NEG Groups Research Articles

S1582 Abnormal Glucose Breath Test Does Not Correlate With Patient’s Self-Reported Symptoms and Their Severity

Introduction: Glucose Hydrogen Breath Testing (GBT) is commonly used for diagnosing Small Intestinal Bacterial Overgrowth (SIBO). However, patients present with clinical symptoms (sx) and their predictive value for a positive (pos) GBT remains unclear. AIM: To assess the prevalence of self-reported GI sx profiles in adults with unexplained upper GI symptoms and correlate this with GBT. Methods: We retrospectively analyzed all adult patients presenting to a single medical center from 6/20 –12/21 for GBT. A pos GBT for SIBO was defined by the North American Consensus (≥20 ppm H2 increase over basal within 90 minutes (mins) and/or ≥10 ppm CH4 at any time). Pts completed a GI questionnaire assessing 17 key sxs: regurgitation (RG), chest pain (CP), heartburn (HB), belch (BH), abdominal pain (AP), bloating (BT), gas (GS), nausea (NA), vomit (VT), abdominal cramps (AC), indigestion (ID), distension (DN), fullness (FL), early satiety (ES), diarrhea (DR), constipation (CN), and fatigue (FT). Sxs measured by frequency (None, < 1/week (wk), 1/wk, ≥ 1/wk), intensity (None, Mild, Moderate, Severe), and duration (None, < 10 mins, 10 – 30 mins, ≥ 30 mins). Sub-groups, GERD, Dyspeptic, and IBS were constructed assessing the mean summation scores for Frequency, Intensity, and Duration between negative (neg) and pos GBTs. Univariate analysis and logistic regression analysis was performed. A P-value of < 0.05 was considered statistically significant. Results: We evaluated 184 pts, mean age of 51.7 (18 – 85), 78.3% female, 82.8% Caucasian, and mean BMI of 27.2. No significant differences were seen between GI sx frequency and intensity categories for pts with GBT neg and pos groups. A trend for a higher percentage of pos GBT pts reporting FL (P=0.09) and DR (P=0.07) in sx duration categories of 10 -30 mins (FL: 30.8% vs 27.5%; DR: 31.8% vs 23.4%) and ≥ 30 mins (FL: 51.3% vs 41.3%; DR: 37.2% vs 22.1%) was noted. Regression analysis showed a trend toward a pos GBT for VT frequency of Less than 1/wk: (P = 0.07; OR = 2.99; 95% CI: 0.91, 9.83) and NA intensity of Severe: (P = 0.09; OR = 2.24; 95% CI: 0.86, 5.84). No differences were seen between the GERD, Dyspeptic, and IBS sub-groups (Figure). Conclusion: The pre-test predictability of GI symptoms as it relates to a pos or neg GBT is poor. The yield for a pos GBT varies between 31%-55% and depends on age. In routine practice, when the index of clinical suspicion is high for SIBO, clinicians should consider objective testing with GBT rather than empiric antibiotic treatment.Figure 1.: (Left) Mean Summation Scores for Self-Reported Frequency, Intensity, and Duration Responses to GI Symptoms Relative to the Gastroesophageal Reflux Disease (GERD), Dyspeptic, and irritable bowel syndrome (IBS) Groups. (Right) Prevalence of Positive Glucose Breath Test (GBT) Among Age Cohorts and Glucose Breath Test Measurements

The differences of parasitemia and lungs size in malaria-associated acute respiratory distress syndrome (MA-ARDS) and non-MA-ARDS in mice infected with Plasmodium berghei ANKA

Malaria-associated acute respiratory distress syndrome (MA-ARDS) is characterized by extensive infiltration of leukocytes, microhemorrhages, vasogenic edema, changes in lung color, and a significant increase in the weight of the lung. This study was aimed to find out the differences in parasitemia and lung size in MA-ARDSand non-MA-ARDSin mice infected with Plasmodium berghei ANKA. Sixteen male BALB/c mice were infected with P. berghei ANKA, and daily parasitemia was observed on Giemsa-stained tail blood smears. Mice were sacrificed when parasitemia reached ±20%. Simultaneously eight uninfected mice were used as negative control (NEG). The statistical analysis was done using Kruskal Wallis, Mann Whitney U tests, and Spearman correlation test. The results showed that there were significant differences in parasitemia (p=0.001), weight (p=0.001), and lung length (p=0.021) between the MA-ARDS and non-MA-ARDS groups. Comparison of NEG and MA-ARDS resulted in a significant difference in lung size (p=0.05). When non-MA-ARDScompared with NEG groups, it showed a significant difference in lung width (p=0.001). However, there was no significant difference in lung weight and length (p&gt;0.05). Spearman correlation test showed that there was a strong correlation between parasitemia with weight (p=0.000), length (p=0.001), and lung width (p=0.017). The findings indicated that parasitemia played a role in the development of MA-ARDS in mice infected with P. berghei ANKA and influenced the size of the lung.

