Non-genetic effects and repeatability estimates of semen production traits in Murrah buffalo bulls.

The objectives of this study were to determine the repeatability estimates and the effect of ejaculate, season, period, age, and bull on semen production traits in Murrah buffalo bulls maintained in an organized semen station. A data set comprising 54,268 ejaculates from 76 Murrah buffalo bulls collected across 12 years were analyzed. The analysis of variables included non-genetic factors as the fixed effects and bull as the random effect. The repeatability was estimated by the restricted maximum likelihood method (REML) using WOMBAT program. The overall least-squares means for semen volume, sperm concentration, total sperm/ejaculate, mass activity, individual motility, post-thaw motility, and frozen semen straws/ejaculate were 2.65 ml, 1222.04 million/ml, 3030.10 million, 2.64, 67.45%, 51.73%, and 128.80 doses. The repeatability estimates were 0.27, 0.25, 0.22, 0.28, 0.34, 0.27, and 0.23, respectively. The studied non-genetic factors influenced (P < 0.01) all seminal traits. Likewise, variation (P < 0.01) due to individual bull was also observed. The first ejaculate was superior in terms of ejaculate volume, sperm concentration, total sperm/ejaculate, and frozen semen straws than the second ejaculate. The south-west monsoon and summer seasons were the best seasons with better semen quality and frozen semen production. Compared to the remaining periods, during Period V (2014-2016), greater values for most seminal traits were found. Bulls above 8 years of age expressed higher semen production than the younger bulls below 4 years, whereas the best performance was observed in the age group of 12 years and above. The repeatability estimates were low to moderate ranging from 0.22 for total sperm/ejaculate to 0.34 for individual motility. A significant (P < 0.01) positive correlation was observed between the seminal traits except the negative correlation in semen volume and total sperm/ejaculate. It was concluded that all the non-genetic factors considered in this study exerted influence on studied seminal traits. Therefore, appropriate management practices can be applied to improve semen quality and quantity.

Assessment of consistency of minimum post-mortem intervals estimated by thermal summation-based methods in medico-legal cases associated with blowflies.

BACKGROUND: This study is based on blow fly samples collected from 8 medico-legal cases in Tamil Nadu, India. The fly life stages were identified and the consistency of minimum post-mortem intervals (PMImin) estimated by different thermal summation-based methods was assessed. METHODS: PMImin of 8 medico-legal cases was estimated using six different thermal summation constants and lower developmental temperatures that are based on C. megacephala and C. rufifacies developmental data. Limits of agreement (LoA), intra class correlation coefficient (ICC) between PMImin values and margin of error of mean of difference between PMImin values were calculated. RESULTS: Intra-class correlation between the PMImin values estimated using different thermal summation constants based on C. megacephala ranged between 0.89 and 0.98 and coefficient of determination ranged between 0.93 and 0.98. Intra-class correlation between the PMImin values estimated using different thermal summation constants based on C. rufifacies ranged between 0.91 and 0.99 and coefficient of determination ranged between 0.95 and 0.99. The mean difference of PMImin values estimated using different thermal summation methods based on C. megacephala ranged between 1.8 hr and 6.6hr and margin of error ranged between 2.51 and 6.93hr. The mean difference of PMImin values estimated using different thermal summation methods based on C. rufifacies ranged between 3.33 and 31.33hr and margin of error ranged between 4.66 and 32hr. CONCLUSION: Consistency of PMImin values estimated by different thermal summation methods was good to excellent. Thermal summation constants useful in estimation of PMImin with lowest mean difference and margin of error were described.

Integrated proteomics and phosphoproteomics revealed druggable kinases in neoadjuvant chemotherapy resistant tongue cancer.

