Two-tailed Student Research Articles

What Is the Effect of Vitamin C on Finger Stiffness After Distal Radius Fracture? A Double-blind, Placebo-controlled Randomized Trial.

It is proposed that vitamin C administration can reduce disproportionate pain and stiffness after distal radius fracture; however, randomized trials that tested this hypothesis have had inconsistent results. (1) Is administering vitamin C after distal radius fracture associated with better ROM, patient-reported upper extremity function, and pain scores? (2) What factors are associated with post-fracture finger stiffness and worse upper extremity function? This is a double-blind, randomized, placebo-controlled, noncrossover study. Between August 2014 and July 2017, we approached 204 consecutive patients, of which 195 were eligible, and 134 chose to participate. Participants were randomized to receive once-daily 500 mg vitamin C (67 participants) or placebo (67 participants) within 2 weeks after distal radius fracture. All patients received usual care at the discretion of their surgeon. The mean age of participants was 49 ± 17 years, 99 patients (74%) were women, and 83 (62%) were treated nonoperatively. The primary outcome was the distance between the fingertip and distal palmar crease 6 weeks after fracture. This measure is easy to obtain and previously has been shown to correlate with aggregate ROM of all finger joints. The secondary outcomes were total active finger motion, total active thumb motion, upper extremity-specific limitations, and pain intensity.An a priori power analysis suggested 126 patients would provide 80% power to detect a difference of 2 cm (SD 4.0) fingertip distance to palmar crease with α set at 0.05 using a two-tailed Student's t-test. Accounting for 5% lost to followup, we included 134 patients.All analyses were intention-to-treat. Ten participants of the intervention group and five of the placebo group were lost to followup. Their missing data were addressed by multiple imputation, after which we performed linear regression analysis for our outcome variables. Administration of vitamin C was not associated with ROM, function, or pain scores at 6 weeks (distance to palmar crease: β -0.23; 95% CI -1.7 to 1.2; p = 0.754; finger ROM: β 4.9; 95% CI, -40 to 50; p = 0.829; thumb ROM: β 0.98; 95% CI, -18 to 20; p = 0.918, Patient-Reported Outcomes Measurement Information System [PROMIS] score: β 0.32; 95% CI, -2.6 to 3.2; p = 0.828; pain score: β -0.62; 95% CI, -0.62 to 0.89; p = 0.729) nor at 6 months (PROMIS score: β -0.21; 95% CI, -3.7 to 3.3; p = 0.904; pain score: β 0.31; 95% CI, -0.74 to 1.4; p = 0.559). At 6 weeks, we found that more finger stiffness was mildly associated with greater age (β -1.5; 95% CI, -2.8 to -0.083; p = 0.038). Thumb stiffness was mildly associated with greater age (β -0.72; 95% CI, -1.3 to -0.18; p = 0.009) and strongly associated with operative treatment (β -32; 95% CI, -50 to -13; p = 0.001). Greater pain interference was modestly associated with greater functional limitations at 6 weeks (β -0.32; 95% CI, -0.52 to -0.12; p = 0.002) and 6 months (β -0.36; 95% CI, -0.60 to -0.11; p = 0.004). Vitamin C does not seem to facilitate recovery after distal radius fracture, but amelioration of maladaptation to nociception (pain interference) merits greater attention. Level I, therapeutic study.

CVEMP correlated with imbalance in a mouse model of vestibular disorder

BackgroundCervical vestibular evoked myogenic potential (cVEMP) testing is a strong tool that enables objective determination of balance functions in humans. However, it remains unknown whether cVEMP correctly expresses vestibular disorder in mice.ObjectiveIn this study, correlations of cVEMP with scores for balance-related behavior tests including rotarod, beam, and air-righting reflex tests were determined in ICR mice with vestibular disorder induced by 3,3′-iminodipropiontrile (IDPN) as a mouse model of vestibular disorder.MethodsMale ICR mice at 4 weeks of age were orally administered IDPN in saline (28 mmol/kg body weight) once. Rotarod, beam crossing, and air-righting reflex tests were performed before and 3–4 days after oral exposure one time to IDPN to determine balance functions. The saccule and utricles were labeled with fluorescein phalloidin. cVEMP measurements were performed for mice in the control and IDPN groups. Finally, the correlations between the scores of behavior tests and the amplitude or latency of cVEMP were determined with Spearman’s rank correlation coefficient. Two-tailed Student’s t test and Welch’s t test were used to determine a significant difference between the two groups. A difference with p < 0.05 was considered to indicate statistical significance.ResultsAfter oral administration of IDPN at 28 mmol/kg, scores of the rotarod, beam, and air-righting reflex tests in the IDPN group were significantly lower than those in the control group. The numbers of hair cells in the saccule, utricle, and cupula were decreased in the IDPN group. cVEMP in the IDPN group was significantly decreased in amplitude and increased in latency compared to those in the control group. cVEMP amplitude had significant correlations with the numbers of hair cells as well as scores for all of the behavior tests in mice.ConclusionsThis study demonstrated impaired cVEMP and correlations of cVEMP with imbalance determined by behavior tests in a mouse model of vestibular disorder.

