A polymorphism in IRF4 affects human pigmentation through a tyrosinase-dependent MITF/TFAP2A pathway.

Sequence polymorphisms linked to human diseases and phenotypes in genome-wide association studies often affect noncoding regions. A SNP within an intron of the gene encoding Interferon Regulatory Factor 4 (IRF4), a transcription factor with no known role in melanocyte biology, is strongly associated with sensitivity of skin to sun exposure, freckles, blue eyes, and brown hair color. Here, we demonstrate that this SNP lies within an enhancer of IRF4 transcription in melanocytes. The allele associated with this pigmentation phenotype impairs binding of the TFAP2A transcription factor that, together with the melanocyte master regulator MITF, regulates activity of the enhancer. Assays in zebrafish and mice reveal that IRF4 cooperates with MITF to activate expression of Tyrosinase (TYR), an essential enzyme in melanin synthesis. Our findings provide a clear example of a noncoding polymorphism that affects a phenotype by modulating a developmental gene regulatory network.

Suppression of Ca2+ signaling enhances melanoma progression.

The role of store-operated Ca2+ entry (SOCE) in melanoma metastasis is highly controversial. To address this, we here examined UV-dependent metastasis, revealing a critical role for SOCE suppression in melanoma progression. UV-induced cholesterol biosynthesis was critical for UV-induced SOCE suppression and subsequent metastasis, although SOCE suppression alone was both necessary and sufficient for metastasis to occur. Further, SOCE suppression was responsible for UV-dependent differences in gene expression associated with both increased invasion and reduced glucose metabolism. Functional analyses further established that increased glucose uptake leads to a metabolic shift towards biosynthetic pathways critical for melanoma metastasis. Finally, examination of fresh surgically isolated human melanoma explants revealed cholesterol biosynthesis-dependent reduced SOCE. Invasiveness could be reversed with either cholesterol biosynthesis inhibitors or pharmacological SOCE potentiation. Collectively, we provide evidence that, contrary to current thinking, Ca2+ signals can block invasive behavior, and suppression of these signals promotes invasion and metastasis.

Suppression of Ca2+ signals by EGR4 controls Th1 differentiation and anti-cancer immunity in vivo.

While the zinc finger transcription factors EGR1, EGR2, and EGR3 are recognized as critical for T-cell function, the role of EGR4 remains unstudied. Here, we show that EGR4 is rapidly upregulated upon TCR engagement, serving as a critical "brake" on T-cell activation. Hence, TCR engagement of EGR4-/- T cells leads to enhanced Ca2+ responses, driving sustained NFAT activation and hyperproliferation. This causes profound increases in IFNγ production under resting and diverse polarizing conditions that could be reversed by pharmacological attenuation of Ca2+ entry. Finally, an in vivo melanoma lung colonization assay reveals enhanced anti-tumor immunity in EGR4-/- mice, attributable to Th1 bias, Treg loss, and increased CTL generation in the tumor microenvironment. Overall, these observations reveal for the first time that EGR4 is a key regulator of T-cell differentiation and function.

Gadd45 in Senescence.

Gadd45a, Gadd45b, and Gadd45g have been implicated in cell cycle arrest, DNA repair, apoptosis, innate immunity, genomic stability, and more recently in senescence. Evidence has accumulated that Gadd45a deficiency results in escape of mouse embryo fibroblasts from senescence, whereas Gadd45b deficiency promotes premature senescence and skin aging. Moreover, recently Gadd45b deficiency was found to promote senescence and attenuate liver fibrosis, whereas Gadd45a was observed to exert a protective effect against hepatic fibrosis. These findings indicate that the Gadd45 stress response proteins play important roles in modulating cellular responses to senescence. Thus, exploring how Gadd45 proteins modulate cellular senescence has the potential to provide new and innovative tools to treat cancer as well as liver disease.

Lead in the marine environment: concentrations and effects on invertebrates.

