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To extend the preservation of donor hearts beyond the current 4-6 h, this paper explores heart cryopreservation by vitrification-cryogenic storage in a glass-like state. While organ vitrification is made possible by using cryoprotective agents (CPA) that inhibit ice during cooling, failure occurs during convective rewarming due to slow and non-uniform rewarming which causes ice crystallization and/or cracking. Here an alternative, "nanowarming", which uses silica-coated iron oxide nanoparticles (sIONPs) perfusion loaded through the vasculature is explored, that allows a radiofrequency coil to rewarm the organ quickly and uniformly to avoid convective failures. Nanowarming has been applied to cells and tissues, and a proof of principle study suggests it is possible in the heart, but proper physical and biological characterization especially in organs is still lacking. Here, using a rat heart model, controlled machine perfusion loading and unloading of CPA and sIONPs, cooling to a vitrified state, and fast and uniform nanowarming without crystallization or cracking is demonstrated. Further, nanowarmed hearts maintain histologic appearance and endothelial integrity superior to convective rewarming and indistinguishable from CPA load/unload control hearts while showing some promising organ-level (electrical) functional activity. This work demonstrates physically successful heart vitrification and nanowarming and that biological outcomes can be expected to improve by reducing or eliminating CPA toxicity during loading and unloading.

Pancreatic islet transplantation can cure diabetes but requires accessible, high-quality islets in sufficient quantities. Cryopreservation could solve islet supply chain challenges by enabling quality-controlled banking and pooling of donor islets. Unfortunately, cryopreservation has not succeeded in this objective, as it must simultaneously provide high recovery, viability, function and scalability. Here, we achieve this goal in mouse, porcine, human and human stem cell (SC)-derived beta cell (SC-beta) islets by comprehensive optimization of cryoprotectant agent (CPA) composition, CPA loading and unloading conditions and methods for vitrification and rewarming (VR). Post-VR islet viability, relative to control, was 90.5% for mouse, 92.1% for SC-beta, 87.2% for porcine and 87.4% for human islets, and it remained unchanged for at least 9 months of cryogenic storage. VR islets had normal macroscopic, microscopic, and ultrastructural morphology. Mitochondrial membrane potential and adenosine triphosphate (ATP) levels were slightly reduced, but all other measures of cellular respiration, including oxygen consumption rate (OCR) to produce ATP, were unchanged. VR islets had normal glucose-stimulated insulin secretion (GSIS) function in vitro and in vivo. Porcine and SC-beta islets made insulin in xenotransplant models, and mouse islets tested in a marginal mass syngeneic transplant model cured diabetes in 92% of recipients within 24–48 h after transplant. Excellent glycemic control was seen for 150 days. Finally, our approach processed 2,500 islets with >95% islets recovery at >89% post-thaw viability and can readily be scaled up for higher throughput. These results suggest that cryopreservation can now be used to supply needed islets for improved transplantation outcomes that cure diabetes.

Partial liver resections are routinely performed in living donor liver transplantation and to debulk tumors in liver malignancies. Due to the complexity of the interconnected hepatic vasculature, pre- and intra-operative decisions on vessel reconstruction for adequate inflow and outflow are challenging. Pre-operative evaluation of hepatic vasculature has predominantly been limited to radiological imaging and has failed to account for post-resection hepatic hemodynamic alterations. Substantial evidence suggests post-surgical increase in local volume flow rate enhances shear stress, signaling hepatic regeneration that is critical for hepatic volume restoration. However, excessive shear stress from extensive hepatic volume reduction has been postulated to result in small for size syndrome and liver failure. Predicting alterations in shear stress, flow velocity, and speed throughout the liver is particularly challenging due to the dendritic architecture of the portal and arterial vasculature that spans several orders of magnitude in diameter. Therefore, we developed a mathematical lumped parameter model implementing realistic heterogeneities in hepatic inflow/outflow of the human liver to simulate surgical resection alterations in perfusion across all hepatic segments. Our model is parameterized based on clinical measurements, relies on only a single free parameter, and accurately captures established blood perfusion characteristics. We quantify changes in volume flow rate, average blood flow velocity, and wall shear stress following variable, realistic liver resections and make comparisons to the intact liver. Our numerical model runs on a laptop in minutes and can be adapted to individualized patient-specific anatomy, providing a substantial advancement towards a novel computational tool for assisting pre- and intra-operative surgical decisions for liver resections.

BACKGROUND Direct Observed Treatment (DOT) is a requirement in the management of Tuberculosis (TB) globally. With the transition from alternate day treatment to daily treatment in India, monitoring treatment adherence through DOT is a logistic challenge. The pervasiveness of mobile phones in India therefore provides a unique opportunity to address this challenge remotely. OBJECTIVE This study was designed to compare the acceptability of mobile phones for antitubercular treatment (ATT) support in two distinct regions of India. METHODS This was a cross sectional exploratory study that enrolled 351 patients with TB, of whom 185 were from Bangalore, Karnataka, South India and 166 from Ujjain, Madhya Pradesh, Central India. Trained research assistants administered a pretested questionnaire comprising demographics, phone usage patterns and acceptability of mobile phone technology to support treatment adherence to TB medicines. The results were statistically analysed. RESULTS The mean age of the 351 participants was 32±13.6 years of whom 140 (40%) were women. Of the participants, 259 (74%) were urban, 221 (63%) had >4 years of education. A significantly greater number of participants were newly diagnosed with TB and were in the intensive phase of treatment. Overall, 218 (62%) preferred vDOT over DOT. There was an overall difference in preference between the two sites which is explained by difference in socio economic variables vDOT CONCLUSIONS Mobile phone adherence support is acceptable to patients on Antitubercular treatment ATT with minor variations in design based on demographic and cultural differences. In India, the preference for voice calls over text messages/SMS while designing mHealth interventions cannot be ignored. Of importance is the preference for DOT over vDOT in central India, unlike South India. However, in time, the expanding use of mobile technology supplemented with counselling, could overcome the barriers of privacy and stigma and promote the transition from in-person DOT to vDOT or mobile phone adherence monitoring and support for ATT in India. CLINICALTRIAL CTRI/2017/07/009052