Patient-Derived Non-Muscular Invasive Bladder Cancer Xenografts of Main Molecular Subtypes of the Tumor for Anti-Pd-l1 Treatment Assessment.

Background: Establishment of heterotopic patient-derived xenografts of primary and relapsed non-muscular invasive bladder cancer (NMIBC) to explore the biological property of PD-L1 signaling that may impact bladder tumor growth in humanized animals. Methods: Tumor cells of luminal, basal, and p53 subtypes of primary and relapsed NMIBC were engrafted to irradiated (3.5 Gy) NOG/SCID female mice along with intraperitoneal transplantation of human lymphocytes (5 × 107 cells/mouse); a role of PD-L1 signaling pathway inhibition for bladder cancer growth was assessed in humanized animals that carried PD-L1-expressing main molecular subtypes of bladder carcinoma patient-derived xenografts (PDX) and provided with selective anti-PD-L1 treatment. We used two-tailed Student’s t test to explore differences between main and control subgroups. Significance of intergroup comparison was measured with one-way ANOVA followed by the Tukey’s or Newman–Keul’s criterion. Survival curves were analyzed with the Gehan’s criterion with the Yate’s correction. The Spearman’s correlation was used to assess the link between CD8+ expression and sPD-L1 serum level. Differences were considered statistically significant at p < 0.05. Results: Heterotopic primary and relapsed luminal, basal, and p53 subtypes of NMIBC PDXs were established. More than 25% of counted tumor cells of all PDX specimens expressed PD-L1, so the tumors were ranged as PD-L1 positive. Anti-PD-L1 intervention increased survival of the animals that carried both primary and relapsed luminal noninvasive, muscular invasive, and relapsed luminal bladder cancer xenografts. There was significant retardation of tumor volume duplication time in aforementioned subgroups correlated with PD-L1 expression. Bad response of p53 mutant subtypes of NMIBC on specific anti-PD-L1 treatment may be associated with low CD8+ cells representation into the tumors tissue. Conclusions: Established PD-L1-positive NMIBC PDXs differently replied on anti-PD-L1 treatment due to both NMIBC molecular subtype and tumor T-suppressors population. The results may have major implications for further clinical investigations.

Monitoring of acoustic cavitation in microbubble-presented focused ultrasound exposure using gradient-echo MRI.

Gadolinium-based contrast agents can be used to identify the blood-brain barrier (BBB) opening after inducing a focused ultrasound (FUS) cavitation effect in the presence of microbubbles. However, the use of gadolinium may be limited for frequent routine monitoring of the BBB opening in clinical applications. To use a gradient-echo sequence without contrast agent administration for monitoring of acoustic cavitation. Animal and phantom prospective. Static and flowing gel phantoms; six normal adult male Sprague-Dawley rats. 3T, 7T; fast low-angle shot sequence. Burst FUS with acoustic pressures = 1.5, 2.2, 2.8 MPa; pulse repetition frequencies = 1, 10,100 Hz; and duty cycles = 2%, 5%, 10% were transmitted to the chamber of a static phantom with microbubble concentrations = 10%, 1%, 0.1%. MR slice thicknesses = 3, 6, 8 mm were acquired. In flowing phantom experiments, 0.1%, 0.25%, 0.5%, 0.75%, and 1% microbubbles were infused and transmitted by burst FUS with an acoustic pressure = 0.4 and 1 MPa. In in vivo experiments, 0.25% microbubbles was infused and 0.8 MPa burst FUS was transmitted to targeted brain tissue beneath the superior sagittal sinus. The mean signal intensity (SI) was normalized using the mean SI from pre-FUS. Two-tailed Student's t-test. P < 0.05 was considered statistically significant. In the static phantom, the time courses of normalized SI decreases to minimum SI levels of 70-80%. In the flowing phantom, substantial normalized SI of 160-230% was present with variant acoustic pressures and microbubble concentrations. Compared with in vivo control rats, the brain tissue of experimental rats with transmission of FUS pulses exhibited considerable decreases of normalized SI (P < 0.001) because of the cavitation-induced perturbation of flow. Observing gradient-echo SI changes can help monitor the targeted location of microbubble-enhanced FUS, which in turn assists the monitoring of the BBB opening. 2 Technical Efficacy: Stage 1 J. Magn. Reson. Imaging 2020;51:311-318.

Myocardial native-T1 times are elevated as a function of hypertrophy, HbA1c, and heart rate in diabetic adults without diffuse fibrosis

