Non-Classical Swine Leukocyte Antigens SLA-6, -7, and -8, Are Xenoantigens for Some Waitlisted Patients.

Attack of donor tissues by pre-formed anti-pig antibodies is well known to cause graft failure in xenotransplantation. Genetic engineering of porcine donors to eliminate targets of these pre-formed antibodies coupled with advances in immunosuppressive medicines have now made it possible to achieve extended survival in the pre-clinical pig-to-non-human primate model. Despite these improvements, antibodies remain a risk over the lifetime of the transplant, and many patients continue to have pre-formed donor-specific antibodies even to highly engineered pigs. While therapeutics exist that can help mitigate the detrimental effects of antibodies, they act broadly potentially dampening beneficial immunity. Identifying additional xenoantigens may enable more targeted approaches, such as gene editing, to overcome these challenges by further eliminating antibody targets on donor tissue. Because we have found that classical class I swine leukocyte antigens are targets of human antibodies, we now examine whether related pig proteins may also be targeted by human antibodies. We show here that non-classical class I swine leukocyte proteins (SLA-6, -7, -8) can be expressed at the surface of mammalian cells and act as antibody targets.

Late graft failure of pig-to-rhesus renal xenografts has features of glomerulopathy and recipients have anti-swine leukocyte antigen class I and class II antibodies.

Prolonged survival in preclinical renal xenotransplantation demonstrates that early antibody mediated rejection (AMR) can be overcome. It is now critical to evaluate and understand the pathobiology of late graft failure and devise new means to improve post xenograft outcomes. In renal allotransplantation the most common cause of late renal graft failure is transplant glomerulopathy-largely due to anti-donor MHC antibodies, particularly anti-HLA DQ antibodies. We evaluated the pig renal xenograft pathology of four long-surviving (>300 days) rhesus monkeys. We also evaluated the terminal serum for the presence of anti-SLA class I and specifically anti-SLA DQ antibodies. All four recipients had transplant glomerulopathy and expressed anti-SLA DQ antibodies. In one recipient tested for anti-SLA I antibodies, the recipient had antibodies specifically reacting with two of three SLA I alleles tested. These results suggest that similar to allotransplantation, anti-MHC antibodies, particularly anti-SLA DQ, may be a barrier to improved long-term xenograft outcomes.

Intimate partner violence and suicide mortality: a cross-sectional study using machine learning and natural language processing of suicide data from 43 states.

INTRODUCTION: Intimate partner violence (IPV) is associated with suicidal ideation, yet it remains unclear how often IPV precipitates suicide mortality. To overcome limitations with national data, we applied novel methods to: (1) document the prevalence of IPV-related suicide in the USA and (2) identify correlates for IPV-related suicide. METHODS: Using National Violent Death Reporting System data (NVDRS, 2015-2019, n=1 30 550), we recorded IPV circumstances (yes/no) by leveraging prior textual reviews of death narratives and applying a validated natural language processing tool. We could not systematically differentiate IPV perpetration versus victimisation given limited details in NVDRS. Logistic regression compared IPV-related suicides with referent group suicides (no evidence of IPV), stratified by sex. RESULTS: 7.1% of suicides were IPV related (n=9210), most were isolated suicide events (82.8%, n=7625; ie, not homicide suicide). There were higher odds of IPV circumstances when the decedent had civil legal problems (aOR for men: 3.6 (3.3 to 3.9), aOR for women: 2.6 (2.2 to 3.2)), criminal legal problems (aOR men: 2.3 (2.2 to 2.5), aOR for women: 1.7 (1.4 to 2.1)), or used a firearm (aOR men: 1.9 (1.8 to 2.0), aOR for women: 1.9 (1.7 to 2.1)). There were lower odds of IPV circumstances when the decedent had a current mental health problem (aOR men: 0.7 (0.7 to 0.8), aOR for women: 0.7 (0.6 to 0.8)). CONCLUSIONS: IPV circumstances contribute to a notable proportion of suicides. IPV-related suicides are distinct from other suicide deaths. Targeted suicide screening and intervention in IPV settings may be beneficial for prevention.

