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Background and Aims: Postoperative pulmonary complications (PPCs) lead to increased morbidity, mortality, length of hospital stay, and cost to the patient. This study was conducted to determine the risk factors and assess the incidence of PPC after non-cardiac surgery. Material and Methods: This prospective, observational study was conducted on 1,170 patients undergoing non-cardiac surgery. Details of patient, surgical, and anesthetic factors were collected and patients were followed up for the entire duration of hospital stay for the occurrence of PPC. Assess Respiratory Risk in Surgical Patients in Catalonia (ARISCAT) score and the length of hospital stay was noted for all the patients. Regression analysis was used to find the risk factors associated with development of respiratory complications. Results: The incidence of PPC was found to be 59 in 1,170 patients (5%) in our hospital. Multivariate analysis revealed that patients with intermediate and high risk ARISCAT scoring had higher odds of developing PPC. Higher age (>50 years), positive cough test, presence of nasogastric tube, and intraoperative pulmonary complications were identified as independent risk factors associated with the occurrence of PPC. Conclusion: We found 5% incidence of PPC in our study. Recognition of the delineated risk factors and routine use of ARISCAT score for preoperative assessment may help identify patients at a higher risk of developing postoperative pulmonary complications.

The human leukocyte antigen G1 (HLA-G1), a non-classical class I major histocompatibility complex (MHC-I) protein, is a potent immunomodulatory molecule at the maternal/fetal interface and other environments to regulate the cellular immune response. We created GGTA1-/HLAG1+pigs to explore their use as organ and cell donors that may extend xenograft survival and function in both preclinical nonhuman primate (NHP) models and future clinical trials. In the present study, HLA-G1 was expressed from the porcine ROSA26 locus by homology directed repair (HDR) mediated knock-in (KI) with simultaneous deletion of α-1-3-galactotransferase gene (GGTA1; GTKO) using the clustered regularly interspersed palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9) (CRISPR/Cas9) gene-editing system. GTKO/HLAG1+pigs showing immune inhibitory functions were generated through somatic cell nuclear transfer (SCNT). The presence of HLA-G1 at the ROSA26 locus and the deletion of GGTA1 were confirmed by next generation sequencing (NGS) and Sanger’s sequencing. Fibroblasts from piglets, biopsies from transplantable organs, and islets were positive for HLA-G1 expression by confocal microscopy, flow cytometry, or q-PCR. The expression of cell surface HLA-G1 molecule associated with endogenous β2-microglobulin (β2m) was confirmed by staining genetically engineered cells with fluorescently labeled recombinant ILT2 protein. Fibroblasts obtained from GTKO/HLAG1+pigs were shown to modulate the immune response by lowering IFN-γ production by T cells and proliferation of CD4+and CD8+T cells, B cells and natural killer (NK) cells, as well as by augmenting phosphorylation of Src homology region 2 domain-containing phosphatase-2 (SHP-2), which plays a central role in immune suppression. Islets isolated from GTKO/HLA-G1+genetically engineered pigs and transplanted into streptozotocin-diabetic nude mice restored normoglycemia, suggesting that the expression of HLA-G1 did not interfere with their ability to reverse diabetes. The findings presented here suggest that the HLA-G1+transgene can be stably expressed from the ROSA26 locus of non-fetal maternal tissue at the cell surface. By providing an immunomodulatory signal, expression of HLA-G1+may extend survival of porcine pancreatic islet and organ xenografts.