Lead (Pb) is a non-essential metal naturally present in the environment and often complexed with other elements (e.g., copper, selenium, zinc). This metal has been used since ancient Egypt and its extraction has grown in the last centuries. It has been used until recently as a fuel additive and is currently used in the production of vehicle batteries, paint, and plumbing. Marine ecosystems are sinks of terrestrial contaminations; consequently, lead is detected in oceans and seas. Furthermore, lead is not biodegradable. It remains in soil, atmosphere, and water inducing multiple negative impacts on marine invertebrates (key species in trophic chain) disturbing ecological ecosystems. This review established our knowledge on lead accumulation and its effects on marine invertebrates (Annelida, Cnidaria, Crustacea, Echinodermata, and Mollusca). Lead may affect different stages of development from fertilization to larval development and can also lead to disturbance in reproduction and mortality. Furthermore, we discussed changes in the seawater chemistry due to Ocean Acidification, which can affect the solubility, speciation, and distribution of the lead, increasing potentially its toxicity to marine invertebrates.

Programming of donor T cells using allogeneic δ-like ligand 4-positive dendritic cells to reduce GVHD in mice.

Alloreactive T cells play a critical role in eliminating hematopoietic malignant cells but are also the mediators of graft-versus-host disease (GVHD), a major complication that subverts the success of allogeneic hematopoietic stem cell transplantation (HSCT). However, induction of alloreactive T cells does not necessarily lead to GVHD. Here we report the development of a cellular programming approach to render alloreactive T cells incapable of causing severe GVHD in both major histocompatibility complex (MHC)-mismatched and MHC-identical but minor histocompatibility antigen-mismatched mouse models. We established a novel platform that produced δ-like ligand 4-positive dendritic cells (Dll4(hi)DCs) from murine bone marrow using Flt3 ligand and Toll-like receptor agonists. Upon allogeneic Dll4(hi)DC stimulation, CD4(+) naïve T cells underwent effector differentiation and produced high levels of interferon γ (IFN-γ) and interleukin-17 in vitro, depending on Dll4 activation of Notch signaling. Following transfer, allogeneic Dll4(hi)DC-induced T cells were unable to mediate severe GVHD but preserved antileukemic activity, significantly improving the survival of leukemic mice undergoing allogeneic HSCT. This effect of Dll4(hi)DC-induced T cells was associated with their impaired expansion in GVHD target tissues. IFN-γ was important for Dll4(hi)DC programming to reduce GVHD toxicities of alloreactive T cells. Absence of T-cell IFN-γ led to improved survival and expansion of Dll4(hi)DC-induced CD4(+) T cells in transplant recipients and caused lethal GVHD. Our findings demonstrate that Dll4(hi)DC programming can overcome GVHD toxicity of donor T cells and produce leukemia-reactive T cells for effective immunotherapy.

Spatiotemporal Labeling of Melanocytes in Mice.

Melanocytes are pigment producing cells in the skin that give rise to cutaneous malignant melanoma, which is a highly aggressive and the deadliest form of skin cancer. Studying melanocytes in vivo is often difficult due to their small proportion in the skin and the lack of specific cell surface markers. Several genetically-engineered mouse models (GEMMs) have been created to specifically label the melanocyte compartment. These models give both spatial and temporal control over the expression of a cellular 'beacon' that has an added benefit of inducible expression that can be activated on demand. Two powerful models that are discussed in this review include the melanocyte-specific, tetracycline-inducible green fluorescent protein expression system (iDct-GFP), and the fluorescent ubiquitination-based cell cycle indicator (FUCCI) model that allows for the monitoring of the cell-cycle. These two systems are powerful tools in studying melanocyte and melanoma biology. We discuss their current uses and how they could be employed to help answer unresolved questions in the fields of melanocyte and melanoma biology.

Genetically engineered mouse models of melanoma.

