Identification of an antigenic domain near the C terminus of human granulocyte-macrophage colony-stimulating factor and its spatial localization.

The goal of this study was to map an epitope on the human granulocyte-macrophage colony-stimulating factor (hGM-CSF) at its C terminus, a region whose integrity is fundamental in maintaining the normal function of this molecule. Residues including the fourth alpha-helix (D, 103-116) were analyzed for their role in the interaction with antibodies (Abs) raised against the protein. Five peptides homologous to different segments of the C terminus of hGM-CSF were synthesized. Peptide-(102-121) included the same residues of the alpha-helix D and the next five amino acids toward the C terminus; peptide-[E108A]-(102-121) introduced the mutation E108A in order to verify the role of acidic residues; peptide-[C96A](93-110) encompassed the beta-sheet 2 and half of the alpha-helix D; peptide-[C121A]-(110-127) included the second half of the alpha-helix D and the C terminus of hGMCSF; peptide-(13-31)-Gly-Pro-Gly-(103-116) included both the alpha-helices A and D connected by the tripeptide Gly-Pro-Gly, which allows the original antiparallel orientation of the two alpha-helices to be maintained. Both anti-protein and anti-peptide-(102-121) antibodies, capable of neutralizing the stimulatory activity of hGMCSF in the bone marrow colony-forming assays, recognized a specific epitope in the C terminus of hGM-CSF. Molecular modeling estimated the surface accessibility of hGM-CSF and the stability of the synthetic peptides in aqueous solution. Altogether, our results showed that the immunogenic region includes part of the alpha-helix D and the residues 116-120, which are external to this helix and particularly exposed on the protein surface, confirming the feasible participation of this region in antibody binding

Bilateral reconstruction of the mandibular body with symphyseal preservation using a single fibula free flap: operative technique

Background: Mandibular osteonecrosis may occur in 5% of the patients who undergo radiotherapy for the treatment of head and neck malignancies. Resection and microvascular reconstruction is the treatment of choice in complicated osteoradionecrosis, however multifocal presentation may complicate the management of the disease given the poor quality and limited availability of adequate recipient vessels. Operative technique: A 74-year-old man affected by multifocal severe osteoradionecrosis of the mandible underwent bilateral resection of the mandibular bodies while preserving the symphysis. The defects were reconstructed with a single fibula flap composed by two bony segments connected by a central segment, corresponding to the symphyseal region, in which the bone was dissected and removed. The anastomosis was performed on a single side of the neck. Healing was uneventful and the adopted technique allowed for a quick functional and esthetic recovery. Conclusion: The presented technique provided a safe and efficacious, although technically challenging, solution in a case presenting multifocal osteonecrosis of the jaw. The morbidity of the procedure was limited because the tissue resection and reconstruction processes were minimized. Graphical Abstract: [Figure not available: see fulltext.

Folded Radial Forearm Free Flap for the Reconstruction of Total Soft Palate Defects: Operative Technique

Background: The surgical plan to reconstruct the palate must be carefully prepared given the morphological peculiarity of the soft palate forming both the roof of the mouth and the floor of the nasal cavity. This article focuses on the use of folded radial forearm free flaps to manage isolated defects of the soft palate in the absence of tonsillar pillar involvement. Methods: Three patients affected by squamous cell carcinoma of the palate underwent resection of the soft palate and immediate reconstruction with a folded radial forearm free flap. Results: All three patients showed good short-term morphological-functional outcomes as far as swallowing, breathing, and phonation were concerned. Conclusions: The folded radial forearm free flap seems to be an efficacious way to manage localized defects of the soft palate, given the positive outcomes of the three patients treated, and in accordance with other authors. In general, the radial forearm free flap was confirmed to be a versatile solution for those intraoral defects of the soft tissue requiring a limited quantity of volume as in the case of the soft palate

Ameloblastic fibroma in a 6-year old child:case report.

Ameloblastic fibroma (AF) is defined in WHO classification as a ''neoplasm composed of proliferating odontogenic epithelium embedded in a cellular ectomesenchymal tissue that resembles dental papilla, and with varying degrees of inductive change and dental hard tissue formation''. AF is a rather uncommon tumor, accounting for only 2.5% of all odontogenic tumors. AF is a true mixed tumor, in which the epithelial and ectomesenchymal elements are neoplastic. AF raises at any age, ranging from 6 months to 42 years (mean 14.6 to 15.5 years); it does not show sex predilection. The lesion occurs in nearly 70% of cases in posterior areas of the mandible. Patients exhibit swelling of the jaw; pain is not usually described. Authors present a clinical and surgical management of an early onset of a large mandibular ameloblastic fibroma in a 6-year-old girsl