Xenoreactive antibodies in α-granules of human platelets bind pig liver endothelial cells.

Pig liver xenotransplantation is limited by a thrombocytopenic coagulopathy that occurs immediately following graft reperfusion. In vitro and ex vivo studies from our lab suggested that the thrombocytopenia may be the result of a species incompatibility in platelet glycosylation. Realization that platelet α-granules contain antibodies caused us to reevaluate whether the thrombocytopenia in liver xenotransplantation could occur because IgM and IgG from inside platelet α-granules bound to pig liver sinusoidal endothelial cells (LSECs). Our in vitro analysis of IgM and IgG from inside α-granules showed that platelets do carry xenoreactive antibodies that can bind to known xenoantigens. This study suggests that thrombocytopenia occurring following liver xenotransplantation could occur because of xenoreactive antibodies tethering human platelets to the pig LSEC enabling the platelet to be phagocytosed. These results suggest genetic engineering strategies aimed at reducing xenoantigens on the surface of pig LSEC will be effective in eliminating the thrombocytopenia that limits survival in liver xenotransplantation.

Detecting intimate partner violence circumstance for suicide: development and validation of a tool using natural language processing and supervised machine learning in the National Violent Death Reporting System.

BACKGROUND: Intimate partner violence (IPV) victims and perpetrators often report suicidal ideation, yet there is no comprehensive national dataset that allows for an assessment of the connection between IPV and suicide. The National Violent Death Reporting System (NVDRS) captures IPV circumstances for homicide-suicides (<2% of suicides), but not single suicides (suicide unconnected to other violent deaths; >98% of suicides). OBJECTIVE: To facilitate a more comprehensive understanding of the co-occurrence of IPV and suicide, we developed and validated a tool that detects mentions of IPV circumstances (yes/no) for single suicides in NVDRS death narratives. METHODS: We used 10 000 hand-labelled single suicide cases from NVDRS (2010-2018) to train (n=8500) and validate (n=1500) a classification model using supervised machine learning. We used natural language processing to extract relevant information from the death narratives within a concept normalisation framework. We tested numerous models and present performance metrics for the best approach. RESULTS: Our final model had robust sensitivity (0.70), specificity (0.98), precision (0.72) and kappa values (0.69). False positives mostly described other family violence. False negatives used vague and heterogeneous language to describe IPV, and often included abusive suicide threats. IMPLICATIONS: It is possible to detect IPV circumstances among singles suicides in NVDRS, although vague language in death narratives limited our tool's sensitivity. More attention to the role of IPV in suicide is merited both during the initial death investigation processes and subsequent NVDRS reporting. This tool can support future research to inform targeted prevention.

Intimate partner violence victimization during pregnancy increases risk of postpartum depression among urban adolescent mothers in South Africa.

BACKGROUND: It is estimated that 38.8% of mothers develop postpartum depression (PPD) in South Africa. While empirical evidence documents an association between intimate partner violence (IPV) victimization in pregnancy and PPD among adult women, the association has been underexamined among adolescent mothers (< 19 years). The study's purpose is to examine whether IPV victimization during pregnancy is associated with PPD among adolescent mothers. METHODS: Adolescent mothers (14-19 years) were recruited at a regional hospital's maternity ward in KwaZulu Natal, South Africa between July 2017-April 2018. Participants completed behavioral assessments at two visits (n = 90): baseline (up to 4 weeks postpartum) and follow-up (6-9 weeks postpartum, when PPD is typically assessed). The WHO modified conflict tactics scale was used to create a binary measure of any physical and/or psychological IPV victimization that occurred during pregnancy. Participants with scores ≥ 13 on the Edinburgh Postpartum Depression Scale (EPDS) were classified as having symptoms of PPD. We used a modified Poisson regression with robust standard errors to assess PPD in association with IPV victimization during pregnancy, controlling for relevant covariates. RESULTS: Nearly one-half (47%) of adolescent mothers reported symptoms of PPD by 6-9 weeks post-delivery. Further, IPV victimization during pregnancy was highly prevalent (40%). Adolescent mothers who reported IPV victimization during pregnancy had marginally higher risk of PPD at follow-up (RR: 1.50, 95 CI: 0.97-2.31; p = 0.07). The association was strengthened and significant in covariate-adjusted analysis (RR: 1.62, 95 CI: 1.06-2.49; p = 0.03). CONCLUSIONS: Poor mental health was common among adolescent mothers, and IPV victimization during pregnancy was associated with PPD risk among adolescent mothers. Implementing IPV and PPD routine screenings during the perinatal period may aid in identifying adolescent mothers for IPV and PPD interventions and treatment. With the high prevalence of IPV and PPD in this vulnerable population and the potential negative impact on maternal and infant outcomes, interventions to reduce IPV and PPD are needed to improve adolescent mothers' well-being and their baby's health.