PurposeCardiac native-T1 times have correlated to extracellular volume fraction in patients with confirmed fibrosis. However, whether other factors that can occur either alongside or independently of fibrosis including increased intracellular water volume, altered magnetization transfer (MT), or glycation of hemoglobin, lengthen T1 times in the absence of fibrosis remains unclear. The current study examined whether native-T1 times are elevated in hypertrophic diabetics with elevated hemoglobin A1C (HbA1c) without diffuse fibrosis. MethodsNative-T1 times were quantified in 27 diabetic and 10 healthy adults using a modified Look-Locker imaging (MOLLI) sequence at 1.5 T. The MT ratio (MTR) was quantified using dual flip angle cine balanced steady-state free precession. Gadolinium (0.2 mmol/kg Gd-DTPA) was administered as a bolus and post-contrast T1-times were quantified after 15 min. Means were compared using a two-tailed student's t-test, while correlations were assessed using Pearson's correlations. ResultsWhile left ventricular volumes, ejection fraction, and cardiac output were similar between groups, left ventricular mass and mass-to-volume ratio (MVR) were significantly higher in diabetic adults. Mean ECV (0.25 ± 0.02 Healthy vs. 0.25 ± 0.03 Diabetic, P = 0.47) and MTR (125 ± 16% Healthy vs. 125 ± 9% Diabetic, P = 0.97) were similar, however native-T1 times were significantly higher in diabetics (1016 ± 21 ms Healthy vs. 1056 ± 31 ms Diabetic, P = 0.00051). Global native-T1 times correlated with MVR (ρ = 0.43, P = 0.008) and plasma HbA1c levels (ρ = 0.43, P = 0.0088) but not ECV (ρ = 0.06, P = 0.73). Septal native-T1 times correlated with septal wall thickness (ρ = 0.50, P = 0.001). ConclusionIn diabetic adults with normal ECV values, elevated native-T1 times may reflect increased intracellular water volume and changes secondary to increased hemoglobin glycation.

The nucleoporin ELYS regulates nuclear size by controlling NPC number and nuclear import capacity.

How intracellular organelles acquire their characteristic sizes is a fundamental question in cell biology. Given stereotypical changes in nuclear size in cancer, it is important to understand the mechanisms that control nuclear size in human cells. Using a high-throughput imaging RNAi screen, we identify and mechanistically characterize ELYS, a nucleoporin required for post-mitotic nuclear pore complex (NPC) assembly, as a determinant of nuclear size in mammalian cells. ELYS knockdown results in small nuclei, reduced nuclear lamin B2 localization, lower NPC density, and decreased nuclear import. Increasing nuclear import by importin α overexpression rescues nuclear size and lamin B2 import, while inhibiting importin α/β-mediated nuclear import decreases nuclear size. Conversely, ELYS overexpression increases nuclear size, enriches nuclear lamin B2 at the nuclear periphery, and elevates NPC density and nuclear import. Consistent with these observations, knockdown or inhibition of exportin 1 increases nuclear size. Thus, we identify ELYS as a novel positive effector of mammalian nuclear size and propose that nuclear size is sensitive to NPC density and nuclear import capacity.

Earlier Endotracheal Intubation by Entropy Guidance Versus Neuromuscular Monitoring

Background: The entropy electrode is centrally placed on the forehead over the muscles of frontalis, orbicularis oculi, and corrugator supercilii. It determines response entropy (RE), which is the electromyogram component, and state entropy (SE), which is the electroencephalogram component. We hypothesized that due to the central location of entropy, the decreasing value of RE-SE ≤ 2 with SE ≤ 45 could denote an adequate combination of hypnosis, muscle paralysis, and analgesia required for endotracheal intubation. This could result in earlier intubation compared to when guided by train-of-four (TOF) = 0. Objectives: The primary objective of the study was to evaluate if entropy values of RE-SE ≤ 2 with SE ≤ 45 can be used as a measure of adequate condition for endotracheal intubation. We also sought to determine the TOF at this point. Methods: Endotracheal intubation was performed in group E (Entropy; n = 30) at RE-SE ≤ 2 with SE ≤ 45 and in group T (TOF; n = 30) at TOF = 0. A propofol bolus (20 mg) was administered if the patient had a hypertensive response or moved in response to endotracheal intubation. The TOF was noted at the time of intubation in group E. We also measured the time to intubation, jaw and vocal cord relaxation, patient movement or coughing, SE, TOF, and vital parameters. Statistical analysis was performed with two-tailed students’ t test, paired t test, chi-square test, and ANOVA. The difference between groups was considered significant if the p value was < 0.05. Results: The time to intubation was significantly shorter in group E than in group T (92.5 ± 63.5 seconds vs. 209.2 ± 59.6 seconds; P < 0.001) with a mean TOF of 87.3% ± 8.4% in group E. Intubating conditions in terms of jaw relaxation, patient movement, and coughing were not significantly different between the two groups. Vocal cord relaxation was significantly inadequate in six patients in group E (P < 0.01); however, there was no difficulty in introducing the endotracheal tube with no postoperative adverse effects such as sore throat. Conclusions: Adequate conditions for endotracheal intubation were achieved 90 seconds after the administration of fentanyl, propofol, and vecuronium for anesthesia induction when it is guided by RE-SE ≤ 2 with SE ≤ 45, which is earlier than when guided by TOF = 0.