Measurable residual disease status and outcome of transplant in acute myeloid leukemia in second complete remission: a study by the acute leukemia working party of the EBMT

Measurable residual disease (MRD) prior to hematopoietic cell transplant (HCT) for acute myeloid leukemia (AML) in first complete morphological remission (CR1) is an independent predictor of outcome, but few studies address CR2. This analysis by the Acute Leukemia Working Party of the European Society for Blood and Marrow Transplantation registry assessed HCT outcomes by declared MRD status in a cohort of 1042 adult patients with AML CR2 at HCT. Patients were transplanted 2006–2016 from human leukocyte antigen (HLA) matched siblings (n = 719) or HLA 10/10 matched unrelated donors (n = 293). Conditioning was myeloablative (n = 610) or reduced-intensity (n = 432) and 566 patients (54%) had in-vivo T cell depletion. At HCT, 749 patients (72%) were MRD negative (MRD NEG) and 293 (28%) were MRD positive (MRD POS). Time from diagnosis to HCT was longer in MRD NEG than MRD POS patients (18 vs. 16 months (P < 0.001). Two-year relapse rates were 24% (95% CI, 21–28) and 40% (95% CI, 34–46) in MRD NEG and MRD POS groups (P < 0.001), respectively. Leukemia-free survival (LFS) was 57% (53–61) and 46% (40–52%), respectively (P = 0.001), but there was no difference in terms of overall survival. Prognostic factors for relapse and LFS were MRD NEG status, good risk cytogenetics, and longer time from diagnosis to HCT. In-vivo T cell depletion predicted relapse.

Heifers with positive genetic merit for fertility traits reach puberty earlier and have a greater pregnancy rate than heifers with negative genetic merit for fertility traits

This study investigated the hypothesis that dairy heifers divergent in genetic merit for fertility traits differ in the age of puberty and reproductive performance. New Zealand's fertility breeding value (FertBV) is the proportion of a sire's daughters expected to calve in the first 42 d of the seasonal calving period. We used the New Zealand national dairy database to identify and select Holstein-Friesian dams with either positive (POS, +5 FertBV, n = 1,334) or negative FertBV (NEG, -5% FertBV, n = 1,662) for insemination with semen from POS or NEG FertBV sires, respectively. The resulting POS and NEG heifers were predicted to have a difference in average FertBV of 10 percentage points. We enrolled 640 heifer calves (POS, n = 324; NEG, n = 316) at 9 d ± 5.4 d (± standard deviation; SD) for the POS calves and 8 d ± 4.4 d old for the NEG calves. Of these, 275 POS and 248 NEG heifers were DNA parent verified and retained for further study. The average FertBV was +5.0% (SD = 0.74) and -5.1% (SD = 1.36) for POS and NEG groups, respectively. Heifers were reared at 2 successive facilities as follows: (1) calf rearing (enrollment to ∼13 wk of age) and (2) grazier, after 13 wk until 22 mo of age. All heifers wore a collar with an activity sensor to monitor estrus events starting at 8 mo of age, and we collected weekly blood samples when individual heifers reached 190 kg of body weight (BW) to measure plasma progesterone concentrations. Puberty was characterized by plasma progesterone concentrations >1 ng/mL in at least 2 of 3 successive weeks. Date of puberty was defined when the first of these samples was >1 ng/mL. Heifers were seasonally bred for 98 d starting at ∼14 mo of age. Transrectal ultrasound was used to confirm pregnancy and combined with activity data to estimate breeding and pregnancy dates. We measured BW every 2 wk, and body condition and stature at 6, 9, 12, and 15 mo of age. The significant FertBV by day interaction for BW was such that the NEG heifers had increasingly greater BW with age. This difference was mirrored with the significant FertBV by month interaction for average daily gain, with the NEG heifers having a greater average daily gain between 9 and 18 mo of age. There was no difference in heifer stature between the POS and NEG heifers. The POS heifers were younger and lighter at puberty, and were at a lesser mature BW, compared with the NEG heifers. As a result, 94 ± 1.6% of the POS and 82 ± 3.2% of the NEG heifers had reached puberty at the start of breeding. The POS heifers were 20% and 11% more likely to be pregnant after 21 d and 42 d of breeding than NEG heifers (relative risk = 1.20, 95% confidence interval of 1.03-1.34; relative risk = 1.11, 95% confidence interval of 1.01-1.16). Results from this experiment support an association between extremes in genetic merit for fertility base on cow traits and heifer reproduction. Our results indicate that heifer puberty and pregnancy rates are affected by genetic merit for fertility traits, and these may be useful phenotypes for genetic selection.

Efficacy of Broilers as a Method of Face Fly (Musca autumnalis De Geer) Larva Control for Organic Dairy Production.

