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Coronavirus infection in humans is usually associated to respiratory tract illnesses, ranging in severity from mild to life-threatening respiratory failure. The aryl hydrocarbon receptor (AHR) was recently identified as a host factor for Zika and dengue viruses; AHR antagonists boost antiviral immunity, decrease viral titers and ameliorate Zika-induced pathology in vivo. Here we report that AHR is activated by infection with different coronaviruses, potentially impacting antiviral immunity and lung epithelial cells. Indeed, the analysis of single-cell RNA-seq from lung tissue detected increased expression of AHR and AHR transcriptional targets, suggesting AHR signaling activation in SARS-CoV-2-infected epithelial cells from COVID-19 patients. Moreover, we detected an association between AHR expression and viral load in SARS-CoV-2 infected patients. Finally, we found that the pharmacological inhibition of AHR suppressed the replication in vitro of one of the causative agents of the common cold, HCoV-229E, and the causative agent of the COVID-19 pandemic, SARS-CoV-2. Taken together, these findings suggest that AHR activation is a common strategy used by coronaviruses to evade antiviral immunity and promote viral replication, which may also contribute to lung pathology. Future studies should further evaluate the potential of AHR as a target for host-directed antiviral therapy., RNA viruses, such as Zika and Dengue, activate the aryl hydrocarbon receptor (AHR) signaling to restrict the antiviral response. Here, the authors report that coronaviruses also activate AHR signaling, potentially allowing them to escape the immune response, and show that AHR antagonism limits SARS-CoV-2 replication in vitro, suggesting that AHR could be targeted for antiviral therapy.

Background: Aging changes to the lower eyelids and midface include all but not only these changes: pseudoherniated orbital fat, tear trough deformity, lid laxity, and dermatochalasis. Surgical repair often aims at treating redundant skin or orbital fat malposition with a lower eyelid blepharoplasty. In manipulating the inferior orbital fat pads, a surgeon has many options including excision, repositioning, or augmentation with synthetic dermal filler, autologous fat grafts, or acellular dermal allografts [1]. The aim of this study is to find the best approach in preventing fat herniation reccurnce in lower lid blepharoplasty. Methods: The patients in study were classified into three groups depending on the used surgical technique, to test the most effective technique associated with minimal rate of lower fat pad recurrence after surgery. Two of these techniques include a muscular flap suspension from the orbicularis oculae muscle. Results: The two surgical techniques that include orbicularis oculi muscle suspension are associated with no recurrence of lower herniated fat pads after blepharoplasty. Conclusions: The suspension of orbicularis oculi muscle has an important role in enhancing the lower orbital septum and prevents the recurrence of the lower herniated fat pads.

Background: Resveratrol and Berberis integerrima (B. integerrima) are known to be natural antioxidants and regulators of human metabolism. However, the effects of resveratrol and B. integerrima on the ovarian morphology in polycystic ovary syndrome (PCOS) are not obvious. Objective: This study aimed to determine the effect of the hydroalcoholic extract of B. integerrima in combination with resveratrol on some biochemical parameters and ovarian morphology in the letrozole-induced PCOS rat. Materials and Methods: Seventy adult female Sprague-Dawley rats aged 10-12 weeks weighing 200 ± 20 gr were randomly divided into seven groups (n = 10/each). Group I): normal; Group II): vehicle; Group III): letrozole-induced PCOS 1 mg/kg letrozole orally, rats receiving 1 cc normal saline orally; Group IV): PCOS + receiving 150 mg/kg metformin orally; Group V): PCOS + receiving 20 mg/kg resveratrol orally; Group VI): PCOS + 3 gr/kg barberry orally; and Group VII): PCOS + receiving 3 gr/kg barberry and 20 mg/kg resveratrol orally. All animals were followed-up for 63 days. The biochemical parameters and histological assessments of ovaries were performed. Results: Resveratrol alone and/or in combination with B. integerrima treatment in rats led to a significant decrease in low-density lipoprotein, triglyceride, malondialdehyde , and tumor necrosis factor-alpha concentrations (p = 0.02). The groups IV, V, VI, and VII showed a decrease in insulin resistance and an increase in the superoxide dismutase, total antioxidant capacity, and high-density lipoprotein (p = 0.01). No significant difference was observed between the level of serum glucose in the treatment groups. Number of cystic follicles had a significant decrease in barberry, resveratrol, and their combination groups (p < 0.001). Conclusion: Resveratrol, B. integerrima, and their combination as natural products with fewer side effects might be effective as an alternative medicine in treatment of PCOS. Key words: Barberry, Resveratrol, Polycystic ovary syndrome, Ovary, Rat.

Background and Aim: The pathogenesis of schistosomiasis is mainly due to the host’s immune response against Schistosoma eggs trapped in tissues causing a granulomatous reaction. Recently, stem cell therapy has drawn much attention due to their potential for self-renewal and differentiation. The present study was designed to investigate the regenerative and antifibrotic potential of Human Umbilical Cord Blood derived Mesenchymal Stem Cells (HUCB-MSCs) in hepatic granuloma due to schistosomiasis mansoni in the experimental murine model. Material and Methods: Swiss Albino mice (n=70) were divided into 3 groups; Group A: 30 mice infected with S. mansoni cercariae and subjected to intrahepatic injection of HUCB-MSCs by a single dose of 1 million HUCB-MSCs at 12 weeks post infection (wpi), G (A1): 10 mice sacrificed at 16 (wpi), G (A2):10 mice sacrificed at 18 (wpi), G (A3): 10 mice sacrificed at 20 (wpi). Group B: 30 mice infected with S. mansoni cercariae (positive control), G (B1):10 mice sacrificed at 16 (wpi), G (B2):10 mice sacrificed at 18 (wpi), G (B3):10 mice sacrificed at 20 (wpi). Group C: 10 uninfected mice (negative control). Results: Treatment with HUCB-MSCs, showed an improvement in the histopathological picture of the liver with diminution in the size of granulomas and the overall fibrotic content. Hyperplasic changes in intrahepatic biliary radicals obviously decreased in test groups. Conclusion: The human umbilical cord blood-derived MSCs are promising and effective therapy for the treatment of diseased or damaged liver tissues.

Summary Although natural killer (NK) cells have become a promising immune effector cell for chimeric antigen receptor (CAR)-based therapy, generating human CAR-NK cells with high transgene efficiency has been challenging. In this protocol, we describe how to generate CAR-NK cells with transduction efficiencies >15% from healthy donor ex vivo expanded NK cells using third generation lentiviral vectors (LVs). We also show how to assess CAR-NK cell anti-tumor function in vitro using a flow cytometry-based killing assay. For complete details on the use and execution of this protocol, please refer to Portillo et al. (2021)., Graphical abstract, Highlights • PBMC isolation and expansion of human NK cells from healthy donors • Production of third generation lentiviral vectors for NK cell transduction • A protocol for the generation of CAR-modified NK cells from human expanded NK cells • A flow cytometry-based assay to assess CAR-NK cell cytotoxicity in vitro, Although natural killer (NK) cells have become a promising immune effector cell for chimeric antigen receptor (CAR)-based therapy, generating human CAR-NK cells with high transgene efficiency has been challenging. In this protocol, we describe how to generate CAR-NK cells with transduction efficiencies >15% from healthy donor ex vivo expanded NK cells using third generation lentiviral vectors (LVs). We also show how to assess CAR-NK cell anti-tumor function in vitro using a flow cytometry-based killing assay.