Vitrification can dramatically increase the storage of viable biomaterials in the cryogenic state for years. Unfortunately, vitrified systems ≥3 mL like large tissues and organs, cannot currently be rewarmed sufficiently rapidly or uniformly by convective approaches to avoid ice crystallization or cracking failures. A new volumetric rewarming technology entitled “nanowarming” addresses this problem by using radiofrequency excited iron oxide nanoparticles to rewarm vitrified systems rapidly and uniformly. Here, for the first time, successful recovery of a rat kidney from the vitrified state using nanowarming, is shown. First, kidneys are perfused via the renal artery with a cryoprotective cocktail (CPA) and silica‐coated iron oxide nanoparticles (sIONPs). After cooling at −40 °C min−1 in a controlled rate freezer, microcomputed tomography (µCT) imaging is used to verify the distribution of the sIONPs and the vitrified state of the kidneys. By applying a radiofrequency field to excite the distributed sIONPs, the vitrified kidneys are nanowarmed at a mean rate of 63.7 °C min−1. Experiments and modeling show the avoidance of both ice crystallization and cracking during these processes. Histology and confocal imaging show that nanowarmed kidneys are dramatically better than convective rewarming controls. This work suggests that kidney nanowarming holds tremendous promise for transplantation., Vitrification dramatically increases organ storage times in the cryogenic state beyond current storage limits. However, vitrified kidneys cannot currently be rewarmed sufficiently rapidly or uniformly to avoid ice crystallization or cracking failures. A new radiofrequency rewarming technology called “nanowarming” addresses this problem by exciting magnetic nanoparticles to rewarm vitrified kidneys rapidly and uniformly.

Introduction Macrophages contribute to xenograft rejection by direct cytotoxicity and by amplifying T cell-mediated immune responses. It has been shown that transgenic expression of hCD47 protects porcine cells from human macrophages by restoring the CD47-SIRPα self-recognition signal. It has also been reported that the long 3' untranslated region (3'UTR) of the hCD47 gene, which is missing from constructs previously used to make hCD47 transgenic pigs, is critical for efficient cell surface expression in human cells. The aim of this study was to investigate the impact of a modified form of the 3'UTR on the expression, localization, and function of hCD47 in transfected porcine cells. Methods hCD47 constructs with and without the modified 3'UTR were knocked into the GGTA1 locus in porcine fetal fibroblasts using CRISPR. Flow cytometry of the transfected cells was used to analyze hCD47 localization. Endoplasmic reticulum (ER), mitochondrial, and oxidative stress were examined by gene expression analysis and confocal microscopy. Phagocytosis of transfected cells by human macrophages was measured by flow cytometry, and stimulation of human/non-human (NHP) primate lymphocytes by the cells was examined using a PBMCs proliferation assay. Results Cells transfected with the construct lacking the 3'UTR (hCD47(3'UTR-)) exhibited predominantly intracellular expression of hCD47, and showed evidence of ER stress, dysregulated mitochondrial biogenesis, oxidative stress, and autophagy. Inclusion of the 3'UTR (hCD47(3'UTR+)) decreased intracellular expression of hCD47 by 36% and increased cell surface expression by 53%. This was associated with a significant reduction in cellular stress markers and a higher level of protection from phagocytosis by human macrophages. Furthermore, hCD47(3'UTR+) porcine cells stimulated significantly less proliferation of human/NHP T cells than hCD47(3'UTR-) cells. Conclusion Our results suggest the potential benefits of using hCD47 constructs containing the 3'UTR to generate genetically engineered hCD47-expressing donor pigs.

Neutrophil–Lymphocyte Ratio (NLR) provides an understanding of the systemic inflammatory conditions. NLR plays an important role as a predictor of mortality in breast and other malignancies. The application of NLR to predict prognosis of Locally Advanced Breast Cancer (LABC) has not been well developed. In this retrospective study, we establish a relationship of pre-treatment NLR with the Pathological Complete Response (pCR) in LABC patients to enhance decision-making and treatment protocols. Data of women diagnosed with carcinoma breast between January 2015 and December 2017 was retrieved from hospital records of a tertiary medical centre in Bangalore, India, after obtaining institutional ethical clearance. LABC patients were categorized into pCR(+) and pCR(−). NLR was calculated and divided into quartiles. The cutoff NLR was determined using the Receiver Operating Characteristic (ROC) curve. Statistical analysis was performed on 119 LABC patients, of which 25 (21%) achieved pCR. Oestrogen Receptor (ER) positivity was significantly lower in pCR(+) than in pCR(−) (p = 0.012). NLR of 2.46 (AUC, 0.744; 95% CI [0.201–0.584]; p = 0.056) was considered the optimum cutoff for pCR(+). A sensitivity of 54%, specificity of 8%, positive predictive value of 1% and high Negative Predictive Value (NPV) of 84% was achieved in the study. A relationship between pCR and the pre-treatment NLR determined a significantly high NPV. Poor pCR in luminal A/B subtype presents with elevated NLR. Therefore, in luminal type A/B (ER- and PR-positive) with elevated NLR (poor outcome) and low pCR (poor response to NACT), the decision of eliminating NACT could be considered, thereby recommending surgical intervention.