MON-218 Inflammatory Role of Sex Steroids in Hidradenitis Suppurativa: An Androgenic Phenotype

Hidradenitis Suppurativa (HS) is an autoinflammatory skin disease more common in women and associated with hyper-androgenic disorders such as polycystic ovary syndrome. HS is characterized by chronic recurrent deep-seated painful nodules and draining tunnels in skin areas that follow the distribution of pubertal terminal hair growth. In women, HS disease activity varies with hormonal fluctuation across the menstrual cycle and pregnancy. Anti-androgens have shown promise in treatment of HS, but the pathophysiologic link between sex hormones and HS is unclear. We sought to further elucidate the role of sex hormones in the pathogenesis of HS. Objective: To determine if there is differential estrogen and androgen regulated gene expression in lesional compared to non-lesional skin transcriptome in HS patients. Methods: (a) We analyzed mRNA microarray data from lesional and non-lesional HS skin (GEO, GSE72702) (Blok JL, Br J Dermatol. 2016;174(6):1392). Inflammatory lesional skin from affected axilla or groin and non-lesional skin from the upper arm or leg was obtained from 17 patients with moderate to severe HS. We then examined differential expression of hormone regulated genes identified in existing androgen and estrogen gene expression signatures from prostate cancer cells [(VCaP treated with dihydrotestosterone (DHT)] and breast cancer cells [MCF7 treated with 17 beta-estradiol, (E2)] in the lesional and non-lesional skin transcriptomes of the HS patients. We identified overlap between the sets of genes that were either both upregulated or both repressed in HS lesions and the hormone-responsive prostate and breast cancer cell line models. Significance of differential gene expression was defined as FDR-adjusted p-value for two-tailed Student t-test less than 0.05. (b) After overlapping genes were identified, we used GeneGo Metacore pathway analysis software to explore the functional significance of the identified genes. Results: (a) Of the 3379 E2-upregulated probe sets in MCF7 breast cancer cells, 647 were significantly upregulated and 584 of 2871 E2-downregulated probe sets were significantly downregulated in HS lesions. Of the 1328 DHT-upregulated probe sets in prostate cancer cells, 341 were significantly upregulated and 196 of the 1251 DHT-downregulated probe sets were significantly downregulated in HS lesions. (b) Initial pathway analysis indicates that DHT upregulated genes in HS lesions are strongly enriched in innate immunity pathways, specifically, type 1 interferon signaling and class I MHC antigen presentation. Conclusion: There is substantial overlap between the transcriptional programme of sex steroids and transcriptional changes observed in HS lesional skin. In particular, androgen target genes are significantly enriched in innate immunity pathways, suggesting a functional link between the androgen signaling and the inflammatory process in HS lesions.

SAT-412 Effects of the Ovary and Dihydrotestosterone on Firing Rate of GnRH Neurons in Prenatally Androgenized Mice

GnRH pulse frequency drives downstream reproductive function by signaling for differential release of gonadotropins, which subsequently control ovarian function. In patients with polycystic-ovary syndrome (PCOS), neuroendocrine defects, including persistent high-frequency LH (and presumably GnRH) pulses, are coupled with dysregulated ovarian function. Prenatally androgenized (PNA) animal models exhibit features similar to PCOS, including increased androgens and high-frequency LH pulses. Adult PNA mice exhibit these phenotypes and also have increased GnRH neuron firing rate and increased excitatory GABAergic transmission to GnRH neurons. In contrast to adults, GnRH neuron firing rate at 3 weeks of age is reduced in PNA females. The causes of this age-dependent difference in firing rate are not known, however puberty occurs between these ages and androgens can increase GnRH neuron firing rate in adulthood. Consistent with this, ovariectomy in adult PNA mice decreased firing rate to that in control diestrous mice. Here we tested the hypothesis that a lack of ovarian androgens mediated this decrease. We also investigated if ovariectomy changed firing rate in 3-week old PNA mice. PNA mice were born from dams injected daily on days 16-18 of gestation with 225µg dihydrotestosterone (DHT). Ovariectomy (OVX) or sham surgery was performed and at this time some mice received constant-release DHT implants that produce sub-male levels of androgen (400 µg/implant) 5-7 days before recordings (surgery at 14-16 days of age for prepubertal mice). Hour-long extracellular recordings were made to ascertain spontaneous firing rate of GFP-identified GnRH neurons in acutely-prepared brain slices. In adult OVX PNA mice, DHT increased firing rate of GnRH neurons relative to OVX PNA, but firing rate was still less than in sham-operated PNA animals (two-way ANOVA/Fisher’s LSD, n= 7-11 cells/group). In adult OVX control mice, preliminary data indicate that DHT replacement alone does not alter firing rate, in contrast to previous results with DHT plus estradiol. At three weeks of age, there was no difference in firing rate between PNA sham and OVX mice (unpaired two-tailed Student’s t-test, n=7-8 cells/group). These studies suggest that increased GnRH firing rate in adult PNA females is driven at least in part by ovarian androgens, whereas changes in firing rate in 3-week-old PNA females are driven by a different mechanism, such as prenatal programming, rather than ovarian factors.

Sequential Interventional Management of Osseous Neoplasms via Embolization, Cryoablation, and Osteoplasty.