Simple SummaryDairy cows housed on pasture are commonly afflicted by face flies, which develop as larvae in cow manure. Some organic dairy producers graze chickens in succession behind cattle as an effort to disrupt the development of dung fly larvae, but this method has yet to be validated as a successful method of pest management. In a controlled research trial, pastured broiler chickens did not reduce the survival of face fly larvae that developed in cow dung pats. This finding was buoyed by broiler behavioral observations, which revealed the broilers were unwilling to forage in dung pats. In general, broilers were reluctant to leave their shelter as solar radiation increased and climatic conditions were associated with the displacement of time allocated to sitting and sleeping behaviors. Dairy producers should take heed when adopting methods of fly control that rely on broiler chickens to willingly forage in cow dung pats especially when climatic conditions are aversive.The objective of this study was to evaluate Freedom-Ranger broiler chickens as a method to control face fly (Musca autumnalis De Geer) larvae in cow dung pats on pasture. Ninety-nine pats in three replicates were inoculated with first-instar larvae and exposed to one of four treatment conditions for 3 to 4 days: (1) an environment-controlled greenhouse (GH); (2) pasture without broilers (NEG); (3) pasture with 25 broilers stocked at a low density of 2.5 m2 of outdoor area per broiler (LOW); and (4) pasture with 25 broilers stocked at a high density of 0.5 m2 of outdoor area per broiler (HIGH). Broiler behaviors and weather conditions were recorded twice daily. Survival rates of larvae (mean, 95% CI) were similar for pats in the NEG (4.4%, 2–9%), LOW (5.6%, 3–11%), and HIGH (3.2%, 2–7%) groups, and was greatest for larvae reared in the GH (54.4%, 36–72%) group compared to all other groups. The proportion of broilers observed pasture ranging was 14.0% (6–28%) but was negatively related to solar radiation. Broilers were never observed foraging in pats. Results indicate that use of broilers may not be an effective method for controlling larvae of dung pat breeding flies.

Predictive Power of Early, Sequential MRD Monitoring in Peripheral Blood and Bone Marrow in Patients with Mantle Cell Lymphoma Following Autologous Stem Cell Transplantation with or without Rituximab Maintenance; Final Results from the LyMa-MRD Project, Conducted on Behalf of the Lysa Group

Predictive Power of Early, Sequential MRD Monitoring in Peripheral Blood and Bone Marrow in Patients with Mantle Cell Lymphoma Following Autologous Stem Cell Transplantation with or without Rituximab Maintenance; Final Results from the LyMa-MRD Project, Conducted on Behalf of the Lysa Group

Intraperitoneal (IP) port cytology after completion of primary therapy for advanced stage ovarian cancer: A novel approach to a “second look”

ObjectiveTo determine whether IP port cytology predicts early recurrence and/or poor prognosis in patients with ovarian cancer who have completed primary therapy. MethodsA prospective study of patients with advanced stage ovarian cancer undergoing IP port removal after debulking followed by IV/IP chemotherapy was performed. Ports were flushed with 10 cc of normal saline into ThinPrep fixative to be analyzed for cytology. Results were correlated with clinical factors and cancer outcomes. Survivals were calculated using Kaplan-Meier curves and compared using log-rank analysis. ResultsEffluent from 53 IP ports was analyzed, and patients were followed for a median of 62 months. Mean age was 58.5, with the majority of patients being white (90%), with stage 3 (62%), serous histology (87%). Seven (13.2%) patients had positive IP cytology. POS and NEG groups were similar with regard to age, BMI, stage, grade, and GOG status. Patients with POS results had increased risk of recurrence HR 3.2 (95%CI 0.4, 28.9), and death HR 6.5 (95%CI 0.7, 58.8), and were more likely to recur before 12 months, 71% vs. 22% (p = 0.007). Compared to NEG, POS conferred a shorter median survival with PFS of 32 vs. 7 months (p = 0.02) and OS of 84 vs. 42 months (p = 0.04). ConclusionsIP port cytology is predictive of recurrence and survival in patients with ovarian cancer. This inexpensive test may serve as an adjunct to imaging and tumor markers to determine disease status at the completion of treatment. Further study should investigate how this may impact management.

Hydroxymethylnitrofurazone treatment in indeterminate form of chronic Chagas disease: Reduced intensity of tissue parasitism and inflammation-A histopathological study.

Hydroxymethylnitrofurazone (NFOH) is a nitrofurazone prodrug effective invivo during acute infections, and it has less hepatotoxicity effect than the standard drug benznidazole (BZN) which has been used during short- and long-term treatment. In the present study, we induced the indeterminate form of Chagas disease in mice with a Y strain of Trypanosoma cruzi and analysed the histopathological data about the effects of NFOH and BZN on different tissues, including the heart, skeletal muscle, liver, kidney, colon, spleen and brain. After infection, BALB/c mice were treated with NFOH (150mg/kg) and BZN (60mg/kg) for 60days and then submitted to immunosuppression using dexamethasone (5mg/kg) for 14days. Two trained analysts, as part of a blind evaluation, examined the results using serial sections of 3mm diameter in two different moments. The results showed reactivation of the disease only in the infected nontreated group (POS). After treatment, amastigote nests were found in the heart, colon, liver and skeletal muscle in the POS group and in the heart and liver of the BZN group. Interestingly, amastigote nests were not found in the NFOH and NEG groups. The histopathological analysis showed fewer tissue lesions and parasite infiltrates in the NFOH group when compared with the BZN and POS groups. We have not observed any increase in the levels of hepatocellular injury biomarkers (AST/ALT) in the NFOH group. These invivo studies show the potential for NFOH as an effective and safe compound useful as an anti-T.cruzi agent.

