Targeted autophagy disruption reveals the central role of macrophage iron metabolism in systemic iron homeostasis.

Iron homeostasis depends on both intracellular control through iron-responsive proteins and the systemic level of iron through hepcidin-ferroportin axis. Indeed, the hormone hepcidin downregulates the ferroportin iron exporter to control iron recycling from macrophages and iron uptake from enterocytes. Here, we focused on the role of autophagy in macrophage iron metabolism and systemic iron homeostasis. Mice deficient for autophagy in macrophages (LysM-Atg5-/-) mimicked a primary iron overload phenotype, resulting in high ferroportin expression in both macrophages and enterocytes that correlated with marked parenchymal iron overload. Furthermore, LysM-Atg5-/- mice exhibited increased hematopoietic activity with no sign of anemia but correlating with rather high plasma iron level. Compared with wild-type cells, bone marrow-derived macrophages from LysM-Atg5-/- mice had significantly increased ferroportin expression and decreased iron content, confirming high iron export. In erythrophagocytic macrophages, autophagy regulates hemosiderin storage mechanisms as well as degradation of ferroportin and subsequently its plasma membrane localization and iron export; furthermore, ferroportin colocalization with hepcidin indicates hepcidin autocrine activity. Relatively high hepatic hepcidin expression and decreased hepcidin level in the spleen of LysM-Atg5-/- mice, correlating with low hemosiderin iron storage, as well as in erythrophagocytic Atg5-/- macrophages were evidenced. Therefore, our results highlight the critical role of autophagy in macrophages for iron trafficking and systemic iron homeostasis. We propose that in macrophages, autophagy restricts ferroportin level and iron export, resulting in hepcidin expression with an autocrine-paracrine effect that plays a role in the regulation of ferroportin expression in duodenal enterocytes.

Effectiveness of broad-spectrum antiseptics in production of disinfected maggots of Lucilia sericata for use in wound debridement therapy.

The establishment of low-cost, effective, safe and practical methods is necessary to increase the use of larval therapy in wound care. Although studies on external disinfection of calliphorid eggs have been reported, many studies lack data on the effect of disinfection on egg viability and the microorganisms found before disinfection. Therefore, the main objective of this study was to compare three antiseptic solutions, that is, chlorhexidine (5%), Dakin's solution (0.5% NaOCl) and povidone-iodine (10%), in terms of their ability to disinfect Lucilia sericata eggs. Egg viability after disinfection and microorganisms present on the eggs and larvae before and after treatment were also examined. None of the antiseptics had a significant effect on egg viability. Disinfection of L. sericata eggs with 0.5% NaOCl was the best method, as sterility tests showed no contamination. Escherichia coli, Bacillus subtilis and Proteus mirabilis were present in all cultures isolated from the non-disinfected eggs and larvae, while Enterococcus faecium, Enterococcus faecalis, Morganella morganii, Corynebacterium spp. and Providencia stuartii were isolated from more than half of the same cultures. Sterility testing of medicinal maggots after disinfection is crucial to prevent secondary infections and achieve a positive therapeutic outcome.

DNA barcoding of Stearibia nigriceps (Meigen) and Piophila casei (Linnaeus) (Diptera: Piophilidae) from Algeria and the first African report of Stearibia nigriceps.

Stearibia (=Piophila) nigriceps (Meigen) occurs in the Palearctic Region from Europe and Far East Russia, and in the Nearctic, the Neotropical, and the Oriental regions. In this study, we report the first record of Stearibia nigriceps from Africa collected in Algeria. In addition, our results provide DNA sequences of Piophila casei (Linnaeus) and S. nigriceps that may serve as reference data for future identification. A region of the cytochrome c oxidase I (COI) gene marker of 473 bp was amplified and DNA was sequenced. Intra- and interspecific genetic distances were calculated. A phylogenetic tree was generated by the maximum likelihood (ML) method using 1000 bootstrap replicates based on the Tamura-Nei model. A total number of 9 adults of S. nigriceps were collected together with 72 adults of P. casei. The intraspecific variation of the species barcodes analysed in our study was < 3% and the interspecific nucleotide divergence of the same species was > 3%. The 473 bp COI region was sufficient to identify and distinguish unambiguously between the two species. P. casei and P. nigriceps individuals from Algeria clustered separately from each other. They were also separated from the individuals of the same species originating from other countries with a 100% high bootstrap value. There was no correlation between the clades and the geographic origins of the haplotypes. Our results contribute to update the knowledge of the distribution of Piophilidae family. They also contribute to the build-up of online DNA sequence databases.