Cell membranes - primarily composed of lipids, sterols, and proteins - form a dynamic interface between living cells and their environment. They act as a mechanical barrier around the cell while selectively facilitating material transport, signal transduction, and various other functions necessary for the cell viability. The complex functionality of cell membranes and the hierarchical motions and responses they exhibit demand a thorough understanding of the origin of different membrane dynamics and how they are influenced by molecular additives and environmental cues. These dynamic modes include single-molecule diffusion, thermal fluctuations, and large-scale membrane deformations, to name a few. This review highlights advances in investigating structure-driven dynamics associated with model cell membranes, with a particular focus on insights gained from neutron scattering and spectroscopy experiments. We discuss the uniqueness of neutron contrast variation and its remarkable potential in probing selective membrane structure and dynamics on spatial and temporal scales over which key biological functions occur. We also present a summary of current and future opportunities in synergistic combinations of neutron scattering with molecular dynamics (MD) simulations to gain further understanding of the molecular mechanisms underlying complex membrane functions.

Introduction Although normal epithelial cells do not show human epidermal growth factor receptor-2 (HER2) gene amplification and should lack membrane staining by HER2 immunohistochemistry (IHC), HER2 staining in benign breast epithelium is occasionally encountered. The significance of this occurrence has not yet been adequately studied, and its associated American Society of Clinical Oncology/College of American Pathologists recommendations are vague. Our objective is to assess the correlation between HER2 IHC 3+ breast cancer cases with normal epithelium staining (NES) and their corresponding fluorescence in situ hybridization (FISH) results, and to suggest recommendations for interpretation. Materials and Methods A total of 154 breast cancer cases with HER2 IHC 3+ were reviewed. NES, along with other clinicopathologic characteristics, were recorded. NES was scored as present or absent. All study cases were sent for FISH testing. All cases, and particularly those that showed false positivity for IHC (positive IHC, negative FISH) were examined for NES. Results Of the 154 cases, 146 cases were FISH-positive (94.8%) and 2 failed FISH testing (1.3%). Conversely, 22% (34/154) of the cases showed NES for HER2. Of these 34 cases, 23 (67%) were FISH-amplified, 9 (26%) were FISH not amplified, and 2 failed FISH testing. Notably, all of the false-positive (FISH-negative) breast cancer cases showed some degree of positivity in normal breast epithelium. Conclusions Our findings, though descriptive, show a very strong association between NES and false-positive HER2 IHC. This confirms the need to carefully evaluate IHC-positive breast cancers for NES, and to have a low threshold for confirmatory testing by FISH.

Background: The uterus has a complex lymphatic drainage consisting of many groups that are in danger of endometrial cancer (EC) spread. Lymph node metastasis is a genuine pointer of poor prognosis requiring adjuvant therapy. Lymphatic mapping surgically can be done by blue dye labeling and perioperative algorithm. Aim: To assess the role of perioperative sentinel lymph nodes (SLN) mapping algorithm in reducing the necessity of pelvic lymphadenectomy. Methods: Fifty patients with early stage I EC underwent surgical staging. Transcervical injection of methylene blue was used, laparotomy was done and retroperitoneal spaces were opened. All lymph nodes were histopathologically examined and the negative SLN were ultrastaged. Results: Methylene blue dye injection resulted in blue lymph nodes in 20 (40%) cases, suspicious in 2 (4%) and negative in 28 (56%). Histopathological examination of sampled lymph node showed metastases in 18/20 (90%) of blue nodes, 1/2 (50%) of suspicious nodes and 0/28 (0%) of negative nodes. There were no false negative cases in this study and the detection rate was 89%. Conclusion: Perioperative algorithm accuracy and simplicity make it a reliable approach for lymph node assessment in early stage EC.

As highlighted by the COVID-19 global pandemic, elderly individuals comprise the majority of cases of severe viral infection outcomes and death. A combined inability to control viral replication and exacerbated inflammatory immune activation in elderly patients causes irreparable immune-mediated tissue pathology in response to infection. Key to these responses are type I, II, and III interferons (IFNs), which are involved in inducing an antiviral response, as well as controlling and suppressing inflammation and immunopathology. IFNs support monocyte/macrophage-stimulated immune responses that clear infection and promote their immunosuppressive functions that prevent excess inflammation and immune-mediated pathology. The timing and magnitude of IFN responses to infection are critical towards their immunoregulatory functions and ability to prevent immunopathology. Aging is associated with multiple defects in the ability of macrophages and dendritic cells to produce IFNs in response to viral infection, leading to a dysregulation of inflammatory immune responses. Understanding the implications of aging on IFN-regulated inflammation will give critical insights on how to treat and prevent severe infection in vulnerable individuals. In this review, we describe the causes of impaired IFN production in aging, and the evidence to suggest that these impairments impact the regulation of the innate and adaptive immune response to infection, thereby causing disease pathology.

Chronic kidney disease (CKD) and acute kidney injury (AKI) are common, heterogeneous, and morbid diseases. Mechanistic characterization of CKD and AKI in patients may facilitate a precision medicine approach to prevention, diagnosis, and treatment. The Kidney Precision Medicine Project aims to ethically and safely obtain kidney biopsies from participants with CKD or AKI, create a reference kidney atlas, and characterize disease subgroups to stratify patients based on molecular features of disease, clinical characteristics, and associated outcomes. An additional aim is to identify critical cells, pathways, and targets for novel therapies and preventive strategies. This project is a multicenter prospective cohort study of adults with CKD or AKI who undergo a protocol kidney biopsy for research purposes. This investigation focuses on kidney diseases that are most prevalent and therefore substantially burden the public health, including CKD attributed to diabetes or hypertension and AKI attributed to ischemic and toxic injuries. Reference kidney tissues (for example, living kidney donor biopsies) will also be evaluated. Traditional and digital pathology will be combined with transcriptomic, proteomic, and metabolomics analysis of the kidney tissue as well as deep clinical phenotyping for supervised and unsupervised subgroup analysis and systems biology analysis. Participants will be followed prospectively for ten years to ascertain clinical outcomes. Cell types, locations, and functions will be characterized in health and disease in an open, searchable, online kidney tissue atlas. All data from the Kidney Precision Medicine Project will be made readily available for broad use by scientists, clinicians, and patients.

