.

Low-Intensity Pulsed Ultrasonography Plus Arthroscopic Drilling Does Not Improve Bone Healing More Than Arthroscopic Drilling Alone in Pediatric Patients With Stable Osteochondritis Dissecans of the Knee.

Purpose: To determine whether adjuvant use of bone stimulation would improve the rate of healing in the operative management of stable osteochondritis dissecans (OCD) of the knee in pediatric patients. Methods: This retrospective matched case-control study was performed at a single tertiary care pediatric hospital between January 2015 and September 2018. Patients who underwent antegrade drilling for stable femoral condyle OCD with greater than 2 years' follow-up were included. Preference was for all to receive postoperative bone stimulation; however, some were denied because of insurance coverage. This enabled us to create 2 matched groups of those who received postoperative bone stimulation and those who did not. Patients were matched on skeletal maturity, lesion location, sex, and age at surgery. The primary outcome measure was the rate of healing of the lesions determined by postoperative magnetic resonance imaging measurements at 3 months. Results: Fifty-five patients were identified who met the inclusion and exclusion criteria. Twenty patients from the bone stimulator group (BSTIM) were matched to 20 patients from the no bone stimulator group (NBSTIM). Mean age for BSTIM at surgery was 13.2 years ± 2.0 (range, 10.9-16.7) and for NBSTIM at surgery 12.9 years ± 2.0 (range, 9.3-17.3). At 2 years, 36 patients (90%) in both groups went on to clinical healing without further interventions. In BSTIM, there was a mean decrease of 0.9 (±1.8) mm in lesion on coronal width and 12 patients (63%) had overall improved healing; in NBSTIM there was a mean decrease of 0.8 (±3.6) mm in coronal width and 14 patients (78%) had improved healing. No statistical differences in the rate of healing were found between the 2 groups (P = .706). Conclusion: In antegrade drilling of stable knee OCD lesions in pediatric and adolescent patients, adjuvant bone stimulator use did not appear to improve radiographic or clinical healing. Level of evidence: Level III, retrospective case-control study.

Blocking PCNA interaction with NKp44 enhances primary natural killer cell-mediated lysis of triple-negative breast cancer cells.

Among the innate immune cells, natural killer cells (NK) serve its role in cytolytic targeting against infected and cancerous cells. NK function is regulated by an intricate balance of signals from interactions between activating and inhibitory NK receptors and ligands expressed on target cells. As an immune evasion strategy, cancer cells, particularly triple-negative breast cancer cells (TNBCs), express ligands that interact with NK receptors to inhibit NK cell cytolytic function. Our studies have revealed that Proliferating Cell Nuclear Antigen (PCNA), normally expressed in the nucleus with DNA replication and repair roles, was present on the cell surface of TNBC cell lines MDA-MB-231, -436, and -468. To elucidate the function of cell surface PCNA, we blocked PCNA on TNBCs with antibodies which both disrupted interaction with NKp44 and enhanced lysis by primary NK cells. Furthermore, a combinational antibody treatment of TNBCs with α-LLT1 and α-PCNA antibodies augments NK-mediated lysis. These results together suggest that cell surface PCNA on TNBCs enables evasion from cytolytic killing by NK cells. Blocking PCNA-NKp44 interaction with antibodies may potentially open an additional avenue in treatment of TNBCs.

Withaferin A: A Pleiotropic Anticancer Agent from the Indian Medicinal Plant Withania somnifera (L.) Dunal.

Cancer represents the second most deadly disease and one of the most important public health concerns worldwide. Surgery, chemotherapy, radiation therapy, and immune therapy are the major types of treatment strategies that have been implemented in cancer treatment. Unfortunately, these treatment options suffer from major limitations, such as drug-resistance and adverse effects, which may eventually result in disease recurrence. Many phytochemicals have been investigated for their antitumor efficacy in preclinical models and clinical studies to discover newer therapeutic agents with fewer adverse effects. Withaferin A, a natural bioactive molecule isolated from the Indian medicinal plant Withania somnifera (L.) Dunal, has been reported to impart anticancer activities against various cancer cell lines and preclinical cancer models by modulating the expression and activity of different oncogenic proteins. In this article, we have comprehensively discussed the biosynthesis of withaferin A as well as its antineoplastic activities and mode-of-action in in vitro and in vivo settings. We have also reviewed the effect of withaferin A on the expression of miRNAs, its combinational effect with other cytotoxic agents, withaferin A-based formulations, safety and toxicity profiles, and its clinical potential.

Natural Killer Cell-Mediated Immunotherapy for Leukemia.