Melanoma is a complex disease that exhibits highly heterogeneous etiological, histopathological, and genetic features, as well as therapeutic responses. Genetically engineered mouse (GEM) models provide powerful tools to unravel the molecular mechanisms critical for melanoma development and drug resistance. Here, we expound briefly the basis of the mouse modeling design, the available technology for genetic engineering, and the aspects influencing the use of GEMs to model melanoma. Furthermore, we describe in detail the currently available GEM models of melanoma. Cancer 2017;123:2089-103. © 2017 American Cancer Society.

TAMeless traitors: macrophages in cancer progression and metastasis.

Macrophages are conventionally classified into M1 and M2 subtypes according to their differentiation status and functional role in the immune system. However, accumulating evidence suggests that this binary classification system is insufficient to account for the remarkable plasticity of macrophages that gives rise to an immense diversity of subtypes. This diverse spectrum of macrophage subtypes play critical roles in various homeostatic and immune functions, but remain far from being fully characterised. In addition to their roles in normal physiological conditions, macrophages also play crucial roles in disease conditions such as cancer. In this review, we discuss the roles tumour-associated macrophages (TAMs) play in regulating different steps of tumour progression and metastasis, and the opportunities to target them in the quest for cancer prevention and treatment.

Interferon-γ links ultraviolet radiation to melanomagenesis in mice.

Cutaneous malignant melanoma is a highly aggressive and frequently chemoresistant cancer, the incidence of which continues to rise. Epidemiological studies show that the major aetiological melanoma risk factor is ultraviolet (UV) solar radiation, with the highest risk associated with intermittent burning doses, especially during childhood. We have experimentally validated these epidemiological findings using the hepatocyte growth factor/scatter factor transgenic mouse model, which develops lesions in stages highly reminiscent of human melanoma with respect to biological, genetic and aetiological criteria, but only when irradiated as neonatal pups with UVB, not UVA. However, the mechanisms underlying UVB-initiated, neonatal-specific melanomagenesis remain largely unknown. Here we introduce a mouse model permitting fluorescence-aided melanocyte imaging and isolation following in vivo UV irradiation. We use expression profiling to show that activated neonatal skin melanocytes isolated following a melanomagenic UVB dose bear a distinct, persistent interferon response signature, including genes associated with immunoevasion. UVB-induced melanocyte activation, characterized by aberrant growth and migration, was abolished by antibody-mediated systemic blockade of interferon-γ (IFN-γ), but not type-I interferons. IFN-γ was produced by macrophages recruited to neonatal skin by UVB-induced ligands to the chemokine receptor Ccr2. Admixed recruited skin macrophages enhanced transplanted melanoma growth by inhibiting apoptosis; notably, IFN-γ blockade abolished macrophage-enhanced melanoma growth and survival. IFN-γ-producing macrophages were also identified in 70% of human melanomas examined. Our data reveal an unanticipated role for IFN-γ in promoting melanocytic cell survival/immunoevasion, identifying a novel candidate therapeutic target for a subset of melanoma patients.

A novel interaction between the FLJ33534 locus and smoking in obesity: a genome-wide study of 14 131 Pakistani adults.

BACKGROUND: Obesity is a complex disease caused by the interplay of genetic and lifestyle factors, but identification of gene-lifestyle interactions in obesity has remained challenging. Few large-scale studies have reported use of genome-wide approaches to investigate gene-lifestyle interactions in obesity. METHODS: In the Pakistan Risk of Myocardial Infraction Study, a cross-sectional study based in Pakistan, we calculated body mass index (BMI) variance estimates (square of the residual of inverse-normal transformed BMI z-score) in 14 131 participants and conducted genome-wide heterogeneity of variance analyses (GWHVA) for this outcome. All analyses were adjusted for age, age(2), sex and genetic ancestry. RESULTS: The GWHVA analyses identified an intronic variant, rs140133294, in the FLJ33544 gene in association with BMI variance (P-value=3.1 × 10(-8)). In explicit tests of gene × lifestyle interaction, smoking was found to significantly modify the effect of rs140133294 on BMI (Pinteraction=0.0005), whereby the minor allele (T) was associated with lower BMI in current smokers, while positively associated with BMI in never smokers. Analyses of ENCODE data at the FLJ33534 locus revealed features indicative of open chromatin and high confidence DNA-binding motifs for several transcription factors, providing suggestive biological support for a mechanism of interaction. CONCLUSIONS: In summary, we have identified a novel interaction between smoking and variation at the FLJ33534 locus in relation to BMI in people from Pakistan.