HDGF-related protein 2 (HRP-2) is a member of the Hepatoma-Derived Growth Factor-related protein family that harbors the structured PWWP and Integrase Binding Domain, known to associate with methylated histone tails or cellular and viral proteins, respectively. Interestingly, HRP-2 is a paralog of Lens Epithelium Derived Growth Factor p75 (LEDGF/p75), which is essential for MLL-rearranged (MLL-r) leukemia but dispensable for hematopoiesis. Sequel to these findings, we investigated the role of HRP-2 in hematopoiesis and MLL-r leukemia. Protein interactions were investigated by co-immunoprecipitation and validated using recombinant proteins in NMR. A systemic knockout mouse model was used to study normal hematopoiesis and MLL-ENL transformation upon the different HRP-2 genotypes. The role of HRP-2 in MLL-r and other leukemic, human cell lines was evaluated by lentiviral-mediated miRNA targeting HRP-2. We demonstrate that MLL and HRP-2 interact through a conserved interface, although this interaction proved less dependent on menin than the MLL-LEDGF/p75 interaction. The systemic HRP-2 knockout mice only revealed an increase in neutrophils in the peripheral blood, whereas the depletion of HRP-2 in leukemic cell lines and transformed primary murine cells resulted in reduced colony formation independently of MLL-rearrangements. In contrast, primary murine HRP-2 knockout cells were efficiently transformed by the MLL-ENL fusion, indicating that HRP-2, unlike LEDGF/p75, is dispensable for the transformation of MLL-ENL leukemogenesis but important for leukemic cell survival. ispartof: CELLS vol:10 issue:1 ispartof: location:Switzerland status: published

Background: Hysteroscopy used to detect possible intrauterine causes ofinfertility. Misoprostol is a prostaglandin El analogue used in cervical ripeningbeforehysteroscopy to soften the cervix and decreasesthe force required for dilation. Objectives: Comparing the effectiveness of oral versus vaginal misoprostol administered 24 hours before hysteroscopy to facilitate the procedures in infertile patients. Patients and Methods:Our randomized control prospective study included 120 infertile women requiring hysteroscopy divided into three equal groups (Group A) received oral misoprostol with a dosage of 600 µg (200 µg / 8 hours), (Group B) receivedvaginal misoprostol with a dosage of 400 µg (200 µg /12 hours) and (group C) control group receivedoral placebo (one pill / 8 hours).Width of endocervical canal was assessed before the hysteroscope, the ease of entrance and the mean time needed for cervical dilation. Results: Statistically substantial differences among the oral and vaginal groups (p < 0,009) and among the vaginal and control groups and the oral and control groups (p < 0,001 for both).There was a substantial difference in the ease of cervical entry among the vaginal and control groups and the oral and control groups (p < 0,001 for both), but not among the vaginal and oral groups (p < 0,998). The variation in time was not substantial between both the oral and vaginal groups. Conclusion: Misoprostol is a good cervical ripening agent. Themove from the vaginal to the oral path, which more compliance by the patients, could applied without incurring a risk to clinical effectiveness.

Summary NK cells are central to anti-tumor immunity and recently showed efficacy for treating hematologic malignancies. However, their dysfunction in the hostile tumor microenvironment remains a pivotal barrier for cancer immunotherapies against solid tumors. Using cancer patient samples and proteomics, we found that human NK cell dysfunction in the tumor microenvironment is due to suppression of glucose metabolism via lipid peroxidation-associated oxidative stress. Activation of the Nrf2 antioxidant pathway restored NK cell metabolism and function and resulted in greater anti-tumor activity in vivo. Strikingly, expanded NK cells reprogrammed with complete metabolic substrate flexibility not only sustained metabolic fitness but paradoxically augmented their tumor killing in the tumor microenvironment and in response to nutrient deprivation. Our results uncover that metabolic flexibility enables a cytotoxic immune cell to exploit the metabolic hostility of tumors for their advantage, addressing a critical hurdle for cancer immunotherapy.

Background: Few studies including limited number of patients assessed the rheumatologic effects of hemodialysis (HD) on joints using ultrasonography. Joint ultrasound has been emerged as a cheap non-invasive tool for assessment of joint pain among HD patients. This was the aim of our study to make use of such tool in such life quality threatening complaint. Objective: to determine the role of ultrasound in evaluation of joint pain and its causes among patients on regular HD. Patients and Methods: One hundred and four patients with end stage renal disease (ESRD) who were regular on HD three sessions per week four hours per session were subjected to history taking, complete physical examination stressing on musculoskeletal examination and ultrasonography of painful joints by an ultrasonography expert. Results: Dialysis related arthropathy (DRA) was not the only cause of joint pain among HD patients but there were diverse causes in different joints. As regard affected joints, knee was the most affected one then came wrist, shoulder, ankle and elbow respectively. As regard causes of joint pain, DRA was the commonest one then came osteoarthritis, non-specific ultrasonographic findings and few cases showed normal ultrasonographic studies. Conclusion: This study confirmed that joint pain in HD patients has diverse causes not DRA by necessity but other causes must be considered as well as multifactorial etiologies.

Introduction This study investigated the impact of different protocols on radiation dose and image quality for obese patients undergoing abdominal CT examinations. Methods Five abdominal/pelvis CT protocols employed across three scanners from a single manufacturer in a single centre used a variety of parameters (kV: 100/120, reference mAs: 150/190/218/250/300, image reconstruction: filtered back projection (FBP)/iterative (IR)). The routine protocol employed 300 reference mAs and 120 kV. Data sets resulting from obese patient examinations ( n = 42) were assessed for image quality using visual grading analysis by three experienced radiologists. Objective assessment (noise, signal/contrast-noise ratios) and radiation dose was compared to determine optimal protocols for prospective testing on a further sample of patients ( n = 47) for scanners using FBP and IR techniques. Results Compared to the routine protocol, mean radiation dose was reduced by 60% when using 100 kV and SAFIRE technique strength 3 ( p = 0.001). Reduction of up to 30% in radiation dose was noted for the FBP protocol: 120 kV and 190 reference mAs ( p = 0.008). Subjective and objective image quality for both protocols were comparable to that of the routine protocol ( p > 0.05). An overall improvement in image quality with increasing strength of SAFIRE was noted. Upon clinical implementation of the optimal dose protocols, local radiology consensus deemed image quality to be acceptable for the participating obese patient cohort. Conclusion Radiation dose for obese patients can be optimised whilst maintaining image quality. Where iterative reconstruction is available relatively low kV and quality reference mAs are also viable for imaging obese patients at 30–60% lower radiation doses.