BACKGROUND: More than one-third of adult mothers experience postpartum depression (PPD) in South Africa and intimate partner violence (IPV) victimization is a strong risk factor of PPD for adult mothers. However, there are no studies on adolescent mothers that look at the link between IPV victimization and PPD. This paper aims to examine whether IPV victimization during pregnancy is associated with PPD among adolescent South African mothers. METHODS: We had 90 adolescent mothers (aged 14­19 years old) complete an initial survey between delivery and 4 weeks postpartum to collect information on IPV during their pregnancy. Participants completed an additional survey between 6 and 9 week postpartum to collect information on the symptoms of PPD. RESULTS: Nearly one-half (47%) of adolescent mothers reported symptoms of PPD by 6­9 weeks post-delivery. Report of IPV victimization during pregnancy was also very high (40%). Adolescent mothers who experienced IPV victimization during pregnancy were more likely to report symptoms of PPD between 6 and 9 weeks postpartum. CONCLUSIONS: PPD and IPV was very common in our sample, and IPV victimization during pregnancy was linked to PPD among adolescent mothers. Having routine screenings during pregnancy and postpartum period can identify adolescent mothers IPV and PPD interventions and treatment. With the high reports of IPV and PPD in this sample and the potential negative impact on maternal and infant outcomes, interventions to reduce IPV and PPD are needed to improve adolescent mothers' well-being and their baby's health.

'She showed me a new path, a way forward': exploring how navigation influences mental health among Guatemalan gay and bisexual men living with HIV.

Mental health problems, including anxiety and depression, are a common comorbidity among gay, bisexual and other men who have sex with men (GBMSM) living with HIV. Informed by social support theory, health navigation is a strengths-based intervention that has been demonstrated to improve HIV care outcomes. The purpose of this study was to explore how health navigation influences the mental health of GBMSM living with HIV. We analyzed longitudinal qualitative in-depth interviews conducted with GBMSM (n = 29) in a 12-month multi-component intervention to improve HIV care outcomes, including health navigation. We used narrative and thematic analytic approaches to identify salient themes, including if and how themes changed over time. Participants described that navigator support helped them maintain good mental health, prevent crises and respond to crises. Navigator support included providing motivational messaging, facilitating participants' control over their health and improving access to care, which aided with supporting mental health. Navigators also responded to acute crises by providing guidance for those newly diagnosed with HIV and support for those experiencing critical life events. Participants emphasized the importance of feeling heard and valued by their navigators and gaining hope for the future as key to their wellbeing. In conclusion, health navigation may be an effective intervention for promoting mental health among GBMSM living with HIV. Additional research is needed to examine mediating pathways between navigation and mental health, including informational support, or if navigator support moderates the relationship between stressors and mental health outcomes for GBMSM.

A noncanonical GATA transcription factor of Entamoeba histolytica modulates genes involved in phagocytosis.