Tongue squamous cell carcinoma is an aggressive oral cancer with a high incidence of metastasis and poor prognosis. Most of the oral cavity cancer patients present in clinics with locally advanced unresectable tumors. Neoadjuvant treatment is beneficial for these individuals as it reduces the tumor size aiding complete resection. However, patients develop therapy resistance to the drug regimen. In this study, we explored the differential expression of proteins and altered phosphorylation in the neoadjuvant chemotherapy resistant tongue cancer patients. We integrated the proteomic and phosphoproteomic profiles of resistant (n = 4) and sensitive cohorts (n = 4) and demonstrated the differential expression and phosphorylation of proteins in the primary tissue of the respective subject groups. We observed differential and extensive phosphorylation of keratins such as KRT10 and KRT1 between the two cohorts. Furthermore, our study revealed a kinase signature associated with neoadjuvant chemotherapy resistance. Kinases such as MAPK1, AKT1, and MAPK3 are predicted to regulate the resistance in non-responders. Pathway analysis showed enrichment of Rho GTPase signaling and hyperphosphosphorylation of proteins involved in cell motility, invasion, and drug resistance. Targeting the kinases could help with the clinical management of neoadjuvant chemotherapy-resistant tongue cancer.

Genetic evaluation of semen production potential in Indigenous bulls using random regression models in a Bayesian framework.

Genetic potential of indigenous bulls with respect to semen production traits over the age of the bulls at semen collection was analyzed using random regression models (RRMs). Data pertaining to 59,641 ejaculates from 189 bulls of 18 indigenous breeds collected from BAIF (Bharatiya Agro Industries Foundation) was utilized for this study. Six semen production traits, viz., ejaculate volume (EV, ml), sperm concentration (SC, 109/ml), initial sperm motility (ISM, %), post-thaw motility (PTM, %), the total number of spermatozoa per ejaculate (TNS, 109/ejaculate), and the theoretical number of semen doses (TNSD) were studied. Univariate and RRM were used to obtain variance components and genetic parameter estimates. Two hundred thousand Gibbs samples were generated for each trait with a burn-in of 20,000 and thinning interval of 50 in a Bayesian framework. Legendre polynomials with orders of fit up to 5 for additive and permanent environmental effects were used. RRM modeled the heritability and repeatability for all ages between 3 and 10 years (productive lifespan). Heritability estimates ranged from 0.18 to 0.36, 0.18 to 0.45, 0.02 to 0.06, 0 to 0.001, 0.09 to 0.32, and 0.14 to 0.42 while the repeatability estimates ranged from 0.41 to 0.72, 0.36 to 0.79, 0.04 to 0.10, 0 to 0.001, 0.37 to 0.56, and 0.32 to 0.57 for EV, SC, ISM, PTM, TNS, and TNSD, respectively. Variability of estimates over the age of the bulls obtained through RRM could be useful to further refine the breeding program for age at selection, deciding the production period and age at culling.

Milk yield, composition, and lactation curve models of Bargur cattle under farmers' production system in their native habitat.

Milk yield and composition traits (fat (%) and SNF (%)) from 1229 test day records of 205 Bargur cattle maintained under a farmers' production system were studied. This breed is known for its adaptability to the hilly tracts of Erode District in Tamil Nadu. Performance recording was done in the natural habitat through the test-day milk yield from lactating cows in the farmers' herds. Effects of non-genetic factors like season, year, parity, stage of lactation, and days from calving were studied using mixed models with animal taken as a random effect. Estimated marginal means for daily milk yield (DMY, L/day), fat (%), and SNF (%) were 2.05 ± 0.03, 4.08 ± 0.03%, and 8.19 ± 0.01%, respectively. Stage of lactation was highly significant (P < 0.01) for DMY, fat, and SNF. Season and year were significant for DMY and SNF, where higher productivity was obtained in the monsoon season, but fat remained constant in all the seasons. Parity was significant (P < 0.05) only for SNF, and year was significant for DMY (P < 0.01) and SNF (P < 0.05). Persistency was 5.9% calculated using Wood's gamma function and 63.0% calculated using the ratio of predicted yields. This was slightly lower compared to dairy breeds of cattle. Thirteen different lactation curve models were used for fitting of Bargur cattle lactation curve, and the Parabolic exponential model was the best fitting model based on model diagnostic criteria.

Performance characteristics of virus neutralization test (VNT) and liquid phase blocking ELISA (LPBE) and their relationship in the cattle immunized with trivalent foot and mouth disease vaccine.