Ovarian cancer (OC) is the leading cause of gynecological cancer-related death in North America. Most ovarian cancer patients (OCPs) experience disease recurrence after first-line surgery and chemotherapy; thus, there is a need for novel second-line treatments to improve the prognosis of OC. Although peripheral blood-derived NK cells are known for their ability to spontaneously lyse tumour cells without prior sensitization, ascites-derived NK cells (ascites-NK cells) isolated from OCPs exhibit inhibitory phenotypes, impaired cytotoxicity and may play a pro-tumourigenic role in cancer progression. Therefore, it is of interest to improve the cytotoxic effector function of impaired OCP ascites-NK cells at the tumour environment. We investigated the efficacy of using an artificial APC-based ex vivo expansion technique to generate cytotoxic, expanded NK cells from previously impaired OCP ascites-NK cells, for use in an autologous model of NK cell immunotherapy. We are the first to obtain a log-scale expansion of OCP ascites-NK cells that upregulate the surface expression of activating receptors NKG2D, NKp30, NKp44, produce robust amounts of anti-tumour cytokines in the presence of OC cells and mediate direct tumour cytotoxicity against ascites-derived, primary OC cells obtained from autologous patients. Our findings demonstrate that it is possible to generate cytotoxic OCP ascites-NK cells from previously impaired OCP ascites-NK cells, which presents a promising immunotherapeutic target for the second-line treatment of OC. Future work should focus on evaluating the in vivo efficacy of autologous NK cell immunotherapy through the intraperitoneal delivery of NK cell expansion factors to a preclinical xenograft mouse model of human OC.

High intensity interval training (HIIT) boosts natural killer (NK) cell number and activity in normal weight breast cancer patients; however, whether this occurs in obese individuals is not well established. The goal of this study was to determine whether HIIT effectively boosts NK cells as a therapeutic strategy against breast cancer in an obese mouse model and in overweight/obese women. Diet induced female C57Bl/6 obese mice were assigned to undergo HIIT for four weeks or remain sedentary. Female participants were subjected to a six weeks HIIT protocol. HIIT mice acclimatized to treadmill running were subsequently injected with 5 × 105 polyoma middle T (MT) breast cancer cells intravenously. NK cell number and activation were monitored using flow cytometry, and tumor burden or lipid content evaluated from histological lung and liver tissues, respectively. In both mice and humans, circulating NK cell number and activation (CD3-NK1.1+CD27+ and CD3-CD56+, respectively) markedly increased immediately after HIIT. HIIT obese mice had reduced lung tumor burden compared to controls following MT challenge, and had diminished hepatic lipid deposition despite minimal body weight loss. Our findings demonstrate that HIIT can benefit obese individuals by enhancing NK cell number and activity, reducing tumor burden, and enhancing metabolic health.

Background The translation of preclinically promising novel tuberculosis vaccines to ultimate human applications has been challenged by the lack of animal models with an immune system equivalent to the human immune system in its genetic diversity and level of susceptibility to tuberculosis. Methods We have developed a humanized mice (Hu-mice) tuberculosis model system to investigate the clinical relevance of a novel virus-vectored (VV) tuberculosis vaccine administered via respiratory mucosal or parenteral route. Results We find that VV vaccine activates T cells in Hu-mice as it does in human vaccinees. The respiratory mucosal route for delivery of VV vaccine in Hu-mice, but not the parenteral route, significantly reduces the humanlike lung tuberculosis outcomes in a human T-cell-dependent manner. Conclusions Our results suggest that the Hu-mouse can be used to predict the protective efficacy of novel tuberculosis vaccines/strategies before they proceed to large, expensive human trials. This new vaccine testing system will facilitate the global pace of clinical tuberculosis vaccine development.

Background: Intraparenchymal thyroid Doppler measurements might be considered a useful index of the thyroid status as well as micro-circulation elsewhere in the body among sickle cell disea...

NK cells are a key antiviral component of the innate immune response to HSV-2, particularly through their production of IFN-γ. It is still commonly thought that type I IFN activates NK cell function; however, rather than requiring the type I IFN receptor themselves, we have previously found that type I IFN activates NK cells through an indirect mechanism involving inflammatory monocytes and IL-18. Here, we further show that direct action of type I IFN on NK cells, rather than inducing IFN-γ, negatively regulates its production during HSV-2 infection and cytokine stimulation. During infection, IFN-γ is rapidly induced from NK cells at day 2 post-infection and then immediately downregulated at day 3 post-infection. We found that this downregulation of IFN-γ release was not due to a loss of NK cells at day 3 post-infection, but negatively regulated through IFN signaling on NK cells. Absence of IFNAR on NK cells led to a significantly increased level of IFN-γ compared to WT NK cells after HSV-2 infection in vitro. Further, priming of NK cells with type I IFN was able to suppress cytokine-induced IFN-γ production from both human and mouse NK cells. We found that this immunosuppression was not mediated by IL-10. Rather, we found that type I IFN induced a significant increase in Axl expression on human NK cells. Overall, our data suggests that type I IFN negatively regulates NK cell IFN-γ production through a direct mechanism in vitro and during HSV-2 infection.

NK cells are capable of an array of functions that range widely from their classic anti-tumor and anti-viral cytotoxic effector functions, to their critical regulatory roles in controlling inflammatory immune responses and promoting tissue growth. However, the mechanisms that polarize NK cells to these distinct and opposing functions are incompletely understood. NK cell functional subsets are primarily identified and studied based on phenotype, which has served as an accessible means for profiling NK cells and does offer information on NK cell activation state. However, inconsistencies have emerged in using classic phenotypes to inform function, which raise the questions: Can phenotype in fact define NK cell functional fate? What factors do profile and drive NK cell fate? In other immune cells, cell metabolism has been shown to critically determine subset polarization. There is a growing body of evidence that cell metabolism is integral to NK cell effector functions. Glucose-driven glycolysis and oxidative metabolism have been shown to drive classic NK cell anti-tumor and anti-viral effector functions. Recent studies have uncovered a critical role for metabolism in NK cell development, education, and memory generation. In this review, we will draw on the evidence to date to investigate the relationship between NK cell phenotype, metabolism, and functional fate. We explore a paradigm in which the differential activity of metabolic pathways within NK cells produce distinct metabolic fingerprints that comprehensively distinguish and drive the range of NK cell functional abilities. We will discuss future areas of study that are needed to develop and test this paradigm and suggest strategies to efficiently profile NK cells based on metabolism. Given the emerging role of metabolism in driving NK cell fates, profiling and modulating NK cell metabolism holds profound therapeutic potential to tune inflammatory and regulatory NK cell responses to treat disease.

