MicroRNAs delivery into human cells grown on 3D-printed PLA scaffolds coated with a novel fluorescent PAMAM dendrimer for biomedical applications

Many advanced synthetic, natural, degradable or non-degradable materials have been employed to create scaffolds for cell culture for biomedical or tissue engineering applications. One of the most versatile material is poly-lactide (PLA), commonly used as 3D printing filament. Manufacturing of multifunctional scaffolds with improved cell growth proliferation and able to deliver oligonucleotides represents an innovative strategy for controlled and localized gene modulation that hold great promise and could increase the number of applications in biomedicine. Here we report for the first time the synthesis of a novel Rhodamine derivative of a poly-amidoamine dendrimer (G = 5) able to transfect cells and to be monitored by confocal microscopy that we also employed to coat a 3D-printed PLA scaffold. The coating do not modify the oligonucleotide binding ability, toxicity or transfection properties of the scaffold that is able to increase cell proliferation and deliver miRNA mimics (i.e., pre-mir-503) into human cells. Although further experiments are required to optimize the dendrimer/miRNA ratio and improve transfection efficiency, we demonstrated the effectiveness of this promising and innovative 3D-printed transfection system to transfer miRNAs into human cells for future biomedical applications. © 2018, The Author(s)

Imaging and Endoscopic Diagnosis of Lung Diseases in Small Animals. A Review

Diagnostic imaging plays a fundamental role in the diagnosis of pulmonary diseases. Radiography, ultra-sound, computed tomography, and endoscopy are important tools for achieving a diagnosis. The choice of diagnostic procedure varies according to the patient, the suspected diagnosis and the risk/benefit ratio. Cul-ture, cytology and histology are nearly always necessary to obtain a definitive diagnosis. Several biopsy sam-pling techniques are described. Surgical biopsies are the gold standard for the diagnosis of bronchiolitis or interstitial lung diseases but often not performed due to the high risk. In humans, the introduction of trans -bronchial cryobiopsies has led to excellent results in the study of interstitial lung diseases. (c) 2022 Elsevier Inc. All rights reserved

A Comparison of Times Taken for the Placement of the First portal and Complication Rates between the Veress Needle Technique and the Modified Hasson Technique in Canine Ovariectomy Laparoscopic Surgery

Minimally invasive surgery is increasingly being used in veterinary medicine. Laparoscopic procedures have several advantages compared with open surgery. These include the magnification of the field of surgery, reduced post-surgical pain and associated stress, reduced post-operative infection rates, and decreased hospitalization time. The establishment of a pneumoperitoneum is a critical step; however, this procedure can prolong the operation time, and most of the complications associated with laparoscopic surgery have been attributed to the insertion of devices into the abdominal cavity. Two main techniques have been employed to create pneumoperitoneum: the closed-entry method using the Veress needle and the open Hasson technique. The first portal is necessary to start insufflation and, subsequently, to realize the operative channel to insert the laparoscopic instruments into the abdomen. Many authors have compared the time necessary to create the first portal using different techniques in human medicine, but studies on this topic in veterinary medicine are lacking. In the veterinary medicine literature, complications associated with the creation of a pneumoperitoneum and the placement of ports include spleen, bowel, or bladder injuries; pneumothorax; and subcutaneous emphysema. The aim of the present study was to compare the times required for the placement of the first portal and the creation of pneumoperitoneum, and the rates of intraoperative complications using the Veress needle technique (VNT) and the open modified Hasson technique (MHT). The sample population comprised 30 female dogs who underwent laparoscopic ovariectomies. The dogs were randomly organized into two groups and two different entry techniques were used: Veress needle (VNT = group A) and the modified Hasson technique (MHT = group B). Complications related to abdominal entry were classified as major, in cases of organ perforation, and minor, in cases of subcutaneous emphysema and gas leakage. The VNT and MHT required 374.0 s and 242.9 s, respectively, for the placement of the first portal and for establishing pneumoperitoneum (p < 0.05). Their major complications rates were 20% and 0%, respectively (p < 0.05). Their minor complications rates were 20% and 35%, respectively (p < 0.05). No surgical procedures required laparotomy. The MHT was associated with a lower major complication rate and required less time to create the first portal, compared with the Veress needle technique

Subtotal Epiglottectomy and Ablation of Unilateral Arytenoid Cartilage as Surgical Treatments for Grade III Laryngeal Collapse in Dogs

Stage III laryngeal collapse is defined as the collapse of the corniculate processes of the arytenoid cartilages and the destructuration of the dorsal portion of the rima glottidis. The primary cause is chronic upper airway obstruction, and the condition is often present in brachycephalic dogs. The treatment is still controversial; the patients are generally treated with a permanent tracheostomy. This article reports the authors’ experience with 16 dogs affected by stage III laryngeal collapse treated with subtotal epiglottectomy and the ablation of unilateral arytenoid cartilage. Before the surgery, all of the dogs underwent an effort test to classify the clinical severity of the disease and an endoscopic examination of the airways to determine the stage of severity of the laryngeal collapse. One month after surgery, the effort test was repeated in order to evaluate the clinical outcome. One year after surgery, the owners of 12 patients rated their dogs as follows: excellent in five cases, good in five cases, and fair in two cases. According to this pilot study, epiglottectomy associated with the photoablation of unilateral arytenoid cartilage increases airway flow, and thus may be considered a valid surgical procedure to treat dogs affected by grade III laryngeal collaps