Humoral immune responses and viral shedding following vaccination with modified live porcine reproductive and respiratory syndrome virus vaccines

The antibody response and pattern of shedding of vaccine virus following vaccination with modified live genotype I or II porcine reproductive and respiratory syndrome virus (PRRSV) vaccines (MLVs) were investigated. Ninety PRRSV-free pigs were divided randomly seven, groups including the NEG, EU1, EU2, US1, US2, US3 and US4 groups. The NEG group was unvaccinated. The EU1, EU2, US1, US2, US3 and US4 groups were vaccinated with the following MLVs: AMERVAC® PRRS, Porcillis® PRRS, Fostera™ PRRS, Ingelvac® PRRS MLV, Ingelvac® PRRS ATP, and PrimePac™ PRRS+, respectively. Sera were quantitatively assayed for viral RNA using qPCR. Antibody responses were measured using Idexx ELISA and serum neutralization (SN). Shedding of vaccine virus was investigated using sentinel pigs and by detection of viral RNA in tonsil scrapings. Antibody responses were detected by ELISA at 7-14days post-vaccination (DPV) and persisted at high titers until 84 DPV in all MLV groups. The SN titers were delayed and isolate-specific. SN titers were higher for the homologous virus than for heterologous viruses. Age-matched sentinel pigs introduced into the EU2, US2 and US3 groups at 60 DPV seroconverted. In contrast, sentinel pigs introduced at 84 DPV remained negative in all of the MLV groups. Vaccine viral RNA was detected in tonsil scrapings from the EU2, US2 and US3 groups at 84-90 DPV. No viral RNA was detected beyond 70 DPV in the EU1, US1 and US4 groups. In conclusion, all MLV genotypes induced rapid antibody responses, which were measured using ELISA. The development of SN antibodies was delayed and isolate-specific. However, the shedding pattern was variable and depended on the by virus isolate used to manufacture the vaccine.

Antibody-Mediated Rejection in Lung Transplantation: Clinical Outcomes and Donor-Specific Antibody Characteristics.

In the context of lung transplant (LT), because of diagnostic difficulties, antibody-mediated rejection (AMR) remains a matter of debate. We retrospectively analyzed an LT cohort at Foch Hospital to demonstrate the impact of AMR on LT prognosis. AMR diagnosis requires association of clinical symptoms, donor-specific antibodies (DSAs), and C4d(+) staining and/or histological patterns consistent with AMR. Prospective categorization split patients into four groups: (i) DSA positive, AMR positive (DSA(pos) AMR(pos) ); (ii) DSA positive, AMR negative (DSA(pos) AMR(neg) ); (iii) DSA limited, AMR negative (DSA(Lim) ; equal to one specificity, with mean fluorescence intensity of 500-1000 once); and (iv) DSA negative, AMR negative (DSA(neg) ). AMR treatment consisted of a combination of plasmapheresis, intravenous immunoglobulin and rituximab. Among 206 transplanted patients, 10.7% were DSA(pos) AMR(pos) (n = 22), 40.3% were DSA(pos) AMR(neg) (n = 84), 6% were DSA(Lim) (n = 13) and 43% were DSA(neg) (n = 88). Analysis of acute cellular rejection at month 12 showed higher cumulative numbers (mean plus or minus standard deviation) in the DSA(pos) AMR(pos) group (2.1 ± 1.7) compared with DSA(pos) AMR(neg) (1 ± 1.2), DSA(Lim) (0.75 ± 1), and DSA(neg) (0.7 ± 1.23) groups. Multivariate analysis demonstrated AMR as a risk factor for chronic lung allograft dysfunction (hazard ratio [HR] 8.7) and graft loss (HR 7.56) for DSA(pos) AMR(pos) patients. Our results show a negative impact of AMR on LT clinical course and advocate for an early active diagnostic approach and evaluation of therapeutic strategies to improve prognosis.

Characterization of Mannheimia haemolytica in beef calves via nasopharyngeal culture and pulsed-field gel electrophoresis.

Mannheimia haemolytica is a major bacterial component of bovine respiratory disease (BRD); unfortunately, very little is known about M. haemolytica transmission dynamics among cattle. Identifying potential variation in M. haemolytica populations over time and induction of nasopharyngeal colonization and subsequent shedding are 2 areas where knowledge is lacking. In our study, 2 separate loads of 20 mixed-origin, male calves were purchased through an order buyer on different dates. Deep nasopharyngeal cultures (NPC) were performed on all calves on arrival and, if M. haemolytica-negative, a second screening culture was obtained. Calves that were negative on 2 initial NPCs (NEG; n = 4) were subsequently challenged with a previously isolated field strain of M. haemolytica in both the upper and lower respiratory tract, individually housed, and then monitored for M. haemolytica shedding via NPCs at 0.5, 1, 3, 5, 7, and 9 days postchallenge. Naturally M. haemolytica-positive calves (2 per load) were kept for additional daily cultures (POS; n = 4). Individual calf M. haemolytica status for both the POS and NEG groups was inconsistent between study days. Additionally, pulsed-field gel electrophoresis performed on isolates from the positive cultures showed that the NEG calves did not shed the M. haemolytica challenge strain, but rather 2 distinct clusters of M. haemolytica were shared among POS and NEG calves regardless of their initial status. Although sample sizes were small, these findings illustrate how variable the results of a single nasopharyngeal swab can be and the challenges of using an individual culture to truly represent animal M. haemolytica status.