Background Few studies including limited number of patients assessed the rheumatologic effects of hemodialysis (HD) on joints using ultrasonography. Joint ultrasound has been emerged as a cheap noninvasive tool for assessment of joint pain among HD patients. This was the aim of our study to make use of such tool in such life quality threatening complaints Objectives to determine the role of ultrasound in evaluation of joint pain and its causes among patients on regular HD Methods One hundred and four patients with end stage renal disease (ESRD) who were regular on HD three sessions per week four hours per session were subjected to history taking, complete physical examination stressing on musculoskeletal examination and ultrasonography of painful joints. Results Dialysis related arthropathy (DRA) was not the only cause of joint pain among HD patients but there were diverse causes in different joints. As regard affected joints, knee was the most affected one then came wrist, shoulder, ankle and elbow respectively. As regard causes of joint pain, DRA was the commonest one then came osteoarthritis, Nonspecific ultrasonographic findings and few cases showed normal ultrasonographic studies. Conclusion This study confirmed that joint pain in HD patients has diverse causes not DRA by necessity but other causes must be considered as well as multifactorial etiologies. Disclosure of Interests None declared

Objectives The aim of this paper is to evaluate contrast media (CM) bolus geometry and opacification patterns in the coronary arteries with particular focus on patient, scanner and safety considerations during coronary computed tomography angiography (CCTA). Key findings The rapid evolution of computed tomography (CT) technology has seen this imaging modality challenge conventional coronary angiography in the evaluation of coronary artery disease. Increases in spatial and temporal resolutions have enabled CCTA to become the modality of choice when evaluating the coronary vascular tree as an alternative in the diagnostic algorithm for acute chest pain. However, these new technologic improvements in scanner technology have imposed new challenges for the optimisation of CM delivery and image acquisition strategies. Conclusion Understanding basic CM-imaging principles is essential for designing optimal injection protocols according to each specific clinical scenario, independently of scanner technology. Implications for practice With rapid advances in CT scanner technology including faster scan acquisitions, the risk of poor opacification of coronary vasculature increases significantly. Therefore, awareness of CM delivery protocols is paramount to consistently provide optimal image quality at a low radiation dose.

5018 Background: The optimal management of patients (pts) with complete response (CR) after first-line chemotherapy remains unsettled with guidelines recommending either surveillance or retroperitoneal LN dissection (RPLND). We present long-term outcomes from a large dataset of pts managed with surveillance after achieving CR to first-line chemotherapy. Methods: The prospectively maintained Indiana University testicular cancer database was queried for pts with metastatic NSGCT treated between 1990-2017 who achieved a CR after first-line chemotherapy. CR was defined as normalization of tumor markers (AFP+hCG) and no residual mass > 1cm. Kaplan-Meier methods were used to analyze progression-free survival (PFS) and overall survival (OS). Results: 388 pts met eligibility and were included in this analysis. Median age at diagnosis was 28.4 (range, 13-61.5). Primary site was testis in 385 pts (99%). Primary tumor predominant histology was embryonal ca (241), mixed (61), seminoma (31), yolk sac tumor (20), choriocarcinoma (10), and teratoma (14). 126 pts (32.5%) had teratoma in the primary tumor. Metastasis sites were retroperitoneum (295), mediastinal LN (15), pulmonary (149), liver (15), bone (7), and brain (6). IGCCCG risk was good in 325, intermediate in 25, and poor in 32 pts. Pre-chemotherapy retroperitoneal LN size was available in 232 pts: < 3cm in 170 and ≥3cm in 62. Median pre-chemo AFP was 10.7 (1-31,000) and hCG was 16.5 (0-595,930). First-line chemo was BEPx3 in 274, BEPx4 in 30, other regimens in 82 pts. With a median follow-up of 3.9 yrs, 34 pts (8.8%) progressed. At most recent follow-up, 363 (93.6%) pts were alive with no evidence of disease and 10 pts (2.6%) died of their disease. The estimated 2-yr PFS was 90.1% (95% CI: 86.2-93%) and 2-yr OS was 97.8% (95% CI: 95.2-99%). The estimated 2-yr PFS by IGCCCG risk category was 90.4% for good vs 90.4% for intermediate vs 86.5% for poor risk (p = 0.23), and the estimated 2-yr OS was 98.6% for good vs 95.5% for intermediate vs 92.9% for poor risk disease respectively (p = 0.002). For the 34 pts who progressed on surveillance, 16 (4%) progressed in the retroperitoneum only. 3 pts had malignant transformation of teratoma to PNET, adenocarcinoma, or other elements. 11 of progressed pts were treated with surgery, 12 were treated with salvage chemo, and 11 were treated with surgery+chemo. At most recent follow up, 21 of progressed pts had NED, 10 had died of disease, and 3 were lost to follow up. Conclusions: Pts with metastatic NSGCT who achieve CR after first-line chemotherapy can be safely observed with surveillance. Most pts who relapse can be salvaged with surgery and/or chemotherapy.

A subset of innate lymphoid cells (ILCs), which produce IL-22 and function in protection against microbial infection, is generated via a local process in gut that is conditioned by commensal bacteria. We investigated the possibility that modulation of ILCs may contribute to the protective effect of certain bacteria in relation to pathogen infection. We fed BALB/c mice with L. rhamnosus JB-1 and characterized Peyer's patches NKp46+ cells. Additionally, the effect of feeding L. rhamnosus JB-1 on S. enterica serovar Typhimurium infection was determined. L. rhamnosus JB-1-feeding resulted in lower bacterial loads of Salmonella Typhimurium in the Peyer's patches, mesenteric lymph nodes and colon; and was associated with high levels of IL-22, RORγt, and CD127 expression by NKp46+ cells indicating an increase in ILC3s. Our findings indicate that Lactobacillus feeding regulates the innate lymphoid cell population that may contribute to host defensive function during Salmonella infection.

Prostate cancer (PCa) was estimated to have the second highest global incidence rate for male non-skin tumors and is the fifth most deadly in men thus mandating the need for novel treatment options. MG1-Maraba is a potent and versatile oncolytic virus capable of lethally infecting a variety of prostatic tumor cell lines alongside primary PCa biopsies and exerts direct oncolytic effects against large TRAMP-C2 tumors in vivo. An oncolytic immunotherapeutic strategy utilizing a priming vaccine and intravenously administered MG1-Maraba both expressing the human six-transmembrane antigen of the prostate (STEAP) protein generated specific CD8+ T-cell responses against multiple STEAP epitopes and resulted in functional breach of tolerance. Treatment of mice with bulky TRAMP-C2 tumors using oncolytic STEAP immunotherapy induced an overt delay in tumor progression, marked intratumoral lymphocytic infiltration with an active transcriptional profile and up-regulation of MHC class I. The preclinical data generated here offers clear rationale for clinically evaluating this approach for men with advanced PCa.

Metabolic flux can dictate cell fate, including immune cell effector and regulatory function. The metabolic regulation of cell function is well characterized with respect to effector, memory, and regulatory T cells. This knowledge may allow for manipulation of T cell metabolic pathways that set the stage for more effective T cell therapy. Natural Killer (NK) and T-lymphocytes have complementary roles in the defense against pathogens. However, studies of NK cell metabolism are only beginning to emerge and there is comparatively little knowledge on the metabolic regulation of NK-cell activation and effector function. Given their common lymphoid lineage, effector functions and cellular memory potential our current knowledge on T cell metabolism could inform investigation of metabolic reprogramming in NK cells. In this review, we compare the current knowledge of metabolic regulation in T cell and NK cell development, activation, effector and memory function. Commonalties in glucose transport, hypoxia-inducible factors and mTOR highlight metabolic control points in both cells types. Contrasting the glycolytic and oxidative nodes of metabolic regulation in T cells versus NK cells may provide insight into the contribution of specific immune responses to disease and promote the development of immunotherapeutic approaches targeting both innate and adaptive immune responses.