Leukemia is a malignancy of the bone marrow and blood resulting from the abnormal differentiation of hematopoietic stem cells (HSCs). There are four main types of leukemia including acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), chronic myeloid leukemia (CML), and chronic lymphocytic leukemia (CLL). While chemotherapy and radiation have been conventional forms of treatment for leukemia, these therapies increase infection susceptibility, adverse side effects and immune cell inactivation. Immunotherapies are becoming promising treatment options for leukemia, with natural killer (NK) cell-mediated therapy providing a specific direction of interest. The role of NK cells is critical for cancer cell elimination as these immune cells are the first line of defense against cancer proliferation and are involved in both recognition and cytolysis of rapidly dividing and abnormal cell populations. NK cells possess various activating and inhibitory receptors, which regulate NK cell function, signaling either inhibition and continued surveillance, or activation and subsequent cytotoxic activity. In this review, we describe NK cells and NK cell receptors, functional impairment of NK cells in leukemia, NK cell immunotherapies currently under investigation, including monoclonal antibodies (mAbs), adoptive transfer, chimeric antigen receptor-NKs (CAR-NKs), bi-specific/tri-specific killer engagers (BiKEs/TriKEs) and future potential targets of NK cell-based immunotherapy for leukemia.

NK Cell Isolation and Cytotoxicity by Radioactive Chromium Release Assay and DELFIA-EuTDA Cytotoxicity Assay.

Cytotoxicity assays are important in vitro tools to measure the lysis of desired target cells via an effector immune cell of choice. Specific lysis of the target cells can be determined by labeling the target cells with a radioactive isotope or fluorescent molecule, co-incubating it with an effector cell, then measuring the release of the labeled molecule in the supernatant. Here, we describe and compare different cell cytotoxicity assays using a chromium-51 (51Cr) release and DELFIA EuTDA fluorescent assay using K562 as the target cells and peripheral blood mononuclear cell (PBMC) derived natural killer (NK) cells as the effector cells.

2B4+ CD8+ T cells play an inhibitory role against constrained HIV epitopes.

Cytotoxic T cells play a critical role in the control of HIV and the progression of infected individuals to AIDS. 2B4 (CD244) is a member of the SLAM family of receptors that regulate lymphocyte development and function. The expression of 2B4 on CD8+ T cells was shown to increase during AIDS disease progression. However, the functional role of 2B4+ CD8+ T cells against HIV infection is not known. Here, we have examined the functional role of 2B4+ CD8+ T cells during and after stimulation with HLA B14 or B27 restricted HIV epitopes. Interestingly, IFN-γ secretion and cytotoxic activity of 2B4+ CD8+ T cells stimulated with HIV peptides were significantly decreased when compared to influenza peptide stimulated 2B4+ CD8+ T cells. The expression of the signaling adaptor molecule SAP was downregulated in 2B4+ CD8+ T cells upon HIV peptide stimulation. These results suggest that 2B4+ CD8+ T cells play an inhibitory role against constrained HIV epitopes underlying the inability to control the virus during disease progression.

The Many Faces of Innate Immunity in SARS-CoV-2 Infection.

The innate immune system is important for initial antiviral response. SARS-CoV-2 can result in overactivity or suppression of the innate immune system. A dysregulated immune response is associated with poor outcomes; with patients having significant Neutrophil-to-Lymphocyte ratios (NLR) due to neutrophilia alongside lymphopenia. Elevated interleukin (IL)-6 and IL-8 leads to overactivity and is a prominent feature of severe COVID-19 patients. IL-6 can result in lymphopenia; where COVID-19 patients typically have significantly altered lymphocyte subsets. IL-8 attracts neutrophils; which may play a significant role in lung tissue damage with the formation of neutrophil extracellular traps leading to cytokine storm or acute respiratory distress syndrome. Several factors like pre-existing co-morbidities, genetic risks, viral pathogenicity, and therapeutic efficacy act as important modifiers of SARS-CoV-2 risks for disease through an interplay with innate host inflammatory responses. In this review, we discuss the role of the innate immune system at play with other important modifiers in SARS-CoV-2 infection.

Functional role of human NK cell receptor 2B4 (CD244) isoforms.

2B4 (CD244), a member of the signaling lymphocyte-activation molecule (SLAM/CD150), is expressed on all NK cells, a subpopulation of T cells, monocytes and basophils. Human NK cells express two isoforms of 2B4, h2B4-A and h2B4-B that differ in a small portion of the extracellular domain. In the present investigation, we have studied the functions of h2B4-A and h2B4-B. Our study demonstrated that these two isoforms differ in their binding affinity for CD48, which results in differential cytotoxic activity as well as intracellular calcium release by NK cells upon target cell recognition. Analysis of the predicted 3-D structure of the two isoforms showed conformational differences that could account for their differences in binding affinity to CD48. h2B4-A was able to mediate natural cytotoxicity against CD48-expressing K562 target cells and induce intracellular calcium release, whereas h2B4-B showed no effects. NK-92MI, U937, THP-1, KU812, primary monocytes, basophils and NK cells showed expression of both h2B4-A and h2B4-B whereas YT and IL-2-activated NK cells did not show any h2B4-B expression. Stimulation of NK cells through 2B4 resulted in decreased mRNA levels of both h2B4-A and h2B4-B indicating that down-regulation of 2B4 isoforms may be an important factor in controlling NK cell activation during immune responses.