Virus neutralization test (VNT) and liquid phase blocking ELISA (LPBE) are accepted tests for screening and as in vitro alternativ to challenge in FMD vaccine potency testing. To replace VNT by LPBE for the screening of cattle, the optimized tests need to be first evaluated for their diagnostic performances. To replace it with LPBE in the absence of protection data, the interrelationship between VNT and LPBE have to be established to find out LPBE cut­off titer corresponding to the currently used VNT titers. Accordingly, VNT and LPBE were carried out using known negative (n = 306) and positive samples [Serotype O (n = 43), A (n = 14) and Asia1 (n = 11)], for the initial screening. The cut­off of < 1.5 log10 LPBE was comparable with that of < 1.2 log10 VNT titer for screening. LPBE was comparable to VNT in terms of specificity, sensitivity as shown by ROC curve and least varying (coefficient of variation 7.73% in LPBE vs 24.19% in VNT). Based on linear regression model using 471 bovine sera, the predicted LPBE titers corresponding to the currently used log 10 VNT titers of 1.65, 1.5 and 1.5, were 2.24, 1.87 and 2.00 for O, A and Asia1, respectively. These LPBE titers hence can be used as cut­off titers for classifying cattle as protected or not protected until correlation based on in vivo challenge between protection and antibody titer is established.

Precision-weighted estimates of neonatal, post-neonatal and child mortality for 640 districts in India, National Family Health Survey 2016.

BACKGROUND: The conventional indicators of infant and under-five mortality are aggregate deaths occurring in the first year and the first five years, respectively. Monitoring deaths by <1 month (neonatal), 1-11 months (post-neonatal), and 12-59 months (child) can be more informative given various etiological causes that may require different interventions across these three mutually exclusive periods. For optimal resource allocation, it is also necessary to track progress in robust estimates of child survival at a smaller geographic and administrative level. METHODS: Data on 259 627 children came from the 2015-2016 Indian National Family Health Survey. We used a random effects model to account for the complex survey design and sampling variability, and predicted district-specific probabilities of neonatal, post-neonatal, and child mortality. The resulting precision-weighted estimates are more reliable as they pool information and borrow strength from other districts that share the same state membership. The Pearson correlation and Spearman's rank correlation were assessed for the three mortality estimates, and the Moran's I measure was used to detect spatial clustering of high burden districts for each outcome. RESULTS: The majority of under-five deaths was disproportionately concentrated in the neonatal period. Across all districts, the predicted probability of neonatal, post-neonatal, and child mortality varied from 6.0 to 63.9 deaths, 3.8 to 47.6 deaths, and 1.7 to 11.8 deaths per 1000 live births, respectively. The overall correlation between district-wide probabilities of mortality for the three mutually exclusive periods was moderate (Pearson correlation = 0.47-0.58, Spearman's rank correlation = 0.58-0.64). For each outcome, a relatively strong spatial clustering was detected across districts that transcended state boundaries (Moran's I = 0.61-0.76). CONCLUSIONS: Sufficiently breaking down the under-five mortality to distinct age groups and using the precision-weighted estimations to monitor performances at smaller geographic and administrative units can inform more targeted interventions and foster accountability to improve child survival.

Genetic and non-genetic factors affecting semen production and quality characteristics of Gir cattle breed under semi-arid climate.

