Long Glucocorticoid-induced Leucine Zipper (L-GILZ) è essenziale per il controllo della spermatogenesi

A correct balance between spermatogonial stem cells (SSCs) self-renewal and differentiation is essential for the maintenance of spermatogenesis, but the cellular pathways regulating proliferation, differentiation and survival of the undifferentiated spermatogonia are partially known. Glucocorticoid-Induced Leucine Zipper (GILZ), is a mediator of the anti-inflammatory and immunomodulatory effects of glucocorticoids because GILZ controls the cell proliferation and differentiation. L-GILZ, the longer isoform of GILZ, is the only isoform expressed in spermatogonia and primary spermatocytes. The aim of this project is to understand the role of L-GILZ in the control of spermatogenesis. In this work I show that L-GILZ is an essential mediator of the spermatogenesis. Indeed, gilz deficiency in knock-out mice leads to a complete loss of germ cell lineage within first cycles of spermatogenesis, resulting in male sterility with a phenotype similar to pathological features of Sertoli Cell-Only Syndrome. Spermatogenesis failure is intrinsic to germ cells and is associated with increased proliferation and aberrant differentiation of undifferentiated spermatogonia, with hyperactivity of Ras signaling pathway, and with a massive apoptosis during meiotic phase. This discovery is a clear advance in our knowledge of the mechanisms underlying spermatogenesis and indicate L-GILZ as a possible marker for diagnosis and a target for novel therapies to intervene in male sterility.</br

GILZ promotes production of peripherally induced Treg cells and mediates the crosstalk between glucocorticoids and TGF-β signaling

Summary: Regulatory T (Treg) cells expressing the transcription factor forkhead box P3 (FoxP3) control immune responses and prevent autoimmunity. Treatment with glucocorticoids (GCs) has been shown to increase Treg cell frequency, but the mechanisms of their action on Treg cell induction are largely unknown. Here, we report that glucocorticoid-induced leucine zipper (GILZ), a protein induced by GCs, promotes Treg cell production. In mice, GILZ overexpression causes an increase in Treg cell number, whereas GILZ deficiency results in impaired generation of peripheral Treg cells (pTreg), associated with increased spontaneous and experimental intestinal inflammation. Mechanistically, we found that GILZ is required for GCs to cooperate with TGF-β in FoxP3 induction, while it enhances TGF-β signaling by binding to and promoting Smad2 phosphorylation and activation of FoxP3 expression. Thus, our results establish an essential GILZ-mediated link between the anti-inflammatory action of GCs and the regulation of TGF-β-dependent pTreg production. : Peripherally induced Treg cells (pTreg) are generated outside of the thymus and regulate responses to foreign antigens. In this manuscript, Riccardi and colleagues demonstrate that glucocorticoid-induced protein GILZ controls generation of pTreg cells and colon homeostasis. GILZ promotes TGF-β-induced phosphorylation of Smad2 and the expression of FoxP3. Thus, GILZ mediates a synergy between glucocorticoids and TGF-β in pTreg cell induction. GILZ is essential for Treg induction by glucocorticoids and their anti-inflammatory activity

Vitamin D status and the relationship with bone fragility fractures in HIV-infected patients: A case control study

HIV-infected patients show high risk of fracture. The aims of our study were to determine the prevalence of vertebral fractures (VFs) and their associations with vitamin D in HIV patients. 100 patients with HIV infection and 100 healthy age-and sex-matched controls were studied. Bone mineral density was measured by quantitative ultrasound at the non-dominant heel. Serum osteocalcin and C-terminal telopeptide of collagen type 1 served as bone turnover markers. Bone ultrasound measurements were significantly lower in patients compared with controls (Stiffness Index (SI): 80.58 ± 19.95% vs. 93.80 ± 7.10%, respectively, p &lt; 0.001). VFs were found in 16 patients and in 2 controls. HIV patients with vertebral fractures showed lower stiffness index (SI) (70.75 ± 10.63 vs. 83.36 ± 16.19, respectively, p = 0.045) and lower vitamin D levels (16.20 ± 5.62 vs. 28.14 ± 11.94, respectively, p &lt; 0.02). The majority of VFs (87.5%) were observed in HIV-infected patients with vitamin D insufficiency, and regression analysis showed that vitamin D insufficiency was significantly associated with vertebral fractures (OR 9.15, 95% CI 0.18-0.52, p &lt; 0.04). VFs and are a frequent occurrence in HIV-infected patients and may be associated with vitamin D insufficiency