In this paper, we explored the presence of GATA in Entamoeba histolytica and their function as regulators of phagocytosis-related genes. Bioinformatics analyses evidenced a single 579 bp sequence encoding for a protein (EhGATA), smaller than GATA factors of other organisms. EhGATA appeared phylogenetically close to Dictyostelium discoideum and Schistosoma mansoni GATA proteins. Its sequence predicts the presence of a zinc-finger DNA binding domain and an AT-Hook motif; it also has two nuclear localization signals. By transmission electron and confocal microscopy, anti-EhGATA antibodies revealed the protein in the cytoplasm and nucleus, and 65% of nuclear signal was in the heterochromatin. EhGATA recombinant protein and trophozoites nuclear extracts bound to GATA-DNA consensus sequence. By in silico scrutiny, 1,610 gene promoters containing GATA-binding sequences appeared, including Ehadh and Ehvps32 promoters, whose genes participate in phagocytosis. Chromatin immunoprecipitation assays showed that EhGATA interact with Ehadh and Ehvps32 promoters. In EhGATA-overexpressing trophozoites (NeoGATA), the Ehadh and Ehvps32 mRNAs amount was modified, strongly supporting that EhGATA could regulate their transcription. NeoGATA trophozoites exhibited rounded shapes, high proliferation rates, and diminished erythrophagocytosis. Our results provide new insights into the role of EhGATA as a noncanonical transcription factor, regulating genes associated with phagocytosis.

Anti-C5 Antibody Tesidolumab Reduces Early Antibody-mediated Rejection and Prolongs Survival in Renal Xenotransplantation.

OBJECTIVE: Pig-to-primate renal xenotransplantation is plagued by early antibody-mediated graft loss which precludes clinical application of renal xenotransplantation. We evaluated whether temporary complement inhibition with anti-C5 antibody Tesidolumab could minimize the impact of early antibody-mediated rejection in rhesus monkeys receiving pig kidneys receiving costimulatory blockade-based immunosuppression. METHODS: Double (Gal and Sda) and triple xenoantigen (Gal, Sda, and SLA I) pigs were created using CRISPR/Cas. Kidneys from DKO and TKO pigs were transplanted into rhesus monkeys that had the least reactive crossmatches. Recipients received anti-C5 antibody weekly for 70 days, and T cell depletion, anti-CD154, mycophenolic acid, and steroids as baseline immunosuppression (n = 7). Control recipients did not receive anti-C5 therapy (n = 10). RESULTS: Temporary anti-C5 therapy reduced early graft loss secondary to antibody-mediated rejection and improved graft survival (P < 0.01). Deleting class I MHC (SLA I) in donor pigs did not ameliorate early antibody-mediated rejection (table). Anti-C5 therapy did not allow for the use of tacrolimus instead of anti-CD154 (table), prolonging survival to a maximum of 62 days. CONCLUSION: Inhibition of the C5 complement subunit prolongs renal xenotransplant survival in a pig to non-human primate model.

IPV victimization in pregnancy increases postpartum STI incidence among adolescent mothers in Durban, South Africa.

Women, and specifically, adolescents, are at high risk of HIV and STIs during the postpartum period. Biological and behavioral factors contribute to adolescents' susceptibility. However, the influence of behavioral factors, like intimate partner violence (IPV), on postpartum STI acquisition has been understudied. The study's purpose is to determine whether IPV victimization during pregnancy predicts incident STIs in the first 6 months postpartum. Adolescent mothers (14-19 years) were recruited at a township hospital's maternity ward near Durban. Adolescent mothers who were HIV-negative and had no laboratory-diagnosed STIs at baseline (6 weeks postpartum) were included in the analysis (n = 61). We used a modified Poisson regression with robust standard errors to assess differences in postpartum STI risk by IPV victimization during pregnancy controlling for covariates. At baseline, 25 (41%) adolescent mothers reported IPV victimization during pregnancy. Adolescent mothers who reported IPV during pregnancy were at higher risk of receiving an STI diagnoses at 6 months postpartum (aRR: 4.43; 95% CI: 1.31-14.97). Our findings heighten understanding of HIV risk among a vulnerable subset of adolescent girls: adolescent mothers. Non-combined interventions that help young mothers and their partners navigate partnership dynamics to reduce IPV and STIs are needed to reduce HIV risk.

Examining the Biosynthesis and Xenoantigenicity of Class II Swine Leukocyte Antigen Proteins.