Transfusion Of Thawed Rare Red Blood Cell Units In Finland

The Finnish Red Cross Blood Service performs blood grouping of all blood donations in Finland and serves as a national reference laboratory in pre-transfusion testing of patients. In addition to ABO and RhD blood groups, other Rh antigens and K antigen are also determined from all red cell units. The antigens of JK, FY, and MNS blood group systems are analyzed from selected donors. The extensive donor typing procedure includes antigens from KEL, LW, LU, CO, DO, DI, YT, GE and CROM blood group systems, performed either serologically or by genotyping. Patients negative for a high frequency blood group antigen present a challenge for transfusion laboratories. A blood group is considered rare if the prevalence is 1:1000 or less, with the most common rare phenotypes in Finland being LWa neg, Jk:-3 and Pk. To ensure the availability of matching blood for patients with a rare blood group, we started a freezing program of rare blood in Finland with Haemonetics ACP 215 process in April 2010. The system is closed, therefore after thawing the units are safe for use for 7 days. 18 blood groups which were included in the program were determined based on the known rare blood groups in the Finnish population. Blood groups which are globally rare but more common in Finland, such as Jk:-3 and LWa neg, are represented in our storage and also available internationally. The aim of the present study was to analyze the status of the freezing program of rare red blood cells in Finland. For the analysis we asked the transfusion data of rare red blood cell units delivered to 10 national and 2 international hospitals. Results: In 2010-2012, altogether 204 units of rare blood were stored, including units of all the predefined rare phenotypes except Vel neg. With the exception of Vel neg, Hrs neg and Oh blood groups, we have been able to meet the need for rare blood in Finland using Finnish donors. Altogether 55 units have been thawed and distributed to 27 adult patients. The indication was delivery in 3 cases, surgery in 6 cases, and chronic anemia in 4 cases. For the present analyses the transfusion data of 49 units was available. The data of 6 Coa neg units is missing. Altogether 22 / 49 (45%) of the units were transfused. In 14 cases the blood was transfused to the patient it was intended for, and in 8 cases to another patient, 27 units were discarded. There were no transfusion associated complications. The hemoglobin response was evaluable in 7 patients. In 4 patients with a chronic anemia the median blood hemoglobin response per a unit was +8 g/l, range +2 - +16 g/l. In 3 patients with blood loss during an operation the median increase in blood hemoglobin value was +7 g/l per unit, range +4 – +12 g/l. Conclusions: Compared to the common 24 hour eligibility of thawed red cell units, our units valid for 7 days are preferable in countries like Finland with long distances and in international rare blood deliveries. We have been able to offer rare blood to Finnish patients. The hemoglobin response was reasonable and there were no transfusion associated complications. Our next challenge will be to recruit Finnish blood donors with different ethnic backgrounds (eg. immigrants) and identify their rare blood groups. We have already expanded our program to include rare combinations of common blood groups. Disclosures: No relevant conflicts of interest to declare.

Secretion of prostaglandins and leukotrienes by endometrial cells in cows with subclinical and clinical endometritis

The aims of this study were (1) to measure the secretion of prostaglandin F2α (PGF2α), prostaglandin E2 (PGE2), leukotriene B4 (LTB4), and leukotriene C4 (LTC4) by endometrial cells collected by a cytobrush from healthy cows and cows with subclinical and clinical endometritis in the fourth week postpartum, and (2) to evaluate the relationship between the mediators' levels of secretion and the number of polymorphonuclear neutrophils (PMNs) in the uterine smears of cows with subclinical endometritis. The study included cows without any signs of clinical endometritis (n = 63) and cows with clinical endometritis as a positive control (ENDOM, n = 12). Two different threshold ratios (>5% and >18% of PMNs) were used to categorize the cows without clinical signs as with or without cytologic endometritis (CE). Considering the first or second threshold, the animals with CE were included in group CE POS I or CE POS II, whereas the healthy cows were assigned to group CE NEG I or CE NEG II, respectively.The prevalence of CE was 68.25% (42/63) and 57.14% (36/63) according to the first and second thresholds, respectively. The highest level of secretion of all of the measured mediators occurred in the ENDOM group and differed significantly (P < 0.05) from the CE POS and CE NEG groups, regardless of the threshold. PGF2α secretion in the CE POS II group (1629 pg/mL) was significantly lower (P < 0.05) when compared with the CE NEG II group (2797 pg/mL), whereas there was no significant difference between the CE POS I and CE NEG I groups. PGE2 secretion differed between both groups with CE; higher concentrations were measured in the CE POS II group (6.68 ng/mL) when compared with the CE POS I (2.4 ng/mL) and CE NEG II (2.37 ng/mL) groups (P < 0.05). No significant differences were observed in the LTB4 and LTC4 secretion between the CE POS and CE NEG groups, considering both thresholds. It seems that CE does not fully mimic the inflammatory cascade associated with clinical signs. The response in the subclinical cases was limited to enhanced production of PGE2, which was particularly well-pronounced in cows with high numbers of PMNs (>18%) in the endometrial scrapings.