AIM: This study aims to evaluate genetic and non-genetic factors influencing semen production potential of Gir bulls. MATERIALS AND METHODS: Data on semen quantity (n=6911) and quality (n=466) available from January 2011 to December 2018 at BAIF's frozen semen station, Jind, Haryana, India, were utilized for the study. Factors, namely, season of collection, age at collection, and bull (random effect) were studied for their effect on quantitative and qualitative semen traits. Least square means for the traits were obtained using a general linear model. The effect of age within bull for repeatable traits was analyzed using a longitudinal model with age as the control variable. Multivariate analysis using mixed repeatability model equation was utilized to estimate bull effect correlation (genetic + permanent environmental correlation), phenotypic correlations, and repeatability. RESULTS: The overall least square means of ejaculate volume, sperm concentration, total sperms, initial and post-thaw motility, hypo-osmotic swelling test, and acrosome integrity of frozen semen were 6.62±0.03 ml, 1.22±0.01 109/ml, 8.09±0.05 109/ejaculate, 75.78±0.001%, 55.92±0.0001%, 55.13±0.005%, and 71.08±0.001%, respectively. The season of the collection showed a significant effect on volume, concentration, total sperm, and initial motility. The performance of bulls was superior in summer season, followed by winter and monsoon. Increase in semen attributes during summer season was due to the effect of lower temperature on sensitive stages of spermatogenesis. Age at collection had a significant effect on all semen traits. Volume and total sperm count showed increasing trend while concentration showed a decreasing trend with an increase in age. Motility and quality traits did not show any particular pattern. Individual bulls showed differences in all the semen performance traits with age. The repeatability of the traits ranged from 0.04 (HOST) to 0.58 (acrosome integrity). Bull effect correlation ranged from -0.73 (initial motility and acrosome integrity) to 0.93 (HOST and acrosome integrity). CONCLUSION: Individual bulls showed variation in traits measured over age. The result of the study could be utilized in suggesting suitable management plans to achieve the desired profit by improving semen quality in Gir bulls.

The State of School Infrastructure in the Assembly Constituencies of Rural India: Analysis of 11 Census Indicators from Pre-Primary to Higher Education.

In India, assembly constituencies (ACs), represented by elected officials, are the primary geopolitical units for state-level policy development. However, data on social indicators are traditionally reported and analyzed at the district level, and are rarely available for ACs. Here, we combine village-level data from the 2011 Indian Census and AC shapefiles to systematically derive AC-level estimates for the first time. We apply this methodology to describe the distribution of 11 education infrastructures-ranging from pre-primary school to senior secondary school-across rural villages in 3773 ACs. We found high variability in access to higher education infrastructures and low variability in access to lower education variables. For 40.3% (25th percentile) to 79.7% (75th percentile) of villages in an AC, the nearest government senior secondary school was >5 km away, whereas the nearest government primary school was >5 km away in just 0% (25th percentile) to 1.9% (75th percentile) of villages in an AC. The states of Manipur, Arunachal Pradesh, and Bihar showed the greatest within-state variation in access to education infrastructures. We present a novel analysis of access to education infrastructure to inform AC-level policy, and demonstrate how geospatial and Census data can be leveraged to derive AC-level estimates for any population health and development indicators collected in the Census at the village level.

Efficient inhibition of foot-and-mouth disease virus replication in vitro by artificial microRNA targeting 3D polymerase.

Foot-and-mouth disease (FMD) is a devastating acute viral disease of livestock with cloven hooves. Among various therapeutic control measures, RNA interference (RNAi) is one of the methods being explored to inhibit FMD virus replication and spread. The RNAi is achieved by short hairpin RNAs or artificial microRNAs (amiRNAs). Utility of amiRNAs as antiviral, targeting conserved regions of the viral genome is gaining importance. However, delivery of miRNA in vivo is still a challenge. In this study, the efficacy of amiRNAs in preventing FMD virus replication in a permissive cell culture system was investigated, by generating stable cell lines expressing amiRNAs targeting three functional regions of the FMD virus (FMDV) genome (IRES, 3B3 and 3D). The results showed that amiRNA targeting 3D polymerase is relatively more efficient. However, expression of multiple microRNAs targeting the three regions did not exhibit additive effect. The data suggest that 3D specific miRNA is a potential valid strategy in developing novel antiviral measures against FMDV infection. Keywords: artificial microRNA; foot-and-mouth disease virus; virus inhibition.

Micro-geographic targeting for precision public policy: Analysis of child sex ratio across 587,043 census villages in India, 2011.

Child sex ratio (CSR) is a marker of disproportionate sex ratio at birth and discriminatory practices that lead to differential survival in early childhood by sex. We used the 2011 Census on rural India to present the first local analysis of CSR across 587,043 villages. In our multilevel analysis considering villages, tehsils, districts, and states/union territories, we found 96% of the total variation in CSR to be attributed to villages. About 39% of the villages were 'boy' areas (CSR≤88 girls per 100 boys) and another 12% had deficits in girls (88 < CSR≤93), while 11% fell in the normal range of CSR (93103). The magnitude of local variation in CSR was heterogeneous across states/union territories and districts. Our findings provide timely evidence to inform localized programmes like Beti Bachao, Beti Padhao to be implemented with greater precision.