Genetically engineered pig organs could provide transplants to all patients with end-stage organ failure, but Ab-mediated rejection remains an issue. This study examines the class II swine leukocyte Ag (SLA) as a target of epitope-restricted Ab binding. Transfection of individual α- and ß-chains into human embryonic kidney cells resulted in both traditional and hybrid class II SLA molecules. Sera from individuals on the solid organ transplant waiting list were tested for Ab binding and cytotoxicity to this panel of class II SLA single-Ag cells. A series of elution studies from an SLA-DQ cell line were performed. Our results indicate that human sera contain Abs specific for and cytotoxic against class II SLA. Our elution studies revealed that sera bind the SLA-DQ molecule in an epitope-restricted pattern. Site-specific mutation of one of these epitopes resulted in statistically decreased Ab binding. Humans possess preformed, specific, and cytotoxic Abs to class II SLA that bind in an epitope-restricted fashion. Site-specific epitope mutagenesis may decrease the Ab binding of highly sensitized individuals to pig cells.

Endoscopic stenting for gastroduodenal outlet obstruction of a malignant origin, real life experience in a single center.

AIM: to evaluate the safety and effectiveness of self-expandable metal stent placement for malignant gastric outlet obstruction (GOO). METHODS: a retrospective, analytic cohort study at a single, tertiary-care center. RESULTS: thirty-six patients that underwent stent placement for GOO of malignant origin were identified during the study period. Technical success was achieved in 36 (100 %) patients and clinical success was achieved in 31 patients (86.1 %). Before the procedure, 17 (54.8 %) patients had a gastric outlet obstruction score (GOOSS) of 0, which is a complete inability of oral intake. Twenty-three patients were alive 30 days after the procedure, two (8.6 %) patients had a GOOSS of 1, ten (43.3 %) had a GOOSS of 2 and eleven (47.9 %) had a GOOSS of 3. Abdominal pain was present in all 31 patients before the procedure and only seven (22.6 %) patients continued with abdominal pain 24 hours after the procedure. During follow-up, ten (30.3 %) patients developed complications related to the stents and none of them was fatal. Additional therapy due to partial occlusion of the stent was necessary in three patients. The stents functional duration had a median of 72 days (IQR 25-75 15-105 days) and was closely related to overall survival. CONCLUSION: palliative stenting for gastroduodenal obstruction is a safe, feasible and effective therapy to treat patients with malignant gastric outlet obstruction.

Examining epitope mutagenesis as a strategy to reduce and eliminate human antibody binding to class II swine leukocyte antigens.

Xenotransplantation of pig organs into people may help alleviate the critical shortage of donors which faces organ transplantation. Unfortunately, human antibodies vigorously attack pig tissues preventing the clinical application of xenotransplantation. The swine leukocyte antigens (SLA), homologs of human HLA molecules, can be xenoantigens. SLA molecules, encoded by genes in the pig major histocompatibility complex, contribute to protective immune responses in pig. Therefore, simply inactivating them through genome engineering could reduce the ability of the human immune system to surveil transplanted pig organs for infectious disease or the development of neoplasms. A potential solution to this problem is to identify and modify epitopes in SLA proteins to eliminate their contribution to humoral xenoantigenicity while retaining their biosynthetic competence and ability to contribute to protective immunity. We previously showed that class II SLA proteins were recognized as xenoantigens and mutating arginine at position 55 to proline, in an SLA-DQ beta chain, could reduce human antibody binding. Here, we extend these observations by creating several additional point mutants at position 55. Using a panel of monoclonal antibodies specific for class II SLA proteins, we show that these mutants remain biosynthetically competent. Examining antibody binding to these variants shows that point mutagenesis can reduce, eliminate, or increase antibody binding to class II SLA proteins. Individual mutations can have opposite effects on antibody binding when comparing samples from different people. We also performed a preliminary analysis of creating point mutants near to position 55 to demonstrate that manipulating additional residues also affects antibody reactivity.

Treatment of gastrointestinal bleeding with hemostatic powder (TC-325): a multicenter study.