c-Kit signaling potentiates CAR T cell efficacy in solid tumors by CD28- and IL-2-independent co-stimulation.

The limited efficacy of chimeric antigen receptor (CAR) T cell therapy for solid tumors necessitates engineering strategies that promote functional persistence in an immunosuppressive environment. Herein, we use c-Kit signaling, a physiological pathway associated with stemness in hematopoietic progenitor cells (T cells lose expression of c-Kit during differentiation). CAR T cells with intracellular expression, but no cell-surface receptor expression, of the c-Kit D816V mutation (KITv) have upregulated STAT phosphorylation, antigen activation-dependent proliferation and CD28- and interleukin-2-independent and interferon-γ-mediated co-stimulation, augmenting the cytotoxicity of first-generation CAR T cells. This translates to enhanced survival, including in transforming growth factor-ß-rich and low-antigen-expressing solid tumor models. KITv CAR T cells have equivalent or better in vivo efficacy than second-generation CAR T cells and are susceptible to tyrosine kinase inhibitors (safety switch). When combined with CD28 co-stimulation, KITv co-stimulation functions as a third signal, enhancing efficacy and providing a potent approach to treat solid tumors.

Impact of plastic wrapping on carcass decomposition and arthropod colonisation in northern Africa during spring.

The effect of plastic wrapping on decomposition rate and carrion fauna of rabbits (Oryctolagus cuniculus L.) was examined in spring in a semi-urban area in North Algeria. All decomposition stages were observed in all carcasses, with the same durations in the control but different durations in the wrapped carcasses. Decomposition of the carcasses in the plastic wrapping was significantly slower than that of the exposed ones. A total of 12,516 specimens, belonging to 36 families and 69 species, were morphologically identified. Thirteen species of forensic relevance were also identified at the molecular level using the cytochrome c oxidase I (COI) barcode region, and the sequences were submitted to online databases. Wrapping had a significant effect on species composition (χ2 = 569.269, df = 55, p < 0.001). Higher species richness, abundance, and diversity were found in the control group. No significant difference in species abundance was observed between the treatments. The plastic wrap did not influence the accessibility of carcasses to insects, nor did it delay the arrival of necrophagous flies. This study provides basic information on the decomposition and arthropod colonisation of wrapped remains and contributes to the literature on North African carrion fauna.

Molecular identification of the potentially forensically relevant cluster flies Pollenia rudis (Fabricius) and Pollenia vagabunda (Meigen) (Diptera: Polleniidae) - non-recorded species in Algeria.

Cluster flies are represented by the genus Pollenia Robineau-Desvoidy, 1830 of the family Polleniidae Brauer and Bergenstamm, 1889. Their larvae are known to be internal parasites or predators of earthworms. Herein, we report for the first time the occurrence of the cluster flies Pollenia rudis Fabricius, 1794 and Pollenia vagabunda (Meigen, 1826) (Diptera: Polleniidae) on carcasses in Algeria and identify them through DNA barcoding. A region of the mitochondrial cytochrome c oxidase I gene (COI) was amplified and sequenced. Genetic distances were determined. A phylogenetic tree was constructed with the maximum parsimony method using 10 000 bootstrap replicates. A total number of 157 adults of P. rudis were collected together with 325 adults of Pollenia vagabunda. The occurrence of Pollenia on animal carcasses does not seem to be correlated with a particular stage of decomposition. All the sequences were correctly identified using the BLASTn tool from the GenBank database and the BOLD identification engine. Intra- and interspecific sequence divergence values were less than 1% and greater than 3%, respectively. COI barcodes obtained from this study were robust enough to identify and distinguish unambiguously between P. rudis and P. vagabunda. In the tree-based analysis, the cluster flies were all assigned to their respective species separately from each other confirming the morphological identification. These results provide DNA barcodes that contribute to the growth of reference databases and allow fast and accurate identification.