Estimating the burden of child malnutrition across parliamentary constituencies in India: A methodological comparison.

In India, data on key developmental indicators used to formulate policies and interventions are routinely available for the administrative unit of districts but not for the political unit of parliamentary constituencies (PC). Recently, Swaminathan et al. proposed two methodologies to generate PC estimates using randomly displaced GPS locations of the sampling clusters ('direct') and by building a crosswalk between districts and PCs using boundary shapefiles ('indirect'). We advance these methodologies by using precision-weighted estimations based on hierarchical logistic regression modeling to account for the complex survey design and sampling variability. We exemplify this application using the latest National Family Health Survey (NFHS, 2016) to generate PC-level estimates for two important indicators of child malnutrition - stunting and low birth weight - that are being monitored by the Government of India for the National Nutrition Mission targets. Overall, we found a substantial variation in child malnutrition across 543 PCs. The different methodologies yielded highly consistent estimates with correlation ranging r = 0.92-0.99 for stunting and r = 0.81-0.98 for low birth weight. For analyses involving data with comparable nature to the NFHS (i.e., complex data structure and possibility to identify a potential PC membership), modeling for precision-weighted estimates and direct methodology are preferable. Further field work and data collection at the PC level are necessary to accurately validate our estimates. An ideal solution to overcome this gap in data for PCs would be to make PC identifiers available in routinely collected surveys and the Census.

Developing institutions for cancer care in low-income and middle-income countries: from cancer units to comprehensive cancer centres.

Global cancer centres operate across different sizes, scales, and ecosystems. Understanding the essential aspects of the creation, organisation, accreditation, and activities within these settings is crucial for developing an affordable, equitable, and quality cancer care, research, and education system. Robust guidelines are scarce for cancer units, cancer centres, and comprehensive cancer centres in low-income and middle-income countries. However, some robust examples of the delivery of complex cancer care in centres in emerging economies are available. Although it is impossible to create an optimal system to fit the unique needs of all countries for the delivery of cancer care, we summarise what has been published about the development and management of cancer centres in low-income and middle-income countries so far and highlight the need for clinical and political leadership.

Host serum microRNA profiling during the early stage of foot-and-mouth disease virus infection.

Foot-and-mouth disease virus (FMDV) causes a highly contagious infection in cloven-hoofed animals, with many outbreaks in the developing world. MicroRNAs (miRNAs) are non-coding RNAs that regulate antiviral defence by post-transcriptional regulation of gene expression. In this study, the host miRNA response following FMDV infection was investigated in cattle, a natural host for FMDV. A significant alteration in serum miRNA expression was detected at early stages of infection. Compared to prior to infection, on day 2 postinfection (PI), 119 miRNAs were upregulated, of which 39 were significantly upregulated (P < 0.05). Gene target prediction and pathway enrichment analysis suggested that upregulated miRNAs target innate immune signalling pathways, suggesting a homeostasis effect, possibly to limit inappropriate immune responses. Further, for the significantly upregulated miRNAs, nine miRNA recognition elements were identified in the genome sequence of FMDV serotype O, which was used for infection. The antiviral effect of four of these miRNAs was confirmed in a cell culture system. These data demonstrate that changes in miRNA expression occur during early pathogenesis, and the identification of possible miRNA targets genes could help in elucidating molecular events involved in virus-host interaction and thus could be useful in developing therapeutic strategies.

Genetic evaluation of growth in farmers' flocks of Madras Red sheep under long-term selection in a group breeding scheme.