INTRODUCTION: Hemostatic powder (TC-325) is a new tool for treatment of gastrointestinal bleeding that allows the treatment of large surfaces with active bleeding. The aim was to describe the initial success of TC-325 for the control of GI bleeding. MATERIALS AND METHODS: We did a multicenter cohort study with patients admitted to the endoscopy service for GI bleeding. A format was generated to standardize the information obtained in each center. It was determined whether this treatment had been used as a single therapy or as a combination therapy. Descriptive statistics with medians and ranges, or averages with SD according to distribution. RESULTS: Eighty-one patients with 104 endoscopic procedures were included. The median number of endoscopic procedures was 1 (1-3). In the first procedure, the initial success rate was 98.8% (n = 80), failure rate was 1.2% (n = 1), and rebleeding rate was 20% (n = 16). The majority of rebleeding cases occurred within the first 3 days (12/16, 75%). There was no association between rebleeding and etiology (malignant or benign; P = 0.6). In first procedure, 44 (54%) cases had monotherapy with TC-325 and 37 (46%) cases had a combined endoscopic therapy. There were no differences in initial success or rebleeding rates when TC-325 was used as monotherapy versus combined therapy (P = 0.7). The mortality rate was 4% (3/81). CONCLUSION: TC-325 is effective for achieving initial control of bleeding in patients with different GI etiologies. The rate of bleeding recurrence is considerable in both patients with benign and malignant etiology.

Xenoantigen Deletion and Chemical Immunosuppression Can Prolong Renal Xenograft Survival.

OBJECTIVE: Xenotransplantation using pig organs could end the donor organ shortage for transplantation, but humans have xenoreactive antibodies that cause early graft rejection. Genome editing can eliminate xenoantigens in donor pigs to minimize the impact of these xenoantibodies. Here we determine whether an improved cross-match and chemical immunosuppression could result in prolonged kidney xenograft survival in a pig-to-rhesus preclinical model. METHODS: Double xenoantigen (Gal and Sda) knockout (DKO) pigs were created using CRISPR/Cas. Serum from rhesus monkeys (n = 43) was cross-matched with cells from the DKO pigs. Kidneys from the DKO pigs were transplanted into rhesus monkeys (n = 6) that had the least reactive cross-matches. The rhesus recipients were immunosuppressed with anti-CD4 and anti-CD8 T-cell depletion, anti-CD154, mycophenolic acid, and steroids. RESULTS: Rhesus antibody binding to DKO cells is reduced, but all still have positive CDC and flow cross-match. Three grafts were rejected early at 5, 6, and 6 days. Longer survival was achieved in recipients with survival to 35, 100, and 435 days. Each of the 3 early graft losses was secondary to IgM antibody-mediated rejection. The 435-day graft loss occurred secondary to IgG antibody-mediated rejection. CONCLUSIONS: Reducing xenoantigens in donor pigs and chemical immunosuppression can be used to achieve prolonged renal xenograft survival in a preclinical model, suggesting that if a negative cross-match can be obtained for humans then prolonged survival could be achieved.

CRISPR/Cas and recombinase-based human-to-pig orthotopic gene exchange for xenotransplantation.

BACKGROUND: Tools for genome editing in pigs are improving rapidly so that making precise cuts in DNA for the purposes of deleting genes is straightforward. Development of means to replace pig genes with human genes with precision is very desirable for the future development of donor pigs for xenotransplantation. MATERIALS AND METHODS: We used Cas9 to cut pig thrombomodulin (pTHBD) and replace it with a plasmid containing a promoterless antibiotic selection marker and the exon for human thrombomodulin. PhiC31 recombinase was used to remove the antibiotic selection marker to create porcine aortic endothelial cells expressing human instead of pTHBD, driven by the endogenous pig promoter. RESULTS: The promoterless selection cassette permitted efficient enrichment of cells containing correctly inserted transgene. Recombinase treatment of selected cells excised the resistance marker permitting expression of the human transgene by the endogenous pTHBD promoter. Gene regulation was maintained after gene replacement because pig endogenous promoter was kept intact in the correct position. CONCLUSIONS: Cas9 and recombinase technology make orthotopic human for pig gene exchange feasible and pave the way for creation of pigs with human genes that can be expressed in the appropriate tissues preserving gene regulation.

Silencing porcine genes significantly reduces human-anti-pig cytotoxicity profiles: an alternative to direct complement regulation.