Pancreatic islet transplantation can cure diabetes but requires accessible, high-quality islets in sufficient quantities. Cryopreservation could solve islet supply chain challenges by enabling quality-controlled banking and pooling of donor islets. Unfortunately, cryopreservation has not succeeded in this objective, as it must simultaneously provide high recovery, viability, function and scalability. Here, we achieve this goal in mouse, porcine, human and human stem cell (SC)-derived beta cell (SC-beta) islets by comprehensive optimization of cryoprotectant agent (CPA) composition, CPA loading and unloading conditions and methods for vitrification and rewarming (VR). Post-VR islet viability, relative to control, was 90.5% for mouse, 92.1% for SC-beta, 87.2% for porcine and 87.4% for human islets, and it remained unchanged for at least 9 months of cryogenic storage. VR islets had normal macroscopic, microscopic, and ultrastructural morphology. Mitochondrial membrane potential and adenosine triphosphate (ATP) levels were slightly reduced, but all other measures of cellular respiration, including oxygen consumption rate (OCR) to produce ATP, were unchanged. VR islets had normal glucose-stimulated insulin secretion (GSIS) function in vitro and in vivo. Porcine and SC-beta islets made insulin in xenotransplant models, and mouse islets tested in a marginal mass syngeneic transplant model cured diabetes in 92% of recipients within 24-48 h after transplant. Excellent glycemic control was seen for 150 days. Finally, our approach processed 2,500 islets with95% islets recovery at89% post-thaw viability and can readily be scaled up for higher throughput. These results suggest that cryopreservation can now be used to supply needed islets for improved transplantation outcomes that cure diabetes.

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.

Coronavirus has emerged as a global health threat due to its accelerated geographic spread over the last two decades. This article reviews the current state of knowledge concerning the origin, transmission, diagnosis and management of coronavirus disease 2019 (COVID-19). Historically, it has caused two pandemics: severe acute respiratory syndrome and Middle East respiratory syndrome followed by the present COVID-19 that emerged from China. The virus is believed to be acquired from zoonotic source and spreads through direct and contact transmission. The symptomatic phase manifests with fever, cough and myalgia to severe respiratory failure. The diagnosis is confirmed using reverse transcriptase PCR. Management of COVID-19 is mainly by supportive therapy along with mechanical ventilation in severe cases. Preventive strategies form the major role in reducing the public spread of virus along with successful disease isolation and community containment. Development of a vaccine to eliminate the virus from the host still remains an ongoing challenge.

Context: Buses take up more than 90% of public transport in Indian cities and serve as a cheap and convenient mode of transport for all classes of society. However, the well-being of employees of this mode of transport is paramount in passenger and personal safety. As in any job, the person has to balance work and personal issues. Work–life balance is a concept that supports the effort from an employee in any sector to split their time and energy to balance work and personal lives. Materials and Methods: A cross-sectional study using a structured interview schedule was conducted among 103 Karnataka State Road Transport Corporation (KSRTC) workers in Anekal town, Karnataka, to determine the work–life balance among them. The total score was classified into high, medium, and low work balance categories. The scores for each domain were analyzed separately to derive at the factors which act against work–life balance. Results: Low work–life balance was found in 26 (25.2%) of all subjects. The important factors that affected work–life balance were identified to be shift work, work load, night duties, social functions, and negative attitude of family members. The reasons that motivated them to work were to support family (35%), future security (33%), and to clear personal debts (15.5%). Conclusion: Given the low work–life balance in a quarter of several individuals in this study, there is a need to address this issue to ensure necessary balance and safety and well-being of both road transport employees and passengers.

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.

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.

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 CD(4) counts, a marker of immunologic status, correlates with FNAC morphology of TBL. This study was undertaken to compare the mean CD(4) counts across the different cytology patterns and to correlate the CD(4) counts with FNAC patterns in these patients.Forty newly diagnosed HIV patients with TBL on FNAC (10 in each pattern) were selected by convenient sampling based on inclusion exclusion criteria. The CD(4) counts were obtained in these patients. Its correlation with different FNAC patterns was assessed using SPSS version 16.Analysis of covariance showed significant difference in the mean CD(4) counts between all the four patterns [F (374), df (3), P-value = 0.000]. Spearman's correlation analysis showed significant correlation of CD(4) counts with the FNAC patterns (correlation coefficient of 0.967; P-value of 0.01) with pattern 1 having low CD(4) counts and pattern 4 having high CD(4) counts.CD(4) 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 CD(4) counts, while pattern 4, suggestive of good immunological response (well defined granuloma, no necrosis and AFB 1+), had high CD(4) counts. Thus FNAC patterns may be used to predict the CD(4) counts in HIV patients where CD(4) facilities are not available or vice versa.