Assessment of Entomological Remains from Soil Samples Collected from a Pig (Sus scrofa domestica) Carcass Decomposition Site after 13 Years.

OBJECTIVE: Carrion insects inhabiting the soil play an important role in forensic investigations because they may help to solve both active and cold cases. The aim of this study was to examine the entomofauna of forensic importance in soil samples removed after 13 years from a pig carrion decomposition site. METHODS: Soil samples were collected from an old carrion decomposition study site in Bâla, the Ankara Province. Four holes, approximately 40 cm deep and 35 cm width were excavated at the study site. The samples were collected and placed in ventilated cups. Each cup was labeled mentioning the excavation location, time, date, and name of the collector. Insects and their remains found in the soil were collected by sweeping the soil from the specimens using a brush. The insects were morphologically identified. RESULTS: A total of 635 specimens of Calliphoridae, Dermestidae, Cleridae, Staphylinidae, Histeridae, and Formicidae were identified. Flies such as Chrysomya albiceps (Wiedmann, 1819), and beetles such as Dermestes frischii (Kugelan, 1792), Necrobia rufipes (De Geer, 1775), and Creophilus maxillosus (Linnaeus, 1758), were identified as the species. CONCLUSION: Our results show that soil samples still harbor entomological specimens after 13 years. This study, to the best of our knowledge, was the first of its kind in Turkey. Forensically, important insects and their remains may be identified in the soil long time after the corpse is buried. Consequently, cold cases may be solved using insects.

Cell iron status influences macrophage polarization.

Macrophages play crucial roles in innate immune response and in the priming of adaptive immunity, and are characterized by their phenotypic heterogeneity and plasticity. Reprogramming intracellular metabolism in response to microenvironmental signals is required for M1/M2 macrophage polarization and function. Here we assessed the influence of iron on the polarization of the immune response in vivo and in vitro. Iron-enriched diet increased M2 marker Arg1 and Ym1 expression in liver and peritoneal macrophages, while iron deficiency decreased Arg1 expression. Under LPS-induced inflammatory conditions, low iron diet exacerbated the proinflammatory response, while the IL-12/IL-10 balance decreased with iron-rich diet, thus polarizing toward type 2 response. Indeed, in vitro macrophage iron loading reduced the basal percentage of cells expressing M1 co-stimulatory CD86 and MHC-II molecules. Further, iron loading of macrophages prevented the pro-inflammatory response induced by LPS through reduction of NF-κB p65 nuclear translocation with decreased iNOS, IL-1ß, IL-6, IL-12 and TNFα expression. The increase of intracellular iron also reduced LPS-induced hepcidin gene expression and abolished ferroportin down-regulation in macrophages, in line with macrophage polarization. Thus, iron modulates the inflammatory response outcome, as elevated iron levels increased M2 phenotype and negatively regulated M1 proinflammatory LPS-induced response.

Small cell carcinoma of the urinary bladder: a case report and review of the literature.

Small cell carcinoma of the bladder (SCCB) is extremely rare. In this paper, we present a case of metastatic SCCB managed by chemotherapy and we would provide a brief review of the epidemiology, clinical features, diagnosis, pathologic features, staging, treatment, and prognosis of SCCB. A 52-year-old man was admitted with signs and symptoms suggestive of a bladder cancer. Computed tomography of the pelvis and abdomen showed a large tumor at the right bladder wall, measuring 10 cm in diameter, and a multinodular liver disease. Diagnosis of small cell carcinoma was established from the histological study of the transurethral resection of the bladder tumor. The patient received 12 cycles of platinum-based chemotherapy with a good partial response of bladder tumor and liver metastasis. The patient is still alive, 18 months after diagnosis.