Background Fine needle aspiration cytology (FNAC) of TB lymphadenitis (TBL) in HIV shows four different patterns, which may be reflective of immune status. We hypothesize that the CD4 counts, a marker of immunologic status, correlates with FNAC morphology of TBL. This study was undertaken to compare the mean CD4 counts across the different cytology patterns and to correlate the CD4 counts with FNAC patterns in these patients. Methods Forty newly diagnosed HIV patients with TBL on FNAC (10 in each pattern) were selected by convenient sampling based on inclusion exclusion criteria. The CD4 counts were obtained in these patients. Its correlation with different FNAC patterns was assessed using SPSS version 16. Results Analysis of covariance showed significant difference in the mean CD4 counts between all the four patterns [F (374), df (3), P-value = 0.000]. Spearman's correlation analysis showed significant correlation of CD4 counts with the FNAC patterns (correlation coefficient of 0.967; P-value of 0.01) with pattern 1 having low CD4 counts and pattern 4 having high CD4 counts. Conclusion CD4 counts show significant correlation with FNAC patterns of TBL in HIV patients. Pattern 1, suggestive of poor immunological response (chiefly necrosis, occasional ill defined granuloma, AFB 3+) had low CD4 counts, while pattern 4, suggestive of good immunological response (well defined granuloma, no necrosis and AFB 1+), had high CD4 counts. Thus FNAC patterns may be used to predict the CD4 counts in HIV patients where CD4 facilities are not available or vice versa. Diagn. Cytopathol. 2015;43:16–20. © 2014 Wiley Periodicals, Inc.

Background: Systemic Lupus Erythematosus (SLE), a systemic autoimmune disease, is diagnosed by correlating clinical features with a positive Antinuclear Antibody (ANA) test. Detection of ANA by Indirect Immunofluorescence (IIF) is used for screening SLE and is dependent on initial dilution titers which require population-specific standardization. With lack of exclusive commercial kits for the South Indian population, it is necessary to standardize initial screening dilution titers for ANA to distinguish SLE and other rheumatic diseases from the healthy. Methods: Newly diagnosed SLE patients between 18 and 60 years along with healthy controls from South India over 8 months (September 2015–April 2016) were selected for this prospective study. Serum samples were subjected to ANA-IIF in dilution titers of 1:40, 1:80, and 1:100 as per kit recommendations. IIF intensity and staining patterns were correlated clinically and statistically analyzed. Results: ANA positivity in dilutions of 1:40 and 1:80 was seen in 2.1% of healthy controls and negative at 1:100. About 97.9% of SLE patients were positive at 1:100 dilution; speckled pattern being the most common (52.1%), followed by homogeneous (37.4%). The patterns were best appreciated in 1:100 dilution with high significant measure of agreement of kappa between the two pathologists for both patterns and intensity at all three dilutions. Conclusion: 1:100 is the best screening dilution to distinguish SLE patterns from normal healthy individuals, and its main advantage is the delineation of various ANA patterns when positive, especially when mixed at this lower dilution.

Due to the lack of awareness and poor access to laboratory diagnosis, hemophilia may not be diagnosed preoperatively leading to therapeutic misadventure during surgery. Hence, this congenital bleeding disorder due to Factor VIII deficiency reduces surgical management. We report a 39-year-old gentleman, diagnosed of Factor VIII deficiency who presented to emergency with acute spontaneous left hemothorax and underwent a successful thoracotomy and decortication which saved his life. He is positive for human immunodeficiency virus as well as hepatitis B for which he is on treatment. The management guidelines for thoracic surgery are not addressed to in the World Federation of Hemophilia guidelines, making the management challenging in this scenario. We report this case due to its rarity and emphasize that early recognition with immediate surgical intervention supported with Factor VIII transfusion played an important role in saving life.