Comparative e-waste plastics biodegradation efficacy of monoculture Pseudomonas aeruginosa strain PE10 and bacterial consortium under in situ condition.

A significant amount of electronic obsoletes or electronic waste (e-waste) is being generated globally each year; of these, ~20% of obsolete electronic items have plastic components. Current remediation practices for e-waste have several setbacks due to its negative impact on the environment, agro-ecosystem, and human health. Therefore, comparative biodegradation studies of e-waste plastics by monoculture Pseudomonas aeruginosa strain PE10 and bacterial consortium consisting of Achromobacter insolitus strain PE2 (MF943156), Acinetobacter nosocomialis strain PE5 (MF943157), Pseudomonas lalkuanensis PE8 (CP043311), and Stenotrophomonas pavanii strain PE15 (MF943160) were carried out in situ. Biological treatment of e-waste with these candidates in soil ecosystems has been analyzed through diversified analytical techniques such as Fourier transform infrared spectroscopy (FTIR), thermogravimetric-derivative thermogravimetry-differential thermal analysis (TG-DTG-DTA), and scanning electron microscopy (SEM). Both P. aeruginosa strain PE10 and the bacterial consortium have a tremendous ability to accelerate the biodegradation process in the natural environment. However, FTIR analysis implied that the monoculture had better efficacy than the consortium, and it was consistent until the incubation period used for the study. Some polymeric bonds such as ν C=C and δ C-H were completely removed, and ν C=C ring stretching, νasym C-O-C, νsym C-H, etc. were introduced by strain PE10. Furthermore, thermal analysis results validated the structural deterioration of e-waste as the treated samples showed nearly two-fold weight loss (WL; 6.8%) than the untreated control (3.1%) at comparatively lower temperatures. SEM images provided the details of surface disintegrations. Conclusively, individual monoculture P. aeruginosa strain PE10 could be explored for e-waste bio-recycling in agricultural soil ecosystems thereby reducing the cost, time, and management of bioformulation in addition to hazardous pollutant reduction.

Interferon-gamma signaling promotes melanoma progression and metastasis.

Interferon-gamma (IFNG) has long been regarded as the flag-bearer for the anti-cancer immunosurveillance mechanisms. However, relatively recent studies have suggested a dual role of IFNG, albeit there is no direct experimental evidence for its potential pro-tumor functions. Here we provide in vivo evidence that treatment of mouse melanoma cell lines with Ifng enhances their tumorigenicity and metastasis in lung colonization allograft assays performed in immunocompetent syngeneic host mice, but not in immunocompromised host mice. We also show that this enhancement is dependent on downstream signaling via Stat1 but not Stat3, suggesting an oncogenic function of Stat1 in melanoma. The experimental results suggest that melanoma cell-specific Ifng signaling modulates the tumor microenvironment and its pro-tumorigenic effects are partially dependent on the γδ T cells, as Ifng-enhanced tumorigenesis was inhibited in the TCR-δ knockout mice. Overall, these results show that Ifng signaling may have tumor-promoting effects in melanoma by modulating the immune cell composition of the tumor microenvironment.

Electrochemical monitoring of congo red degradation using strontium titanate-doped biochar nanohybrids derived photocatalytic plates.