The purpose of this study is to determine if sequential interventional therapy can become a mainstay option in providing palliation from fastidious osseous neoplasms in patients with pain refractory to oral analgesia and radiotherapy. This retrospective monocentric study was approved by our institutional review board. Between July 2012 and August 2014, we reviewed 15 patients (6 women, 9 men; age range of 36-81 years) who underwent embolization followed by cryoablation, with or without osteoplasty. Patient demographics and tumor characteristics, including primary histology and the location of metastasis, were included in our review. Pain intensity at baseline, after radiotherapy, and after sequential interventional therapy was reviewed using the hospital electronic medical record. The use of oral analgesia and procedural complications was also noted. Data was then assessed for normality and a two-tailed Student's t-test was performed on mean pain scores for difference phases of treatment. While radiotherapy offers pain relief with a mean pain score of 7.25 ±1.5 (p =<.0001), sequential interventional therapy results in better comfort as demonstrated by a mean pain score of 3.9 ± 2.6 (p=.0015). Moreover, all patients who reported oral analgesic use at presentation reported a decrease in their requirement after sequential interventional therapy. Embolization and cryoablation were performed in all patients, while osteoplasty was indicated in 6 cases. There was no difference in postprocedural pain intensity between patients who required osteoplasty and patients who did not (p = 0.7514). There were no complications observed during treatment. This retrospective study shows that sequential intervention with transarterial embolization, cryoablation, and osteoplasty is both safe and efficacious for bone pain refractory to the current standard of care. We demonstrated that this combination therapy has the potential to become an effective mainstay treatment paradigm in the palliative care of osseous neoplasm to improve quality of life.

Alginate Oligosaccharide Attenuates α2,6--Sialylation Modification to Inhibit Prostate Cancer Cells Growth via Hippo / YAP Pathway

Background: Alginate oligosaccharide (AOS), a water-soluble marine plant oligosaccharide, was obtained by enzymatic hydrolysis of sodium alginate. Many studies have shown chitosan oligosaccharides can inhibit various tumors. Although AOS has been shown to induce apoptosis in osteosarcoma with fewer side effects, the inhibitory effect of AOS on prostate cancer and its molecular mechanism remain obscure. Methods: Luciferase reporter assay was used to detect effect of AOS on transcriptional activity of ST6Gal-1 promoter and identification of core functional regions; Chromatin immunoprecipitation (CHIP) was used to further validate the transcription factor interacting with the core region of ST6Gal-1 promoter; Co-immunoprecipitation (Co-IP) was used to verify the protein interaction between transcription factor c-Jun that was interacted with ST6Gal-1 promoter and YAP. The change of tumor-forming ability after AOS treatment was detected in athymic nude mice in vivo. Statistical significance was estimated by a two-tailed Student’s t-test and ANOVA. Findings: This study showed AOS inhibited cell growth, which was consistent with attenuation of α2,6-sialylation modification. Meanwhile, AOS inhibited activity of ST6Gal-1 promoter and affected transcriptional process. In addition, AOS could activate Hippo/YAP pathway and block the recruitment of coactivator YAP and c-Jun to ST6Gal-1 promoter. Furthermore, YAP interacted with transcription factor c-Jun to regulate transcriptional activity of downstream target ST6Gal-1 gene. Consistent with the in vitro data, AOS suppressed tumorigenicity of prostate cancer cells via Hippo/YAP pathway in vivo. Interpretation: AOS results in slowing proliferation of prostate cancer and can be developed as an anti-tumor adjuvant for treatment of prostate cancer cells. Funding Statement: This work was supported by the National Natural Science Foundation of China (31470799); Natural Science Foundation of Liaoning Province (20170540288); Special Fund of Dalian city for Distinguished Young Scholars (2017RJ07) and National Key R＆D Program of China (2017YF0200900). Declaration of Interests: The authors declare that they have no competing interests. Ethics Approval Statement: Animal care and experimental procedures were performed in accordance with a protocol approved by the Institutional Animal Care and Ethics Committee of Dalian Medical University (AEE18033). In addition, all experiments in the present study were consistent with the National Institutes of Health guide for the care and use of Laboratory animals.

Expression of Target Genes in Cumulus Cells Derived from Human Oocytes with and without Blastocyst Formation

Objective: The transcriptional profile of cumulus cells (CCs) during oocyte maturation provides information for predicting oocyte developmental competence. Our previous study using a mouse model indicated that there were nine different genes related to oocyte development potential expressed in CCs during oocyte maturation. The purpose of this study was to elucidate whether gene expression levels of CCs during oocyte maturation are associated with oocyte developmental competence. Methods: The human CCs collected from each oocyte were divided into two groups after tracking depending on whether or not they developed to the blastocyst stage: (1) the oocytes were developed to blastocyst stage after fertilization (B+) and (2) the oocytes were not developed to blastocyst stage after fertilization (B−). The expression levels of the nine selected genes (ARRB1, ATP2C1, CDH5, CNTNAP1, LGR4, MKLN1, RHOBTB1, SIX2, and SMC2) were examined. CCs were obtained from 29 women who were undergoing intracytoplasmic sperm injection treatment cycles. Quantitative reverse transcriptase-polymerase chain reaction analysis was performed on cumulus masses collected before insemination. Each sample was run three times. Statistically significant differences in mRNA expression of the target genes in independent samples were evaluated by two-tailed Student's t-test, and P Results: There were significant differences in the mRNA expression levels of ARRB1 (P = 0.016), LGR4 (P = 0.025), and SMC2 (P = 0.013) between the groups B+ and B−. Gene expression of ARRB1, LGR4, and SMC2 in CCs is related to blastocyst development. Conclusions: Analysis of expression of ARRB1, LGR4, and SMC2 genes in CCs as biomarkers may provide predictive information on oocyte developmental competence before insemination and fertilization.