With over 600,000 units of umbilical cord blood (CB) stored on a global scale, it is important to elucidate the therapeutic abilities of this cryopreserved reservoir. In the advancing field of natural killer (NK) cell cancer immunotherapy, CB has proven to be a promising and noninvasive source of therapeutic NK cells. Although studies have proven the clinical efficacy of using long-term cryopreserved CB in the context of hematopoietic stem cell transplantations, little is known about its use for the ex vivo expansion of effector immune cells. Therefore, our group sought to derive ex vivo-expanded NK cells from long-term cryopreserved CB, using an artificial antigen presenting cell-mediated expansion technique. We compared the expansion potential and antitumor effector function of CB-derived NK (CB-NK) cells expanded from fresh (n=4), short-term cryopreserved (

Cell-based therapy is emerging as a promising therapeutic approach for a wide range of liver diseases. This study aimed to investigate the regenerative and antifibrotic therapeutic potential of bone marrow-derived mesenchymal stem cells (BM-MSCs) in an early and late experimental hepatic schistosomiasis model. BM-MSCs were isolated from 6-wk-old BALB/c donor male mice, then grown and propagated in culture until cell count was 5-8 × 10(6)/ml. MSCs were then separated and injected into Schistosoma mansoni -infected female BALB/c mice on their 6, 10, 14, and 18 wk post-infection. Mice were sacrificed on the fourth and eighth week after BM-MSCs transplantation in each group. Homing of BM-MSCs was confirmed by PCR detection of male Y-chromosome gene (sry) in the liver tissue of the recipient female mice. The regenerative and antifibrotic potential of BM-MSCs was assessed by histopathological examination, morphometric analysis, electron microscopy, and liver function tests. Schistosoma-infected mice, which were treated with BM-MSCs, showed a decrease in the granuloma size, percentage and density of the fibrotic area, formation of new hepatocytes, and improvement of the liver function tests. Immunohistochemical examination of alpha-smooth muscle actin revealed a significant decrease in the immunoreactive hepatic stellate cells in mice treated with MSCs. Early granulomas (acute infection) showed better response to MSC injection than did later granulomas (chronic infection). Dosing and timing of MSCs transplantation should undergo more investigations in long-term experiments before application to the clinical field. This study is the first to assess and compare the effect of MSCs treatment on early and late granulomas.

Plasmonic photothermal therapy (PPTT) was introduced as a promising treatment of cancer. This work was conducted to evaluate the cytotoxic effect of intratumoral (IT) injection of 75μg gold nanorods (GNRs)/kg of body weight followed by direct exposure to 2 w/cm 2 near infra-red laser light for 10min on ablation of mammary tumor in 10 dogs and 6 cats. Complete blood count (CBC), liver and kidney function were checked before the start of treatment and one month after injection of GNRs. Results showed that 62.5% (10/16), 25% (4/16) and 12.5% (2/16) of treated animals showed complete remission, partial remission and no response, respectively. Tumor was relapsed in 4 cases of initially responding animals (25%). Overall survival rate was extended to 315.5±20.5days. GNRs have no toxic effect on blood profile, liver or kidney functions. In conclusion, GNRs can be safely used for treatment of mammary tumors in dogs and cats.

Approximately 40% of HIV-1 infections occur in the female genital tract (FGT), primarily through heterosexual transmission. FGT factors determining outcome of HIV-1 exposure are incompletely understood, limiting prevention strategies. Here, humanized NOD-Rag1−/− γc−/− mice differentially reconstituted with human CD34+ -enriched hematopoietic stem cells (Hu-mice), were used to assess target cell frequency and viral inoculation dose as determinants of HIV-1 infection following intravaginal (IVAG) challenge. Results revealed a significant correlation between HIV-1 susceptibility and hCD45+ target cells in the blood, which correlated with presence of target cells in the FGT, in the absence of local inflammation. HIV-1 plasma load was associated with viral dose at inoculation and frequency of target cells. Events following IVAG HIV-1 infection; viral dissemination and CD4 depletion, were not affected by these parameters. Following IVAG inoculation, HIV-1 titres peaked, then declined in vaginal lavage while plasma showed a reciprocal pattern. The greatest frequency of HIV-1-infected (p24+) cells were found one week post-infection in the FGT versus blood and spleen, suggesting local viral amplification. Five weeks post-infection, HIV-1 disseminated into systemic tissues, in a dose-dependent manner, followed by depletion of hCD45+ CD3+ CD4+ cells. Results indicate target cell frequency in the Hu-mouse FGT is a key determinant of HIV-1 infection, which might provide a useful target for prophylaxis in women.

This retrospective cohort study of 2051 consecutive fresh non-donor intracytoplasmic sperm injection (ICSI) cycles investigated whether time from oocyte retrieval to denudation, precisely measured and recorded by an operator-independent automated radiofrequency-based system, affected cycle outcome. ICSI cycles were divided into two groups: group I (denudation within

Thyroid hormone receptor (THR) α and THRβ mediate the genomic action of thyroid hormones (THs) that affect bovine embryo development. However, little is known about THRs in the preimplantation embryo. The aim of the present study was to investigate the importance of THRs in in vitro preimplantation bovine embryos. THR transcripts and protein levels were detected in developing preimplantation embryos up to the blastocyst stage. Embryonic transcription of THRs was inhibited by α-amanitin supplementation, and both maternal and embryonic transcription were knocked down by short interference (si) RNA microinjection. In the control group, mRNA and protein levels of THRs increased after fertilisation. In contrast, in both the transcription inhibition and knockdown groups there were significant (P

Intestinal T helper type 2 (Th2) immunity in food allergy results in IgG1 and IgE production, and antigen re-exposure elicits responses such as anaphylaxis and eosinophilic inflammation. Although interleukin-4 (IL-4) is critically required for allergic sensitization, the source and control of IL-4 during the initiation of Th2 immunity in vivo remains unclear. Non-intestinal and non-food allergy systems have suggested that natural killer-like T (NKT) or γδ T-cell innate lymphocytes can supply the IL-4 required to induce Th2 polarization. Group 2 innate lymphoid cells (ILCs) are a novel IL-4-competent population, but their contribution to initiating adaptive Th2 immunity is unclear. There are also reports of IL-4-independent Th2 responses. Here, we show that IL-4-dependent peanut allergic Th2 responses are completely intact in NKT-deficient, γδ T-deficient or ILC-deficient mice, including antigen-specific IgG1/IgE production, anaphylaxis, and cytokine production. Instead, IL-4 solely from CD4(+) Th cells induces full Th2 immunity. Further, CD4(+) Th cell production of IL-4 in vivo is dependent on OX40L, a costimulatory molecule on dendritic cells (DCs) required for intestinal allergic priming. However, both Th2 cells and ILCs orchestrated IL-13-dependent eosinophilic inflammation. Thus, intestinal Th2 priming is initiated by an autocrine/paracrine acting CD4(+) Th cell-intrinsic IL-4 program that is controlled by DC OX40L, and not by NKT, γδ T, or ILC cells.