Present investigation demonstrates the development and characterization of strontium titanate (SrTiO3) doped biochar nanohybrid photocatalysts. Biochar nanohybrid was synthesized using an ultrasonic-assisted dispersion technique, which involved dispersing SrTiO3 nanoparticles into activated biochar at a weight ratio of 1:2 (w/w) under ambient conditions. The development of the biochar nanohybrid was verified through a comprehensive analysis of their spectral, microstructural, thermal, electrical, and electrochemical properties. The scanning electron microscopy analysis reveals a surface-associated multiphase morphology of the biochar nanohybrid, attributed to the uniform distribution of SrTiO3 within the activated biochar matrix. Biochar nanohybrid exhibited a reduced optical band gap of 2.77 eV, accompanied by a crystallite size of 32.45. Thermogravimetric analysis revealed the thermal stability of the biochar nanohybrid, as evidenced by a char residue of 70.83% at 1000 °C. The working electrodes derived from biochar nanohybrid have exhibited ohmic behavior and displayed a significantly enhanced DC conductivity (mS/cm) of 1.13, surpassing that of activated biochar (0.53) and SrTiO3 (0.62) at 100 V. The developed biochar nanohybrid were employed for the degradation of congo red dye by exposing the dye solution to photocatalytic plates. These photocatalytic plates were prepared by coating biochar nanohybrid onto glass plates using epoxy-based reactive binders for secure binding. The photodegradation of congo red was evaluated through cyclic voltammetric analysis in a 0.1 M KCl solution at pH 8.0, resulting in an impressive 99.95% photocatalytic efficiency in degrading a congo red solution (50 mg/L). This study presents a novel approach for the fabrication of biochar nanohybrid-derived photocatalytic plates, offering high photocatalytic efficiency for the degradation of congo red dye.

Advanced breast conservation and partial breast reconstruction - a review of current available options for oncoplastic breast surgery.

Breast cancer is the most common cancer affecting one in three women with new cancer diagnosis in England. Breast-conserving surgery is the primary surgical option in a vast majority of these patients. Use of oncoplastic techniques in breast conservation surgery has significantly improved the aesthetic outcomes without compromising the oncological safety of cancer resections. Oncoplastic breast-conserving surgery (OPBCS) has transformed the specialty with a paradigm shift in ideology and the recognition that aesthetic and oncological resections are synonymous when planning surgical intervention for patients with breast cancer. The two main options for OPBCS are therapeutic mammoplasty and partial beast reconstruction using pedicle-based flaps. This review aims to highlight key concepts in OPBCS demonstrating an overview of these surgical techniques, their safety, outcomes and the emergence of extreme oncoplastic breast surgery.

Ultrasound-positive axillary lymph nodes in early breast cancer: can a patient subgroup avoid axillary lymph node dissection? A simple algorithm is suggested.

INTRODUCTION: National UK guidelines suggest that axillary lymph node dissection (ALND) is no longer mandatory for selected early node-positive breast cancer patients. Our study aimed to identify patients with early breast cancer and ultrasound (USS)-positive axillary metastasis who possess low burden of axillary disease and can avoid ALND. METHODS: We conducted a 5-year study of prospectively collected data of patients with clinically T1-2, N0 breast cancer and a positive USS-guided axillary biopsy. Primary outcome was involvement of 1-2 lymph nodes (low disease burden) or ≥3 lymph nodes (higher axillary disease) on final ALND histology. Tumour type, size, grade, multifocality, receptor status, number of abnormal imaged nodes and presence of lympho-vascular invasion (LVI) were recorded. Data were analysed using chi-squared and Student's t-test. RESULTS: One hundred and sixty-six patients underwent ALND for pT1-2 breast cancer. Seventy patients had no clinically palpable lymphadenopathy but a positive USS-guided biopsy. Of 70 patients, 32 women (46%) had low disease burden, whereas 38 women (54%) had higher axillary disease in final histology. LVI and >1 abnormal lymph node on USS were both significantly associated with higher disease burden (p = 0.050 and 0.009, respectively). CONCLUSION: Our study confirms the presence of an important patient cohort, who are clinically node-negative with a positive USS-guided biopsy and a low volume of axillary disease. No imaging modality currently has the accuracy required to identify patients with this low disease burden preoperatively but we propose a simple algorithm for axillary management in this subgroup.