Atypical Presentation of Herpes Simplex Virus 1 Encephalitis in Pediatric Age

BACKGROUND: The annual incidence of herpes simplex encephalitis (HSE) is 1:250–500 thousand in developed countries, which makes it the most common viral cause of encephalitis. Encephalitis caused by HSV-1 probably accounts for most of the cases in the Herpesviridae family and typically affects older children. CASE REPORT: An atypical presentation of pediatric herpetic encephalitis is reported. The child presented with recurrent seizures that were initially unresponsive to several anticonvulsant drugs. EEG and MRI of the brain were not effective for diagnosis. PCR analysis of the cerebrospinal fluid indicated positivity for HSV DNA and was crucial for the diagnosis as a highly sensitive and specific test. CONCLUSION: Based on the clinical presentation of the child, viral encephalitis was deemed to be the most likely aetiological condition

p62/SQSTM1 expression in canine mammary tumours: Evolutionary notes

Abstract Recent studies highlighted the role of autophagy as a cardinal regulatory system for homeostasis and cancer-related signalling pathways. In this context, the deregulated expression of p62 – Sequestosome1 (p62/SQSTM1) – a protein acting both as an autophagy receptor and signalling hub, has been associated with tumour development and chronic inflammation. Multiple clinical studies test drugs targeting autophagy, and even more research is on the way to clinical trials. However, no comparative investigations have been carried out to identify adequate preclinical models to assess p62-based medicine. In veterinary oncology the role of p62 in cancer related pathways has been largely ignored. We compared p62 sequences in multiple organisms and found that canine p62 significantly diverges from the humans and from other animals sequences. Then, we chart by immunohistochemistry the expression levels of p62 in canine mammary tumours. A total of 66 tumours and 10 nonneoplastic mammary samples were examined. The expression of p62 was higher in normal tissue and adenomas than carcinomas, with lowest levels of p62 protein detected in high grade carcinomas. In all cases examined the tumour stroma appeared to be p62-negative. Taken together our results would suggest that in dogs the association between p62 expression and cancer cells overturns that reported in humanbreast carcinoma, where p62 accumulates in malignant cells as compared to normal epithelium. Thus, at least in canine mammary tumours, p62 should be not considered a tumour-rejection antigen for an anti-cancer immunotherapy

Recombinant DNA Technology for Melanoma Immunotherapy: Anti-Id DNA Vaccines Targeting High Molecular Weight Melanoma-Associated Antigen

Anti-idiotypic MK2-23 monoclonal antibody (anti-Id MK2-23 mAb), which mimics the high molecular weight melanoma-associated antigen (HMW-MAA), has been used to implement active immunotherapy against melanoma. However, due to safety and standardization issues, this approach never entered extensive clinical trials. In the present study, we investigated the usage of DNA vaccines as an alternative to MK2-23 mAb immunization. MK2-23 DNA plasmids coding for single chain (scFv) MK2-23 antibody were constructed via the insertion of variable heavy (VH) and light (VL) chains of MK2-23 into the pVAC-1mcs plasmids. Two alternative MK2-23 plasmids format VH/VL, and VL/VH were assembled. We demonstrate that both polypeptides expressed by scFv plasmids in vitro retained the ability to mimic HMW-MAA antigen, and to elicit specific anti-HMW-MAA humoral and cellular immunoresponses in immunized mice. Notably, MK2-23 scFv DNA vaccines impaired the onset and growth of transplantable B16 melanoma cells not engineered to express HMW-MAA. This pilot study suggests that optimized MK2-23 scFv DNA vaccines could potentially provide a safer and cost-effective alternative to anti-Id antibody immunization, for melanoma immunotherapy