Interaction of mycobacteria with the host leads to retarded expression of T helper cell type 1 (Th1) immunity in the lung. However, the immune mechanisms remain poorly understood. Using in vivo and in vitro models of Mycobacterium tuberculosis (M. tb) infection, we find the immunoadaptor DAP12 (DNAX-activating protein of 12 kDa) in antigen-presenting cells (APCs) to be critically involved in this process. Upon infection of APCs, DAP12 is required for IRAK-M (interleukin-1 receptor-associated kinase M) expression, which in turn induces interleukin-10 (IL-10) and an immune-suppressed phenotype of APCs, thus leading to suppressed Th1 cell activation. Lack of DAP12 reduces APC IL-10 production and increases their Th1 cell-activating capability, resulting in expedited Th1 responses and enhanced protection. On the other hand, adoptively transferred DAP12-competent APCs suppress Th1 cell activation within DAP12-deficient hosts, and blockade of IL-10 aborts the ability of DAP12-competent APCs to suppress Th1 activation. Our study identifies the DAP12/IRAK-M/IL-10 to be a novel molecular pathway in APCs exploited by mycobacterial pathogens, allowing infection a foothold in the lung.

The immune mechanisms underlying delayed induction of Th1-type immunity in the lungs following pulmonary mycobacterial infection remain poorly understood. We have herein investigated the underlying immune mechanisms for such delayed responses and whether a selected innate immune-modulating strategy can accelerate Th1-type responses. We have found that, in the early stage of pulmonary infection with attenuated Mycobacterium tuberculosis (M.tb H37Ra), the levels of infection in the lung continue to increase logarithmically until days 14 and 21 postinfection in C57BL/6 mice. The activation of innate immune responses, particularly DCs, in the lung is delayed. This results in a delay in the subsequent downstream immune responses including the migration of antigen-bearing DCs to the draining lymph node (dLN), the Th1-cell priming in dLN, and the recruitment of Th1 cells to the lung. However, single lung mucosal exposure to the TLR agonist FimH postinfection is able to accelerate protective Th1-type immunity via facilitating DC migration to the lung and draining lymph nodes, enhancing DC antigen presentation and Th1-cell priming. These findings hold implications for the development of immunotherapeutic and vaccination strategies and suggest that enhancement of early innate immune activation is a viable option for improving Th1-type immunity against pulmonary mycobacterial diseases.

[This corrects the article DOI: 10.1371/journal.pgen.1007785.].

Background: Breast cancer therapy, which includes surgery, radiotherapy, chemotherapy and/or systemic therapy can have a profound impact on reproductive functions, leading to loss of fertility. To our knowledge, this is the first report on cancer photothermal therapy using gold nanorods on the mammary gland treatment of a cat, and the safe effect of the treatment on the reproductive function after tumor remission. Case presentation: A seven years old Shirazi cat (Toatoa) was evaluated because of a 2-week history of progressive dyspnea, signs of depression, and loss of appetite. The cat has a large tumor mass at the left caudal mammary gland. The diameter of the tumor mass was measured using caliper with the dimensions of 14 × 12 × 10.5 cm for length, width and depth, respectively. This was confirmed with ultrasonography. Biopsy samples were taken and fixed in 10% formalin for histopathological investigation, and it was diagnosed as mammary gland adenocarcinoma Grade II. Toatoa was injected intratumoral (IT) with 75 μg gold nanorods (GNRs)/kg body weight followed by exposure to 808 nm laser light for 10 min. GNRs were injected twice, with 15 days apart. After 15 days from the first GNRs injection, there was 60% ablation of the tumor size, while, after 17 days from the second GNRs dose, there was a complete tumor remission. Ultrasound scanning revealed complete ablation of the tumor mass. Complete blood picture (CBC), liver and kidney function analyses showed no changes in any of the tested parameters and indicated that GNRs photothermal treatment is safe and have no immediate toxic effects. After complete remission of the mammary gland tumor, the cat restores all the biological activities including the reproductive function. After 2 months from complete tumor remission, the cat was pregnant after mating with a fertile male, and 62 days later she delivered 3 kittens of normal morphology and growth rate. Breast feeding was found to be normal from all the nipples including the previously affected nipple. Conclusion: Photothermal therapy with gold nanorods can be used for the treatment of mammary gland tumor with apparently no impairments of reproductive functions in cats.

The priming of natural killer (NK) cells by type I interferon (IFN) is necessary for protection against primary and secondary viral infections. However, the pathway by which type I IFN activates NK cells to elicit antiviral responses is controversial. There is evidence to suggest that type I IFN priming of NK cells occurs through both direct and indirect pathways. As with many innate mechanisms, type I IFN and NK cells also orchestrate the adaptive immune response and thus aid in protection against secondary infections. Type I IFN can shape CD4(+) T cell, B cell and humoral memory formation. In addition, long-lived NK cells can perform specific and enhanced memory-like protection in secondary infections. This review outlines the different mechanisms underlying type I IFN regulation of NK cells and how type I IFN and NK cells can be used as a therapeutic target in vaccinations.

Lymphangioma circumscriptum (LC) is a form of lymphangioma characterized by benign dilation of lymphatic channels, which affects the skin and subcutaneous tissues. The most common sites are mucosa of mouth, tongue, groin, axilla, trunk and proximal region of extremities. Vulva is a rare site of LC. These abnormal lymphatic malformations do not communicate to the normal lymphatics. A female patient, aged 17 years, presented with papule-like condyloma of the labia majora. After simple vulvectomy was performed, symptoms improved. Histology revealed lymphangioma circumscriptum of the vulva. LC is a rare benign pathology that should be kept in mind in any vulval condylomatous lesion. Till date, only a few cases of LC of vulva have been reported in the literature. The treatment of primary vulvar lymphangioma circumscriptum is mainly surgery. Depending on the age of the patient and the extent of lesion, surgical treatment might be in the form of the local excision, labiectomy, or vulvectomy.