Interferon-gamma induces melanogenesis via post-translational regulation of tyrosinase.

Melanogenesis (melanin pigment production) in melanocytes is canonically stimulated by the alpha melanocyte stimulating hormone (αMSH), which activates the cyclic-AMP-mediated expression of the melanocyte inducing transcription factor (MITF) and its downstream melanogenic genes, including the principal rate-limiting melanogenic enzyme tyrosinase (TYR). Here, we report that interferon-gamma (IFNG; type II interferon), but not interferon-alpha (a type I interferon), induces a noncanonical melanogenic pathway in mouse and human melanocytic cells. Inhibition of IFNG pathway by the JAK1/2 inhibitor ruxolitinib or knocking out Stat1 gene abrogated the IFNG-induced melanogenesis. Interestingly, IFNG-induced melanogenesis was independent of MITF. IFNG markedly increased the TYR protein expression but did not affect the mRNA expression, suggesting a post-translational regulatory mechanism. In contrast, IFNG had no effect on the expression of other melanogenesis-related proteins, for example, tyrosinase-related protein 1 (TYRP1) and dopachrome tautomerase (DCT). Glycosidase digestion assays revealed that IFNG treatment increased the mature glycosylated form of TYR, but not its de novo synthesis. Moreover, cycloheximide chase assay showed that degradation of TYR was decreased in IFNG-treated cells. These results suggest that the IFNG-STAT1 pathway regulates melanogenesis via regulation of the post-translational processing and protein stability of TYR.

A new function for CD38/ADP-ribosyl cyclase in nuclear Ca2+ homeostasis.

Nucleoplasmic calcium ions (Ca2+) influence nuclear functions as critical as gene transcription, apoptosis, DNA repair, topoisomerase activation and polymerase unfolding. Although both inositol trisphosphate receptors and ryanodine receptors, types of Ca2+ channel, are present in the nuclear membrane, their role in the homeostasis of nuclear Ca2+ remains unclear. Here we report the existence in the inner nuclear membrane of a functionally active CD38/ADP-ribosyl cyclase that has its catalytic site within the nucleoplasm. We propose that the enzyme catalyses the intranuclear cyclization of nicotinamide adenine dinucleotide to cyclic adenosine diphosphate ribose. The latter activates ryanodine receptors of the inner nuclear membrane to trigger nucleoplasmic Ca2+ release.

Implications of SPION and NBT nanoparticles upon in-vitro and in-situ biodegradation of LDPE film.

Comparative influence of two nanoparticles viz. superparamagnetic iron oxide nanoparticles (SPION) and nanobarium titanate (NBT) was studied upon the in-vitro and in-situ low-density polyethylene (LDPE) biodegradation efficiency of a potential polymer-degrading microbial consortium. Supplementation of 0.01% concentration (w/v) of the nanoparticles in minimal broth significantly increased the bacterial growth, along with early onset of the exponential phase. Under in-vitro conditions, lambda-max shifts were quicker with nanoparticles and Fourier transform infrared spectroscopy (FTIR) illustrated significant changes in CH/CH2 vibrations, along with introduction of hydroxyl residues in the polymer backbone. Further, simultaneous thermogravimetric-differential thermogravimetry-differential thermal analysis (TG-DTG-DTA) reported multiple-step decomposition of LDPE degraded in the presence of nanoparticles. These findings were supported by scanning electron micrographs (SEM) which revealed greater dissolution of film surface in the presence of nanoparticles. Furthermore, progressive degradation of the film was greatly enhanced when it was incubated under soil conditions for 3 months with the nanoparticles. The study highlights the significance of bacteria-nanoparticle interactions which can dramatically influence key metabolic processes like biodegradation. The authors also propose the exploration of nanoparticles to influence various other microbial processes for commercial viabilities.