The Network Project on Sheep Improvement (NWPSI)-Madras Red field unit is a group breeding scheme involving 198 farmers' flocks of Madras Red sheep in which selection for growth traits and rotation of rams have been practised for over two decades. Growth data collected from these flocks were used to evaluate the performance and understand the direct and expected responses to selection based on genetic parameters. The body weight at birth (BW), weaning weight (WW), 6-month weight (6W), 9-month weight (9W), 12-month weight (YW), pre-weaning average daily gain (ADG1, birth to 3 months), post-weaning ADG2 (3-6 months), ADG3 (6-9 months), ADG4 (9-12 months) and ADG5 (3-12 months) were 2.67, 10.05, 14.56, 18.36, 21.36, 80.13, 49.05, 43.00, 34.21 and 41.18 g, respectively. Univariate analyses were carried out using animal and sire models to estimate variance components. Heritability obtained from animal model for BW was 0.36 and the values for other body weight traits were almost unity. Heritability estimate for pre-weaning ADG1 was 0.31. Very high genetic variability was observed in spite of long-term selection and this sustenance of variability is one of the main advantages of a group breeding scheme, combining several flocks of smaller size. An increasing genetic and phenotypic trend was noticed for almost all the traits studied. The expected responses calculated based on genetic parameters also indicated scope for improvement.

Recombinant human adenovirus-5 expressing capsid proteins of Indian vaccine strains of foot-and-mouth disease virus elicits effective antibody response in cattle.

Recombinant adenovirus-5 vectored foot-and-mouth disease constructs (Ad5- FMD) were made for three Indian vaccine virus serotypes O, A and Asia 1. Constructs co-expressing foot-and- mouth disease virus (FMDV) capsid and viral 3C protease sequences, were evaluated for their ability to induce a neutralizing antibody response in indigenous cattle (Bos indicus). Purified Ad5-FMD viruses were inoculated in cattle as monovalent (5×109 pfu/animal) or trivalent (5×109 pfu/animal per serotype) vaccines. Animals vaccinated with monovalent Ad5-FMD vaccines were boosted 63days later with the same dose. After primary immunization, virus neutralization tests (VNT) showed seroconversion in 83, 67 and 33% of animals vaccinated with Ad5-FMD O, A and Asia 1, respectively. Booster immunization elicited seroconversion in all of the animals (100%) in the monovalent groups. When used in a trivalent form, the Ad5-FMD vaccine induced neutralizing antibodies in only 33, 50 and 16% of animals against serotypes O, A and Asia 1, respectively on primo-vaccination, and titers were significantly lower than when the same vectors were used in monovalent form. Neutralizing antibody titers differed by serotype for both Ad5-FMD monovalent and trivalent vaccines, with Asia 1 serotype inducing the lowest titers. Antibody response to Ad5 vector in immunized cattle was also assessed by VNT. It appeared that the vector immunity did not impact the recall responses to expressed FMDV antigens on booster immunization. In summary, the study suggested that the recombinant Ad5-FMD vaccine has a potential use in monovalent form, while its application in multivalent form is not currently encouraging.

Association of heat stress protein 90 and 70 gene polymorphism with adaptability traits in Indian sheep (Ovis aries).

Heat stress proteins assist cellular proteins in the acquisition of native structure. The present research was conducted to study how thermo-tolerance is modulated by HSP90 and HSP70 gene polymorphism and its association with hemato-physio-biochemical parameters, supported by their expression profiles in Chokla, Magra, Marwari, and Madras Red sheep breeds. Least square analysis revealed significant effect (P < 0.05) of season and breed on all the physiological parameters, i.e., temperature, respiratory rate, and pulse rate (a.m. and p.m.), as well as hematological parameters like Hb, packed cell volume, total erythrocyte count (TEC), neutrophil/lymphocyte (N/L) ratio, and total leukocyte count (TLC). There was a significant influence (P < 0.05) of breed on biochemical parameters such as glucose, SGOT, phosphorous, triglyceride, and cholesterol. Eight fragments were amplified and sequenced in HSP90, and 70 genes and 13 single-nucleotide polymorphisms (SNPs) were identified. Tetra-primer amplification refractory mutation system PCR, PCR-RFLP, and allele-specific PCR genotyping protocols were developed for large-scale genotyping of five SNPs. A significant difference (P < 0.05) of rectal temperature (a.m.), respiratory rate (p.m.), triglyceride, and total protein was observed at SNP01; albumin at SNP2; pulse rate (p.m.) at SNP3; and rectal temperature (p.m.), pulse rate (p.m.), Hb (g/dL), and N/L ratio at SNP4 and TLC at SNP5. Gene expression analysis revealed higher expression in less adapted animals with Madras Red < Magra < Chokla < Marwari expression pattern [corrected]. Predominant allele was found to be superior in most of the SNPs (SNP1-4) indicating the selection acting in directional manner (positive selection). Finally, it is concluded that TACCA haplotype combination of SNP1-SNP2-SNP3-SNP4-SNP5 might be of some selection advantage for the identification of animals more adaptable to heat stress.