Natural infection of human adenovirus 36 in rhesus monkeys is associated with gain in body fat, yet, reduction in fasting glucose

Experimental infection of Ad36 increases adiposity, yet improves glycemic control in rodents. In humans, natural Ad36 infection is temporally associated with gain in adiposity and better glycemic control. We determined associations of natural Ad36 infection with changes in adiposity and glycemic control in rhesus monkeys. Monkeys randomized 1y earlier to receive a diet of high fat and sucrose (HFS, N=10) or HFS+resveratrol (RES, N=10), were screened for neutralizing antibodies to Ad36. Ad36‐seropositive (POS; N=5 HFS; 5 RES) and seronegatives (NEG) were compared cross‐sectionally and after 1y follow‐up. In pooled HFS and RES groups, cross‐sectionally, NEG and POS groups did not differ in age, body fat, or fasting glucose (p&gt;;0.9). However, 1y later, fat gain was 510g vs 1245g and reductions in fasting glucose were −3 and −10mg/dL for NEG vs POS, respectively. Gain in body fat and drop in fasting glucose after 1y follow‐up, were significant for POS (p&lt;0.003), but not for NEG. When analyzed separately by diet and seropositivity, only the RES treated POS group showed significant reductions in fasting glucose during 1y follow up (p=0.007). In summary, Ad36 infection is linked with changes in adiposity and glycemic control. Considering effect of resveratrol on reduction in fasting glucose was augmented by Ad36 infection, the interaction may be further evaluated, particularly when studying metabolic interventions.

2108 PREOPERATIVE URINARY EXOSOME DETECTION OF TMPRSS2:ERG MRNA CLOSELY CORRELATES WITH TISSUE EXPRESSION AT RADICAL PROSTATECTOMY

You have accessJournal of UrologyProstate Cancer: Markers I1 Apr 20122108 PREOPERATIVE URINARY EXOSOME DETECTION OF TMPRSS2:ERG MRNA CLOSELY CORRELATES WITH TISSUE EXPRESSION AT RADICAL PROSTATECTOMY Piruz Motamedinia, Anna Scott, Kendall Bate, Guillermo Salazar, Michael Lipsky, Neda Sadeghi, Wayne Comper, James McKiernan, and Leileata Russo Piruz MotamediniaPiruz Motamedinia New York, NY More articles by this author , Anna ScottAnna Scott New York, NY More articles by this author , Kendall BateKendall Bate New York, NY More articles by this author , Guillermo SalazarGuillermo Salazar New York, NY More articles by this author , Michael LipskyMichael Lipsky New York, NY More articles by this author , Neda SadeghiNeda Sadeghi New York, NY More articles by this author , Wayne ComperWayne Comper New York, NY More articles by this author , James McKiernanJames McKiernan New York, NY More articles by this author , and Leileata RussoLeileata Russo New York, NY More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.2276AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Exosomes (including microvesicles) are lipid bilayer vesicles that are released into biofluids such as urine and carry high integrity RNA from the parent cell. We have recently shown that urinary exosomes can be isolated and found to contain prostate specific mRNA transcripts. Here we examine the concordance of our detection of TMPRSS2:ERG (T:E) in urinary exosomes versus prostatectomy tissue and determine the rate of T:E detection in various prostate cancer related groups. METHODS Following Columbia University IRB approval, random urine samples (without prostate massage) were collected from 163 men who were stratified into 4 groups: TRUS prostate biopsy negative (Bx Neg, n=39), TRUS biopsy positive (Bx Pos, n=47), post-radical prostatectomy (RP, n=37) and controls (males <35 yrs, n=40). Additionally, whole-mount paraffin embedded prostate sections were obtained from 11 patients who underwent RP and had pre-RP urine specimens available (7 T:E positive, 4 T:E negative in pre-RP urine). Urine samples were stored at 4°C and 0.8 μm filtration was used to remove whole cells and debris. Urinary exosomal RNA and tissue RNA were isolated using an in-house technique (Exosome Diagnostics, NY) and analyzed using RP-qPCR. RESULTS Mean serum PSA levels were similar in the Bx Neg and Bx Pos groups. All groups except the control (males <35 yrs) were age matched. T:E fusion events were found to occur in 68% of Bx Pos and 44% of Bx Neg patients. Only 5% of controls expressed T:E and no patient in the post RP group had positive T:E detection. Furthermore, T:E expression was lost in the 4 Bx Pos patients that underwent prostatectomy and were retested following surgery. Tissue analysis revealed positive T:E in 8 of 11 prostates. T:E expression concordance between urine and tissue occurred in 10 of 11 (91%) patients with a sensitivity and specificity of 89% and 100%, respectively. CONCLUSIONS The detection rate of the T:E fusion in the Bx Pos group using urinary exosomal RNA was consistent with previous reports using tissue analyses. Our study exhibited near perfect concordance between urinary exosomal mRNA with matched prostate tissue analysis. The detection of T:E expression in the urine of patients with a negative TRUS biopsy may be due to false negative biopsy results or assay detection of T:E expression in premalignant lesions. The unique stability and yield of urinary exosomal RNA will broaden the role of exosomes in future diagnostic testing and simplify sample handing without the variability and patient discomfort inherent to prostate massage. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e850-e851 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Piruz Motamedinia New York, NY More articles by this author Anna Scott New York, NY More articles by this author Kendall Bate New York, NY More articles by this author Guillermo Salazar New York, NY More articles by this author Michael Lipsky New York, NY More articles by this author Neda Sadeghi New York, NY More articles by this author Wayne Comper New York, NY More articles by this author James McKiernan New York, NY More articles by this author Leileata Russo New York, NY More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Urinary exosomes as a stable source of mRNA for prostate cancer analysis.