108 Intralesional Collagenase Clostridium Histolyticum Causes Meaningful Improvement in Men with Peyronie’s Disease: The Results of a Multi-institutional Analysis

Peyronie’s disease (PD), defined as the abnormal formation of a fibrous plaque(s) in the tunica albuginea of the penis, is a chronic and distressing condition that affects 3-13% of the adult male population. Collagenase Clostridium histolyticum (CCH) is an injectable agent that enzymatically degrades the abnormal interstitial collagen in PD plaques, and was documented in the IMPRESS trials to reduce penile curvature by an average of 35%. Since these trials, a number of single center studies have been conducted to evaluate the safety and efficacy of CCH in different settings with varying protocols. However, there have not been any published studies analyzing the effect of CCH on patients with PD in a multi-institutional analysis that follows the IMPRESS trials protocol. To evaluate, in a real-world setting, the efficacy/safety outcomes of multiple high-volume centers employing CCH to treat PD. Retrospective data were collected for consecutive patients with PD who underwent treatment with CCH between April 2014 and March 2018 at five institutions. 918 patients were included. Main outcomes of interest include the change in curvature after receiving CCH therapy, and frequency of serious treatment-related adverse events. Other variables of interest included patient age, penile curvature measurements (defined as the largest single plane curvature), curvature direction, plaque location, duration of PD, and number of CCH cycles received. Two-tailed paired Student t-test was used for comparisons of continuous variables. Univariate and multivariate logistic regression analyses were performed to assess predictors of success of CCH therapy in improving curvature, defined as a ≥20% improvement in curvature, and global response scores.

Lower rate of delayed graft function is observed when epidural analgesia for living donor nephrectomy is administered

BackgroundThe beneficial effects of epidural analgesia (EDA) in terms of pain control and postoperative convalescence are widely known and led to a frequent use for patients who underwent living donor kidney nephrectomy. The objective of this study was to determine whether general anesthesia (GA) plus EDA compared to GA only, administered for living donor nephrectomy has effects on postoperative graft function in recipients.MethodsIn this monocentric, retrospective cohort analysis we analyzed the closed files of all consecutive donor- recipient pairs who underwent living donor kidney transplantations from 2008 to 2017. The outcome variable was delayed graft function (DGF), defined as at least one hemodialysis within seven days postoperatively, once hyperacute rejection, vascular or urinary tract complications were ruled out. Statistical analyses of continuous variables were calculated using the two-tail Student’s t test and Fisher exact test for categorical variables with a significance level of p < 0.05, respectively.ResultsThe study enclosed 291 consecutive living donor kidney transplantations. 99 kidney donors received epidural analgesia whereas 192 had no epidural analgesia. The groups showed balanced pretransplantational characteristics and comparable donors´ and recipients’ risk factors. 9 out of all 291 recipients needed renal replacement therapy (RRT) during the first 7 days due to delayed graft function; none of these donors received EDA. The observed rate of DGF in recipients whose kidney donors received epidural analgesia was significantly lower (0% vs. 4.6%; p = 0.031).ConclusionsIn our cohort we observed a significantly lower rate of DGF when epidural analgesia for donor nephrectomy was administered. Due to restrictions of the study design this observation needs further confirmation by prospective studies.

Physiological and anatomical aspects of the reproduction of mice with reduced Syndecan-1 expression

BackgroundSyndecan-1 is a heparan sulfate proteoglycan acting as a co-receptor for cytokines and growth factors mediating developmental, immunological and angiogenic processes. In human, the uteroplacental localization of Syndecan-1 and its reduced expression in pregnancy-associated pathologies, such as the intrauterine growth restriction, suggests an influence of Syndecan-1 in embryo-maternal interactions. The aim of the present study was to identify the effect of a reduced expression of Syndecan-1 on the reproductive phenotype of mice and their progenies.MethodsReproductive characteristics have been investigated using animals with reduced Syndecan-1 and their wildtype controls after normal mating and after vice versa embryo transfers. Female mice were used to measure the estrus cycle length and the weight gain during pregnancy, as well as for histological examination of ovaries. Male mice were examined for the concentration, motility, viability and morphology of spermatozoa. Organs like heart, lung, liver, kidney, spleen, brain and ovaries or testes and epididymis of 6-month-old animals were isolated and weighed. Statistical analyses were performed using two-tailed students t-test with P < .05 and P < .02, chi square test (P < .05) and Fisher’s Exact Test (P < .05). A linear and a non-linear mixed-effects model were generated to analyze the weight gain of pregnant females and of the progenies.ResultsFocusing on the pregnancy outcome, the Syndecan-1 reduced females gave birth to larger litters. However, regarding the survival of the offspring, a higher percentage of pups with less Syndecan-1 died during the first postnatal days. Even though the ovaries and the testes of Syndecan-1 reduced mice showed no histological differences and the ovaries showed a similar number of primary and secondary follicles and corpora lutea, the spermatozoa of Syndecan-1 reduced males showed more tail and midpiece deficiencies. Concerning the postnatal and juvenile development the pups with reduced Syndecan-1 expression remained lighter and smaller regardless whether carried by mothers with reduced Syndecan-1 or wildtype foster mothers. With respect to anatomical differences kidneys of both genders as well as testes and epididymis of male mice with reduced syndecan-1 expression weighed less compared to controls.ConclusionsThese data reveal that the effects of Syndecan-1 reduction are rather genotype- than parental-dependent.