Two-Port Laparoscopic Adrenalectomy in Dogs

SIMPLE SUMMARY: An adrenal mass represents the primary indication for adrenalectomy. Laparoscopic adrenalectomy is a minimally invasive surgical technique that minimizes pain and improves patient recovery. The traditional technique involves the use of a single or multiple port (three or four). The objective of the study was to evaluate the feasibility and complication rate of two-port laparoscopic adrenalectomy to remove adrenal masses smaller than 4 cm in diameter. In total, 16 dogs with adrenal masses were enrolled in the study and clinicopathologic data were harvested from the medical records. The dogs were placed in sternal recumbency with cushions positioned under the sternum and pubis to simulate the quadrupedal station and displace the abdominal viscera. In all cases, the procedures were feasible with two ports. In five cases, capsule rupture occurred, and all were adrenal gland carcinomas. ABSTRACT: The gold-standard treatment for functional tumors is adrenalectomy, and the procedure can be either open or laparoscopic. Laparoscopic adrenalectomy (LA) is a minimally invasive technique designated for small–medium-sized adrenal tumors without vascular invasion. LA is routinely performed using three or four ports with the patient in sternal or lateral recumbency. The aim of the study was to evaluate the feasibility of LA with two ports in order to reduce invasiveness and improve patient recovery. In total, 16 dogs with adrenal tumors were included in the study and the two-port technique was performed. Adrenalectomy was performed based on the presence of hypercortisolism in thirteen cases, whereas, in three cases, adrenalectomy was performed in the absence of evidence of cortisol production. Thirteen cases were carcinomas and three were adenomas of the adrenal gland; furthermore, twelve were on the left side and four were on the right side. Capsule rupture occurred in five cases. After performing the technique in all cases, no additional ports or laparotomy conversion occurred. Based on the authors’ experience, laparoscopic adrenalectomy can be performed with two ports

Antibodies from patients with rheumatoid arthritis target citrullinated histone 4 contained in neutrophils extracellular traps.

Histone deimination regulates gene function and contributes to antimicrobial response, allowing the formation of neutrophil extracellular traps (NETs). Deiminated proteins are target of anti-citrullinated peptides antibodies (ACPA) in rheumatoid arthritis (RA). OBJECTIVE: The objective of this paper is to test the hypothesis that RA sera react with deiminated histones contained in NETs. METHODS: Neutrophils from peripheral blood were stimulated with A23187 and acid treated; NETosis was induced by phorbol myristate acetate, and NET proteins were isolated. Sera were tested by immunoblot on acid extracted proteins from neutrophils and from NETs, and by ELISA on deiminated histone H4 or H4-derived peptides. Bands reactive with RA sera were excised from gels, digested with trypsin and subjected to matrix-assisted laser desorption/ionisation time of flight (MALDI-TOF) analysis, before and after derivatisation to detect citrullinated peptides. RESULTS: RA sera reacted with a deiminated antigen of 11 KDa from activated neutrophils, recognised also by anti-H4 and antideiminated H4 antibodies. A similar reactivity was observed with NET proteins. The antigen from neutrophils or NETs was identified as citrullinated H4 by MALDI-TOF analysis. By ELISA, RA sera bound in vitro citrullinated H4. Citrullinated H4 14-34 and 31-50 peptides detected antibodies in 67% and 63% of RA sera and in less than 5% of controls; antibody titre was correlated with anti-CCP2. CONCLUSIONS: Citrullinated H4 from activated neutrophils and NETs is a target of antibodies in RA, and synthetic citrullinated H4-derived peptides are a new substrate for ACPA detection. As NETosis can generate antigens for ACPA, these data suggest a novel connection between innate and adaptive immunity in RA

Serological and genetic evidence for altered complement system functionality in systemic lupus erythematosus: Findings of the GAPAID consortium

Systemic lupus erythematosus is a chronic autoimmune disease with multifactorial ethiopathogenesis. The complement system is involved in both the early and late stages of disease development and organ damage. To better understand autoantibody mediated complement consumption we examined ex vivo immune complex formation on autoantigen arrays. We recruited patients with SLE (n = 211), with other systemic autoimmune diseases (n = 65) and non-autoimmune control subjects (n = 149). Standard clinical and laboratory data were collected and serum complement levels were determined. The genotype of SNP rs1143679 in the ITGAM gene was also determined. Ex vivo formation of immune complexes, with respect to IgM, IgG, complement C4 and C3 binding, was examined using a functional immunoassay on autoantigen microarray comprising nucleic acids, proteins and lipids. Complement consumption of nucleic acids increased upon binding of IgM and IgG even when serum complement levels were decreased due to consumption in SLE patients. A negative correlation between serum complement levels and ex vivo complement deposition on nucleic acid autoantigens is demonstrated. On the contrary, complement deposition on tested protein and lipid autoantigens showed positive correlation with C4 levels. Genetic analysis revealed that the non-synonymous variant rs1143679 in complement receptor type 3 is associated with an increased production of anti-dsDNA IgG antibodies. Notwithstanding, homozygous carriers of the previously reported susceptible allele (AA) had lower levels of dsDNA specific IgM among SLE patients. Both the non-synonymous variant rs1143679 and the high ratio of nucleic acid specific IgG/IgM were associated with multiple organ involvement. In summary, secondary complement deficiency in SLE does not impair opsonization of nucleic-acid-containing autoantigens but does affect other antigens and potentially other complement dependent processes. Dysfunction of the receptor recognizing complement opsonized immune complexes promotes the development of classswitched autoantibodies targeting nucleic acids

Use of a high sensitive nanofluidic array for the detection of rare copies of BCR-ABL1 transcript in patients with Philadelphia-positiveacute lymphoblastic leukemia in complete responseIlaria

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