174 Background: Exosomes are novel lipid bilayer vesicles that are released into biofluids such as urine and carry high integrity RNA from the parent cell which they were derived. Due to their unique stability and the fact that they contain prostate specific mRNA transcripts, we examined their potential as a non-invasive source of mRNA biomarkers for prostate cancer analysis. Methods: Following a Columbia University approved IRB protocol, random urine samples were collected from 163 men who were stratified into 4 groups: biopsy negative (Bx Neg, n=39), biopsy positive (Bx Pos, n=47), post-radical prostatectomy (RP, n=37) and controls (males &lt;35 yrs, n=40). Urine samples were stored at 4°C and 0.8 μm filtration was used to remove whole cells and debris. Urinary exosomal RNA was isolated using our in-house technique. RT-qPCR was used to analyze PSA, PCA3, androgen receptor (AR), survivin, NCOA2, RAD21, transmembrane protease serine 2 (TMPRSS2), ERG and TMPRSS2:ERG fusion at the mRNA level. Results: Mean serum PSA protein level and age were similar in the Bx Neg and Bx Pos groups. Relative quantitation (RQ) of genes standardized to the PSA gene revealed that ERG (P&lt;0.005), PCA3 (P&lt;0.005), TMPRSS2:ERG fusion (P&lt;0.05), TMPRSS2 (P&lt;0.05) and survivin (P&lt;0.005) were significantly increased in the Bx Pos vs. Bx Neg group. TMPRSS2:ERG fusion events occurred in 68% of Bx Pos vs. 44% of Bx Neg patients and were present in only 5% of controls. No patient in the RP group had positive TMPRSS2:ERG detection; this finding was further supported by the loss of TMPRSS2:ERG expression for 4 Bx Pos patients following prostatectomy suggesting specificity of the fusion event to the prostate. Conclusions: This study confirms urinary exosomes as a source of high quality RNA and showed significant differences in the expression of ERG, PCA3, TMPRSS2:ERG genes between the Bx Pos and Bx Neg groups. Our findings are consistent with previous studies based on tissue and post-prostate massage urinary cell analyses. The unique stability and yield of urinary exosomal RNA collected without a prostatic massage will hopefully simplify sample handing, obviate sample variability and patient discomfort inherent to prostate massage, and broaden the role of exosomes in future diagnostic testing.

Abstract C133: The use of exosomal mRNA for noninvasive transcriptional analysis of the prostate.

Abstract Background: Prostate cancer studies have used urine as a non-invasive source of nucleic acids for biomarker analysis. These studies have been limited by the need for a digital rectal exam (DRE) or prostate massage prior to urine collection to enable enough cellular material for RNA analysis. Such manipulations can potentially lead to variability in assay performance depending on DRE intensity and may also reduce compliance. Here we investigate the novel use of RNA in urinary exosomes, small lipid bilayer vesicles released from cells into bodily fluids, to analyze previously identified prostate cancer mRNA biomarkers without a prostate massage. Methods: Random urine samples (&amp;gt;20ml) were collected from 207 consecutive patients under a Columbia University IRB approved prospective protocol. Patients were stratified into 5 groups: TRUS biopsy negative (Bx Neg, n=39), TRUS biopsy positive (Bx Pos, n=47), post-radical prostatectomy (RP, n=37), no TRUS biopsy (No Bx, n=44) and control (healthy males &amp;lt;35 yrs, n=40). Due to the unique stability of exosomal RNA (exoRNA), urine samples were stored at 4°C and 0.8 um filtration was used to remove whole cells and debris. Urinary exoRNA was isolated using our in-house technique. RT-qPCR was used to analyze GAPDH, prostate-specific antigen (PSA), prostate cancer antigen 3 (PCA3), androgen receptor (AR), survivin, nuclear receptor co-activator 2 (NCOA2), RAD21, transmembrane protease serine 2 (TMPRSS2), ERG and TMPRSS2:ERG fusion. Results: All groups were age matched except for control males (&amp;lt;35 yrs)(P&amp;lt;0.001). Serum PSA protein was significantly lower in the No Bx and RP groups versus the Bx Neg and Bx Pos groups (P&amp;lt;0.001). Relative quantitation (RQ) of genes standardized to GAPDH revealed that ERG and PCA3 expression levels were significantly increased in the Bx Pos versus Bx Neg groups (ERG P&amp;lt;0.01, PCA3 P&amp;lt;0.05). Further, RQ analysis standardized to the prostate related gene, PSA, revealed that survivin (P&amp;lt;0.005), ERG (P&amp;lt;0.005), PCA3 (P&amp;lt;0.005), TMPRSS2:ERG fusion (P&amp;lt;0.05), and TMPRSS2 (P&amp;lt;0.05) were significantly increased in the Bx Pos versus Bx Neg group. Analysis of the TMPRSS2:ERG fusion revealed 44% of Bx Neg and 68% of Bx Pos had a positive fusion event. Analysis of the RP group revealed that no patients had positive TMPRSS2:ERG detection; this finding was further supported by the loss of TMPRSS2:ERG expression experienced by 4 Bx Pos patients following prostatectomy suggesting specificity of the fusion event for the prostate. Interestingly, the control group (males &amp;lt;35yrs) had a much lower TMPRSS2:ERG fusion incidence of 5% compared to the No Bx group's fusion incidence of 41%, which was similar to the Bx Neg group despite having significantly lower serum PSA protein levels. Conclusion: This study confirms that urinary exosomes are a novel source of high quality RNA to examine prostate gene expression. We demonstrate that expression levels of ERG, PCA3, TMPRSS2:ERG, TMPRSS2 and survivin are significantly higher in cases of confirmed prostate cancer, consistent with previous studies using prostate biopsies and/or urinary cells collected after DRE/prostate massage. The unique stability and yield of urinary exoRNA collected without the need for a prostatic massage will hopefully simplify sample handing, obviate sample variability and patient discomfort inherent to prostate massage, and broaden the role of exosomes in future diagnostic testing. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr C133.