Abstract P1-20-04: Nanoformulation of doxorubicin inside H- ferritin nanocages allows a cardio-safe combined therapy with trastuzumab: De-escalating cardiotoxicity in HER2-positive breast cancer

Abstract Background: HER2+ breast cancer (BC) accounts for 20-25 % of BCs and it is characterized by high aggressiveness.1 Despite the anti-HER2 monoclonal antibody Trastuzumab (TZ) has improved BC clinical outcome, it could induce severe cardiac reversible dysfunction:2 HER2 signaling is also essential for growth and survival of myocardiocytes.3 Therefore, the concurrent use of TZ with other cardiotoxic drugs as doxorubicin (DOX) is discouraged.3 Both neoadjuvant and adjuvant clinical trials have challenged the notion that TZ should be administered with anthracyclines only sequentially,4 despite pre-clinical studies demonstrated the significant enhancement of efficacy by their coadministration.5-7 Nanomedicine answers to this clinical issue with HFn-DOX: a natural protein-based DOX nanoformulation with native tumor targeting capability that displays a self-triggered nuclear delivery of DOX improving antitumor efficacy and reducing both chemoresistance and cardiotoxicity.8 Methods: HER2+ BC bearing mice have been treated 5 times twice a week with placebo, HFn-DOX (1 mg/Kg, i.v.), TZ (5 mg/Kg, i.p.) and with the combination of them. Main end-point were cardiotoxicity and anticancer efficacy. Tumor size was measured by caliper, while antitumor activity and cardiotoxicity were characterized by ICH, immunofluorescence, cytofluorimetry, TEM, mass spectrometry and western blot on resections. Statistical analyses were conducted using two-tailed Student's t-test (P&amp;lt; 0.05) Results: Although single treatments with HFn-DOX or TZ display a good capability to reduce tumor progression, their combination improves antitumor potential, affecting tumor size and angiogenesis. Since the main TZ activity is the induction of the Antibody-Dependent Cell mediated Cytotoxicity, we have assessed the effect of HFn-DOX on Tumor Infiltrating Lymphocytes (TIL), revealing that both TILs enumeration and TIL activity is unaffected by HFn-DOX. On the other hand, HFn-DOX increases the induction of apoptosis, suggesting that the reduction of the tumor size observed in mice treated with the combination of TZ and HFn-DOX is attributable to the coupling of these activity. Mitochondrial morphology has been checked for cardiotoxicity. A pathological increase in mitochondria area coupled with cristae depletion has been evidenced only in mice treated with TZ alone, confirming the overall safety of the HFn-DOX formulation. Interestingly, mice treated with the TZ and HFn-DOX did not display evidences of cardiac suffering. TZ quantification in tumor and heart revealed that the combination with HFn-DOX couples the increased TZ accumulation and penetration in tumor with TZ reduction in heart, resulting in the lack of cardiotoxicity. Conclusions: Our results suggest that a combined therapy with HFn-DOX and TZ allows an enhanced anticancer activity and reduced cardiotoxicity, with potential translational implications on the treatment of HER2+ BC patients. [1] Nat Rev Cancer 2009; 9:463; [2] Nat. Med. 2012; 18(11):1639; [3] Cancer treatments reviews 2009; 35:633; [4] The breast 2014; 23:317; [5] Cancer Res. 1998; 58(13):2825; [6] Lancet Oncol. 2011; 12(3):236; [7] Lancet 2010; 375(9712):377. [8] Oncotarget. 2017; 8:8383. Citation Format: Mazzucchelli S, Andreata F, Bonizzi A, Monieri M, Bellini M, Longhi E, Ottria R, Sorrentino L, Truffi M, Prosperi D, Zerbi P, Corsi F. Nanoformulation of doxorubicin inside H- ferritin nanocages allows a cardio-safe combined therapy with trastuzumab: De-escalating cardiotoxicity in HER2-positive breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P1-20-04.

Agricultural Expansion-Induced Infiltration Rate Change in a West African Tropical Catchment

Land use and land cover in the Dano catchment is characterized by a rapid conversion from seminatural vegetation (fallow) to agriculture (cropland). The study compares both the saturated (Ks) and the unsaturated (Kh) hydraulic conductivities under cropland and fallow in the catchment to gain insights into the effect of the current land use on soil water dynamics. Hydraulic conductivity was measured under forty-two (42) pairs of adjacent cropland-fallow plots using a Hood infiltrometer. Ks, Kh, bulk density, and soil texture were further compared using a paired two-tailed Student’s t-test (p=0.05). The results showed that both Ks and Kh are highly variable irrespective of the land use type (coefficient of variation &gt; 100%). The results also showed that Ks was significantly higher (1.16-fold on average) under fallow compared to cropland. As for Kh, the results showed that, from −2 cm to zero tension heads (h), Kh under cropland and fallow is not significantly different; however, as the supplied tension decreases up to the saturation state, Kh under fallow becomes statistically higher compared to cropland. No significant difference was found between soil textures and bulk density under cropland and fallow meaning that the observed differences of Ks and Kh under cropland and fallow were caused by land use and not preexisting difference in texture. These results suggest an increasing risk of erosion, soil fertility reduction, and flood in the catchment because of agricultural land expansion.