UNLABELLED: The future of solid organ transplantation is challenged by an increasing shortage of available allografts. Xenotransplantation of genetically modified porcine organs offers an answer to this problem. Strategies of genetic modification have 'humanized' the porcine model towards clinical relevance. Most notably, these approaches have aimed at either antigen reduction or human transgene expression. The object of this study was to evaluate the relative effects of both antigen reduction and direct complement regulation on the human-anti-porcine complement dependent cytotoxicity response. Genetically modified animals were created through CRISPR/Cas9-directed mutation and human transgene delivery. Pigs doubly deficient in GGTA1 and CMAH genes were compared to pigs of the same background that expressed a human complement regulatory protein (hCRP). A third animal was made deficient in GGTA1, CMAH and B4GalNT2 gene expression. Cells from these animals were subjected to measures of human antibody binding and antibody-mediated complement-dependent cytotoxicity by flow cytometry. Human IgG and IgM antibody binding was unchanged between the double knockout and the transgenic hCRP double knockout pig. IgG and IgM binding was reduced by 49.1 and 43.2 % respectively by silencing the B4GalNT2 gene. Compared to the double knockout, human anti-porcine cytotoxicity was reduced by 8 % with the addition of a hCRP (p = .032); It was reduced by 21 % with silencing the B4GalNT2 gene (p = .012). CONCLUSIONS: Silencing the GGTA1, CMAH and B4GalNT2 genes in pigs achieved a significant antigen reduction. Changing the porcine carbohydrate profile effectively mediates human antibody-mediated complement dependent cytoxicity.

Creating class I MHC-null pigs using guide RNA and the Cas9 endonuclease.

Pigs are emerging as important large animal models for biomedical research, and they may represent a source of organs for xenotransplantation. The MHC is pivotal to the function of the immune system in health and disease, and it is particularly important in infection and transplant rejection. Pigs deficient in class I MHC could serve as important reagents to study viral immunity as well as allograft and xenograft rejection. In this study, we report the creation and characterization of class I MHC knockout pigs using the Cas9 nuclease and guide RNAs. Pig fetal fibroblasts were genetically engineered using Cas9 and guide RNAs, and class I MHC(-) cells were then used as nuclear donors for somatic cell nuclear transfer. We produced three piglets devoid of all cell surface class I proteins. Although these animals have reduced levels of CD4(-)CD8(+) T cells in peripheral blood, the pigs appear healthy and are developing normally. These pigs are a promising reagent for immunological research.

Extended source model for diffusive coupling.

Motivated by the prevailing approach to diffusion coupling phenomena which considers point-like diffusing sources, we derived an analogous expression for the concentration rate of change of diffusively coupled extended containers. The proposed equation, together with expressions based on solutions to the diffusion equation, is intended to be applied to the numerical solution of systems exclusively composed of ordinary differential equations, however is able to account for effects due the finite size of the coupled sources.

The Effects of the Moms and Teens for Safe Dates Program on Dating Abuse: a Conditional Process Analysis.

Moms and Teens for Safe Dates (MTSD) is a dating abuse (DA) prevention program for teens exposed to domestic violence. In a randomized controlled trial (RCT), MTSD prevented certain types of DA victimization (psychological and physical) and perpetration (psychological and cyber) among teens with higher, but not lower, exposure to domestic violence. We built on these findings by using moderated mediation analysis to examine whether level of teen exposure to domestic violence conditioned the indirect effects of MTSD on these types of DA through targeted mediators. MTSD consisted of six mailed activity booklets. Mothers who had been former victims of domestic violence delivered the program to their teens. Mother and teen pairs were recruited into the RCT through community advertising and completed baseline and 6-month follow-up interviews (N = 277 pairs). As expected, MTSD had significant favorable effects for teens with higher but not lower exposure to domestic violence on several mediators that guided program content, including teen conflict management skills and mother-perceived severity of DA, self-efficacy for enacting DA prevention efforts, and comfort in communicating with her teen. MTSD had significant main effects on other mediators including teen feeling of family closeness and cohesion and mother-perceived susceptibility of her teen to DA. As expected, all significant indirect effects of MTSD on DA outcomes through mediators were for teens with higher exposure to domestic violence. Findings have implications for developing DA victimization and perpetration prevention programs for teens with high exposure to domestic violence.