P10 High troponin I in acute liver failure: a marker of myocardial injury or metabolic stress?

IntroductionALF is a life-threatening multi-system illness resulting from massive hepatic necrosis. Acute liver failure (ALF), in its more severe forms is invariably complicated by progressive haemodynamic disturbances with a pattern...

The Prevalence of Gastroesophageal Reflux in Patients with Paradoxical Vocal Fold Motion

Purpose: Paradoxical vocal fold motion (PVFM) is an inappropriate adduction of the true vocal folds, which causes airway obstruction and inspiratory stridor. PVFM may lead to wheezing and is often mistaken for asthma, and patients are treated with glucocorticoids and beta-agonists inappropriately. There is a body of evidence stating that irritation of the vocal folds may cause or exacerbate PVFM. One-third of PVFM patients have been shown to report GERD symptoms, but studies to evaluate PVFM patients with pH monitoring have all been small (<10 subjects). We studied patients with PVFM to determine the prevalence of GERD in this population as diagnosed by esophagogastroduodenoscopy (EGD) or 48-hour esophageal acid monitoring. Methods: Patients diagnosed with PVFM by flattened inspiratory loop on pulmonary function testing underwent EGD with BRAVO capsule placement at 6 cm proximal to SCJ. All patients abstained from acid suppression agents (PPI's, H2RA's) for 7 days prior to the EGD. 48 hours of pH monitoring was performed with the BRAVO capsule. Patients were considered to have a diagnosis of GERD if they had any evidence of erosive esophagitis on EGD or if they had a positive Johnston-Demeester (JD) score (>22) during the second 24-hour period of pH monitoring by BRAVO capsule. Results: 32 patients (20 females, mean age 42 + 16 yrs) with PVFM completed the study. 31% (10/32) of subjects had a positive JD score, 50% of which had significant combined upright and supine reflux. On EGD, 18.8% (6/32) had evidence of esophagitis. Two subjects with esophagitis had a normal JD score. In this study the prevalence of GERD in PVFM was 37.5% (12/32, 95% CI 21.1–56.3%). Of GERD positive subjects, 50% (6/12) had evidence of esophagitis and 50% had atypical or no symptoms of GERD. GERD pos and GERD neg groups were similar in mean age (43 + 16 yrs vs. 41 + 16 yrs, P= 0.708) and sex (female, 50% of GERD pos vs. 70% of GERD neg, P= 0.452). Conclusion: GERD prevalence in this study was 37.5%. This suggests a high prevalence of GERD in the PVFM population. Because of this, PVFM patients should be studied for GERD, especially since many are asymptomatic. This may also suggest that GERD plays a role in the pathogenesis of PVFM.

Neomycin prevents enzyme-mediated glycosaminoglycan degradation in bioprosthetic heart valves

Bioprosthetic heart valves (BHVs) derived from glutaraldehyde crosslinked porcine aortic valves are frequently used in heart valve replacement surgeries. However, BHVs have limited durability and fail either due to degeneration or calcification. Glycosaminoglycans (GAGs), one of the integral components of heart valve cuspal tissue, are not stabilized by conventional glutaraldehyde crosslinking. Previously we have shown that valvular GAGs could be chemically fixed with GAG-targeted chemistry. However, chemically stabilized GAGs were only partially stable to enzymatic degradation. In the present study an enzyme inhibitor was incorporated in the cusps to effectively prevent enzymatic degradation. Thus, neomycin trisulfate, a known hyaluronidase inhibitor, was incorporated in cusps via 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/ N-hydroxysuccinimide (EDC/NHS) chemistry followed by glutaraldehyde crosslinking (NEG). Controls included cusps crosslinked with either EDC/NHS followed by glutaraldehyde (ENG) or only with glutaraldehyde (GLUT). NEG group showed improved resistance to in vitro enzymatic degradation as compared to GLUT and ENG groups. All groups showed similar collagen stability, measured as a thermal denaturation temperature by differential scanning calorimetry (DSC). The cusps were implanted subdermally in rats to study in vivo degradation of GAGs. NEG group preserved significantly more GAGs than ENG and GLUT. NEG and ENG groups showed reduced calcification than GLUT.