Proteasome Inhibitor Carfilzomib-Based Therapy for Antibody-Mediated Rejection of the Pulmonary Allograft: Use and Short-Term Findings.

We present this observational study of lung transplant recipients (LTR) treated with carfilzomib (CFZ)-based therapy for antibody-mediated rejection (AMR) of the lung. Patients were considered responders to CFZ if complement-1q (C1q)-fixing ability of their immunodominant (ID) donor-specific anti-human leukocyte antibody (DSA) was suppressed after treatment. Treatment consisted of CFZ plus plasma exchange and immunoglobulins. Fourteen LTRs underwent CFZ for 20 ID DSA AMR. Ten (71.4%) of LTRs responded to CFZ. DSA IgG mean fluorescence intensity (MFI) fell from 7664 (IQR 3230-11 874) to 1878 (653-7791) after therapy (p = 0.001) and to 1400 (850-8287) 2 weeks later (p = 0.001). DSA C1q MFI fell from 3596 (IQR 714-14 405) to <30 after therapy (p = 0.01) and <30 2 weeks later (p = 0.02). Forced expiratory volume in 1s ( FEV1 ) fell from mean 2.11 L pre-AMR to 1.92 L at AMR (p = 0.04). FEV1 was unchanged after CFZ (1.91 L) and subsequently rose to a maximum of 2.13 L (p = 0.01). Mean forced expiratory flow during mid forced vital capacity (25-75) (FEF25-75 ) fell from mean 2.5 L pre-AMR to 1.95 L at AMR (p = 0.01). FEF25-75 rose after CFZ to 2.54 L and reached a maximum of 2.91 L (p = 0.01). Responders had less chronic lung allograft dysfunction or progression versus nonresponders (25% vs. 83%, p = 0.04). No deaths occurred within 120 days and 7 patients died post CFZ therapy of allograft failure. Larger prospective interventional studies are needed to further describe the benefit of CFZ-based therapy for pulmonary AMR.

A liver microphysiological system of tumor cell dormancy and inflammatory responsiveness is affected by scaffold properties.

Distant metastasis is the major cause of breast cancer-related mortality, commonly emerging clinically after 5 or more years of seeming 'cure' of the primary tumor, indicating a quiescent dormancy. The lack of relevant accessible model systems for metastasis that recreate this latent stage has hindered our understanding of the molecular basis and the development of therapies against these lethal outgrowths. We previously reported on the development of an all-human 3D ex vivo hepatic microphysiological system that reproduces several features of liver physiology and enables spontaneous dormancy in a subpopulation of breast cancer cells. However, we observed that the dormant cells were localized primarily within the 3D tissue, while the proliferative cells were in contact with the polystyrene scaffold. As matrix stiffness is known to drive inflammatory and malignant behaviors, we explored the occurrence of spontaneous tumor dormancy and inflammatory phenotype. The microphysiological system was retrofitted with PEGDa-SynKRGD hydrogel scaffolding, which is softer and differs in the interface with the tissue. The microphysiological system incorporated donor-matched primary human hepatocytes and non-parenchymal cells (NPCs), with MDA-MB-231 breast cancer cells. Hepatic tissue in hydrogel scaffolds secreted lower levels of pro-inflammatory analytes, and was more responsive to inflammatory stimuli. The proportion of tumor cells entering dormancy was markedly increased in the hydrogel-supported tissue compared to polystyrene. Interestingly, an unexpected differential response of dormant cells to varying chemotherapeutic doses was identified, which if reflective of patient pathophysiology, has important implications for patient dosing regimens. These findings highlight the metastatic microphysiological system fitted with hydrogel scaffolds as a critical tool in the assessment and development of therapeutic strategies to target dormant metastatic breast cancer.