So far, the characteristics of a good quality egg have been elusive, similar to the nature of the physiological, cellular, and molecular cues leading to its production both in vivo and in vitro. Current understanding highlights a strong and complex interdependence between the follicular cells and the gamete. Secreted factors induce cellular responses in the follicular cells, and direct exchange of small molecules from the cumulus cells to the oocyte through gap junctions controls meiotic arrest. Studying the interconnection between the cumulus cells and the oocyte, we previously demonstrated that the somatic cells also contribute transcripts to the gamete. Here, we show that these transcripts can be visualized moving down the transzonal projections (TZPs) to the oocyte, and that a time course analysis revealed progressive RNA accumulation in the TZPs, indicating that RNA transfer occurs before the initiation of meiosis resumption under a timetable fitting with the acquisition of developmental competence. A comparison of the identity of the nascent transcripts trafficking in the TZPs, with those in the oocyte increasing in abundance during maturation, and that are present on the oocyte's polyribosomes, revealed transcripts common to all three fractions, suggesting the use of transferred transcripts for translation. Furthermore, the removal of potential RNA trafficking by stripping the cumulus cells caused a significant reduction in maturation rates, indicating the need for the cumulus cell RNA transfer to the oocyte. These results offer a new perspective to the determinants of oocyte quality and female fertility, as well as provide insight that may eventually be used to improve in vitro maturation conditions.

Uterine natural killer (NK) cells are pivotal for successful mammalian reproduction. However, insights on functionally distinct subpopulations of uterine NK cells are largely elusive. Furthermore, translation of findings from murine into human pregnancy has been overshadowed by the limited number of mutual phenotypic NK cell markers. We here provide evidence that a subset of murine mature NK (mNK) cells present at the feto-maternal interface, identified as CD27(low)DX5(+)CD3(neg), is pivotal in maintaining pregnancy. This mNK subset has low cytotoxic capacity, produces higher amounts of interferon (IFN)-γ, and expresses functional homologs of human NK cell immunoglobulin-like receptors. We further show that bone marrow-derived CD27(low) mNK cells are selectively recruited to the uterus and ameliorate the rate of fetal loss when adoptively transferred into alymphoid RAG2(-/-)/γc(-/-) mice. Additionally, expression of CD27 is down-modulated on mNK cells upon migration to the uterus. Hence, we propose the existence of a regulatory mNK cell subset, which is licensed toward successful pregnancy maintenance at the fetomaternal interface in mice. As CD27(low) NK cells are also present in human decidua, the CD27(low) NK subset may provide a tool to foster translational research in reproduction, aiming to improve pregnancy outcome in humans.

Coronary CT angiography (CCTA) has the advantage over invasive coronary angiography in that its non-invasive nature and minimal risk on patients. CCTA enables accurate assessment of the entire heart, coronary artery system and thorax, displaying three-dimensional information about the spatial relations of the anomalous vessels and surrounding intraluminal and extraluminal anatomy, and thereby contributing clinically important prognostic information. Dual left anterior descending (LAD) coronary artery consists of of two LAD arteries within the anterior interventricular sulcus (AIVS). Type 4 is infrequently reported subtype and differs from the others, with a long LAD originating from the right coronary artery (Mercado, A., Johnson Jr, G., Calver, D., & Sokol, R. J. (1989). Cocaine, pregnancy, and postpartum intracerebral hemorrhage. Obstetrics & Gynecology, 73(3, Part 2), 467-468. and the short LAD originating from the left main coronary artery. However, the radiological features between the short LAD and septal coronary arteries remain a controversy, with the latter being determined by CCTA. We present a case report based on short LAD terminating proximally in the AIVS and the long LAD originating from the RCA and terminating into the distal AIVS with the later having a long septal travelling parallel to the long LAD.

The crosstalk between NK cells and M1 macrophages has a vital role in the protection against infections and tumor development. However, macrophages in the tumor resemble an M2 phenotype, and, at present, their effect on NK cells is less clear. This study investigated whether tumor-associated macrophages (TAMs) have a role in altering NK cell function and phenotype using in vitro cocultures of murine NK cells with peritoneal or bone marrow-derived, M2-polarized macrophages or TAMs isolated from spontaneous mouse breast tumors. We report here that both peritoneal and bone marrow-derived M2 macrophages, as well as TAMs, substantially inhibit NK cell activation and concordant cytotoxicity against tumor cells. The mechanism for this inhibition was found to require contact between the respective cell types. Both M2 macrophages and TAMs are producers of the immunosuppressive cytokine TGF-β. The inhibition of TGF-β restored the cytotoxicity of NK cells in contact with M2 macrophages, implicating TGF-β in the mechanism for NK cell inhibition. In addition to affecting NK cell function, TAMs also induced a CD27lowCD11bhigh-exhausted NK cell phenotype, which corresponds with the reduced activation and cytotoxicity observed. This study reveals a novel implication of TAMs in the tumor-associated inhibition of NK cell function by demonstrating their capacity to directly alter NK cell cytotoxicity and phenotype in a contact-dependent mechanism involving TGF-β. These findings identify the interaction between NK cells and TAMs as a prospective therapeutic target to enhance NK cell effector function for effective NK cell cancer therapies.

Objective:To investigate the short term effect of hubble-bubble smoking on voice.Study design:Prospective study.Material:Eighteen non-dysphonic subjects (seven men and 11 women) with a history of hubble-bubble smoking and no history of cigarette smoking underwent acoustic analysis and laryngeal video-stroboscopic examination before and 30 minutes after hubble-bubble smoking.Results:On laryngeal video-stroboscopy, none of the subjects had vocal fold erythema either before or after smoking. Five patients had mild vocal fold oedema both before and after smoking. After smoking, there was a slight increase in the number of subjects with thick mucus between the vocal folds (six, vs four before smoking) and with vocal fold vessel dilation (two, vs one before smoking). Acoustic analysis indicated a drop in habitual pitch, fundamental frequency and voice turbulence index after smoking, and an increase in noise-to-harmonics ratio.Conclusion:Even 30 minutes of hubble-bubble smoking can cause a drop in vocal pitch and an increase in laryngeal secretions and vocal fold vasodilation.

Natural Killer cells can distinguish between healthy and malignant cells and have the unique ability to lyse tumour cells without prior sensitization. Differences between murine and human NK cells complicate the translation of this knowledge into useful therapeutics. Humanized mouse models that support the development of human leukocytes are a promising avenue of research that aims to address this problem. Here we provide an in-depth phenotypic analysis of human NK cells in Balb/c Rag2(-/-)γ(c)(-/-) mice reconstituted with human hematopoietic stem cells. We have examined the development of NK cells in bone marrow, thymous, spleen, lymph node (LN) and liver. Interestingly, in naive reconstituted mice, NK cells were found in thymus and LN, but not in bone marrow. These NK cells expressed several inhibitory and activating receptors needed for malignant cell detection. Furthermore, we confirm that administration of recombinant human interleukin-15 (rhIL-15) or Ad-vector expressing hIL-15 is able to significantly enhance NK cell development and maturation, particularly in bone marrow and liver, in this model. Our results suggest that human NK cells developed in mice may have phenotypes and tissue distributions similar to those seen in human.