Adult Human Vascular Smooth Muscle Cells on 3D Silk Fibroin Nonwovens Release Exosomes Enriched in Angiogenic and Growth-Promoting Factors

Background. Our earlier works showed the quick vascularization of mouse skin grafted Bombyx mori 3D silk fibroin nonwoven scaffolds (3D-SFnws) and the release of exosomes enriched in angiogenic/growth factors (AGFs) from in vitro 3D-SFnws-stuck human dermal fibroblasts (HDFs). Here, we explored whether coronary artery adult human smooth muscle cells (AHSMCs) also release AGFs-enriched exosomes when cultured on 3D-SFnws in vitro. Methods. Media with exosome-depleted FBS served for AHSMCs and human endothelial cells (HECs) cultures on 3D-SFnws or polystyrene. Biochemical methods and double-antibody arrays assessed cell growth, metabolism, and intracellular TGF-β and NF-κB signalling pathways activation. AGFs conveyed by CD9+/CD81+ exosomes released from AHSMCs were double-antibody array analysed and their angiogenic power evaluated on HECs in vitro. Results. AHSMCs grew and consumed D-glucose more intensely and showed a stronger phosphorylation/activation of TAK-1, SMAD-1/-2/-4/-5, ATF-2, c-JUN, ATM, CREB, and an IκBα phosphorylation/inactivation on SFnws vs. polystyrene, consistent overall with a proliferative/secretory phenotype. SFnws-stuck AHSMCs also released exosomes richer in IL-1α/-2/-4/-6/-8; bFGF; GM-CSF; and GRO-α/-β/-γ, which strongly stimulated HECs’ growth, migration, and tubes/nodes assembly in vitro. Conclusions. Altogether, the intensified AGFs exosomal release from 3D-SFnws-attached AHSMCs and HDFs could advance grafts’ colonization, vascularization, and take in vivo—noteworthy assets for prospective clinical applications

Reduced T-cell repertoire restrictions in abatacept-treated rheumatoid arthritis patients.

BACKGROUND: CD28(neg) T cells, which display functional characteristic of oligoclonally expanded cytotoxic memory T lymphocytes, are believed to be pathologically relevant in rheumatoid arthritis manifestation. The CD28 co-stimulation blockade by abatacept can prevent the generation of CD28(neg) T-cell populations in these patients. METHODS: Samples were obtained before and after 12 months of abatacept therapy. T-cell phenotype and T-cell receptor diversity were evaluated by flow cytometry and complementarity-determining region-3 spectratyping, respectively, while telomerase reverse-transcriptase gene level was measured by real-time PCR. RESULTS: Abatacept induces a decrease of the percentage and number of CD4(+)CD28(neg) T cells and a reduction of T-cell repertoire restrictions; these features are directly correlated. Thymic output and telomerase activity are not modified by the therapy. CONCLUSIONS: Abatacept-induced decrease of peripheral T-cell repertoire restrictions can due to a reduced generation of senescent, chronically stimulated CD4(+)CD28(neg) T cells

Cryopreservation of pheasant semen: effect of dilution ratio and cooling time on spermatozoa viability and mobility

Aim of the present study was to investigate the cryopreservation of pheasant semen by adopting the freezing/thawing protocol by Tselutin et al. (1999) with some modifications. Semen was collected by abdominal massage twice a week. Evaluations were performed on pooled semen from fifteen males (Phasianus colchicus mongolicus). Two semen dilutions (DIL) with Lake diluent (1:2 and 1:3, v/v; Lake, 1968) and two equilibration times (ET) at 5°C (10min and 30min) before dimethylacetamide (DMA) addition, were tested. Assessment of sperm mobility was performed by Accudenz methodology according to Froman’s procedure (1997) and viability by eosin/nigrosin staining. As expected, viability and mobility were strongly affected by the freezing/thawing procedure. ET did not affect mobility while influenced live sperm percentage during the DMA equilibration (DMA-Eq). Semen/diluent ratio significantly (p<0.001) modified the mobility parameter and the highest progressive movements of spermatozoa were obtained with the most diluted semen in each critical step of the cryopreservation procedure. In conclusion, for pheasant semen cryopreservation, the 1:3 dilution ratio can be considered appropriate and the cooling time up to 30 minutes not crucial for the spermatozoa mobility and viability. Nevertheless, the deleterious effect of freezing/thawing procedure reduced live thawed spermatozoa to 24% and forward motility to 89% of the initial movement capacity

A magnetization transfer study of mild and advanced Parkinson's disease

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