The effect of single pretreatment with salbutamol on recovery of bronchoalveolar lavage fluid in horses with suspected or confirmed severe equine asthma.

BackgroundBronchoalveolar lavage (BAL) is a method for the recovery of respiratory secretion from the lower airways.ObjectivesTo investigate if the administration of a single dose of a bronchodilatator in horses with a suspected or confirmed severe equine asthma could improve recovery of bronchoalveolar lavage fluid (BALF).AnimalsTwenty‐eight horses with severe equine asthma.MethodsHorses were divided into 2 groups: group “treated” was given salbutamol before endoscopic examination and BALF collection, whereas group “not treated” was not given. BAL was performed with BAL‐catheter by instilling 350 mL of sterile saline. Amount of recovered fluid was recorded. Statistical analysis was performed with a two‐tailed Student's t test.ResultsThe average fluid recovery in the horses treated with salbutamol was 52% ± 15% (mean +/− SD), compared with 38% ± 13% for the group of horses not treated with salbutamol (P = 0.013).Conclusions and Clinical ImportanceClinicians should consider administration of salbutamol before performing BAL on horses with asthma.

Development of an indirect ELISA based on whole cell Brucella abortus S99 lysates for detection of IgM anti-Brucella antibodies in human serum.

Brucellosis is the most common zoonotic diseases worldwide. The aim of this study was to develop and evaluate the diagnostic performance of an indirect-ELISA (I-ELISA) method based on whole cell Brucella abortus S99 lysates for detection of IgM anti-Brucella antibodies in a human serum. The study was conducted in two species-rich endemic areas of Iran (Tehran and Lorestan provinces). Serum samples of 102 patients and 150 healthy individuals were tested by the new kit and the commercial Vircell kit for the presence of anti-Brucella IgM antibodies. The disease status was confirmed by Wright agglutination test. The difference in the mean optical densities (OD) recorded by the new and the Vircell kits for patients and healthy individuals were tested using Two-tailed Student t-test. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the new kit were informed using Receiver operating curve analysis. The results were used to guide the choice of cutoff. Agreements in ODs recorded by the new and the Vircell kit was visually inspected using Bland-Altman plot. The new I-ELISA showed excellent diagnostic performance (sensitivity and PPV = 95.7%, specificity and NPV = 97.8%) for the diagnosis of brucellosis. The cut-off area for the antibody index (AI) was determined to be 8-10, where AIs less than 8 and greater than 10 were considered Brucella-negative and -positive, respectively. AIs of 8-10 show equivocal results, requiring re-testing. The Vircell kit showed low (36.8%) sensitivity and perfect (100%) specificity on the same samples. The Bland-Altman plot showed low agreement between both tests in recording the OD values for the same individuals. The new I-ELISA based on whole cell Brucella abortus S99 showed a good performance for the detection of Brucella spp. Lack of agreement between the new and the Vircell kit suggest that the performance of ELISA kits might be dependent on the geographical area under study. Hence, validation of the new and the Vircell kits is recommended prior to their implementation in other geographical areas.

Timing and Outcomes of Abdominal Surgery in Neutropenic Patients

BackgroundSurgery in neutropenic patients is challenging due to both atypical manifestations of common conditions and higher perioperative risk. We sought to describe the outcomes of neutropenic patients undergoing abdominal surgery and to identify factors contributing to morbidity and mortality. MethodsA retrospective chart review was performed for all patients neutropenic in the 24-hours prior to an abdominal operation at our institution between 1998 and 2017. The primary and secondary outcomes were 30-day mortality and morbidity, respectively. The chi-square test and two-tailed Student’s t test were used for univariable comparisons (non-parametric tests used when appropriate). To determine the optimal threshold of absolute neutrophil count (ANC) to discriminate 30-day mortality, we maximized the Youden index (J). ResultsAmongst 237 patients, mortality was 11.8% (28/237) and morbidity 54.5% (130/237). Absolute neutrophil count < 500 cells/μL (50% vs. 20.6%, P < 0.01) and perforated viscus (35.7% vs. 14.8%, P = 0.01) were associated with mortality. Perforated viscus (25.4% vs. 7.5%) was also associated with morbidity. Urgent operations were associated with higher morbidity (63.6% vs 34.7%, P < 0.001) and mortality (16.4% vs 1.4%, P = 0.002) when compared to elective operations. Transfer from an outside hospital (22.3% vs. 11.2%, P = 0.02) and longer median time from admission to operation (2 days (IQR 0–6) vs. 1 day (IQR 0–3), P < 0.01) were associated with morbidity. An ANC threshold of 350 provided the best discrimination for mortality. ConclusionsElective surgery in the appropriately chosen neutropenic patient is relatively safe. For patients with obvious surgical pathology, we advocate for earlier operation and a lower threshold for surgical consultation in an effort expedite the diagnosis and necessary treatment.