Mann-Whitney Rank Sum Test Research Articles

See-and-Treat Loop Electrosurgical Excision Procedure for High-Grade Cervical Cytology: Are We Overtreating?

To report the overtreatment rate for see-and-treat versus 3-step conventional strategy (cervical cytology, colposcopic biopsies, then LEEP) for patients with high-grade squamous intraepithelial lesion (HSIL) cytology. Our second aim was to identify risk factors for overtreatment. We included 178 women with HSIL cytology from our university-based colposcopy clinic who underwent LEEP between 2007 and 2014. Overtreatment was defined as cervical intraepithelial neoplasia (CIN) 1 or less on LEEP specimen. Differences between treatment groups were compared using chi-square test, 2-sample t test, or Mann-Whitney rank-sum test as appropriate. CIN2+ was found in 69 (80%) of women in the see-and-treat group and 69 (75%) of the conventional management group (P = 0.093), with overtreatment in 17 (20%) and 23 (25%, P = 0.403), respectively. Women who underwent see-and-treat (n = 86) were older (mean age, 36 vs 31 years; P = 0.007), and a greater proportion completed childbearing (30% vs 13%, P = 0.024). There were no differences in top hat excision; however, a higher proportion of the see-and-treat group had CIN2+ in endocervical samples (54% vs 27%, P = 0.047). Overtreatment, regardless of management strategy, was associated with age at time of LEEP, where older women were more likely to be overtreated (median age, 37 vs 32 years, respectively; OR, 1.04; 95% CI, 1.01-1.08; P = 0.011). A see-and-treat strategy minimizes risk of loss to follow-up with a similar overtreatment rate compared with conventional management. With CIN2+ in some three-fourths of women with HSIL, a see-and-treat should be favored especially when adherence to follow-up is questionable.

Blood Biomarkers of Chronic Inflammation in Gulf War Illness.

BackgroundMore than twenty years following the end of the 1990–1991 Gulf War it is estimated that approximately 300,000 veterans of this conflict suffer from an unexplained chronic, multi-system disorder known as Gulf War Illness (GWI). The etiology of GWI may be exposure to chemical toxins, but it remains only partially defined, and its case definition is based only on symptoms. Objective criteria for the diagnosis of GWI are urgently needed for diagnosis and therapeutic research.ObjectiveThis study was designed to determine if blood biomarkers could provide objective criteria to assist diagnosis of GWI.DesignA surveillance study of 85 Gulf War Veteran volunteers identified from the Department of Veterans Affairs Minnesota Gulf War registry was performed. All subjects were deployed to the Gulf War. Fifty seven subjects had GWI defined by CDC criteria, and 28 did not have symptomatic criteria for a diagnosis of GWI. Statistical analyses were performed on peripheral blood counts and assays of 61 plasma proteins using the Mann-Whitney rank sum test to compare biomarker distributions and stepwise logistic regression to formulate a diagnostic model.ResultsLymphocyte, monocyte, neutrophil, and platelet counts were higher in GWI subjects. Six serum proteins associated with inflammation were significantly different in GWI subjects. A diagnostic model of three biomarkers—lymphocytes, monocytes, and C reactive protein—had a predicted probability of 90% (CI 76–90%) for diagnosing GWI when the probability of having GWI was above 70%.SignificanceThe results of the current study indicate that inflammation is a component of the pathobiology of GWI. Analysis of the data resulted in a model utilizing three readily measurable biomarkers that appears to significantly augment the symptom-based case definition of GWI. These new observations are highly relevant to the diagnosis of GWI, and to therapeutic trials.

Adrenergic Effect on Cytokine Release After Ex Vivo Healthy Volunteers' Whole Blood LPS Stimulation.

Catecholamines are molecules with immunomodulatory properties in health and disease. Several studies showed the effect of catecholamines when administered to restore hemodynamic stability in septic patients. This study investigates the effect of norepinephrine and dobutamine on whole blood cytokine release after ex vivo lipopolysaccharide (LPS) stimulation. Whole blood collected from healthy individuals was stimulated with LPS, in the presence of norepinephrine or dobutamine at different concentrations, with or without metoprolol, a β1 receptor antagonist. Cytokine measurement was performed in isolated cell culture supernatants with ELISA. Results are expressed as mean ± SEM and compared with Mann-Whitney rank-sum test. Both norepinephrine and dobutamine significantly reduced TNF-α and IL-6 production after ex vivo LPS stimulation of whole blood in a dose-dependent manner, and this effect was partially reversed by the presence of metoprolol. Norepinephrine and dobutamine reduce the LPS-induced production of pro-inflammatory cytokines, thus possibly contributing to altered balance between the inflammatory and anti-inflammatory responses, which are vital for a successful host response to severe disease, shock, and sepsis.

Abstract P1-01-17: Integrative pathology: Analysis of cellular multiplex technology to detect proteomic, genomic and DNA data from fine needle aspiration biopsy specimens

Abstract Background: An integrative system capable of detecting proteomic, genomic and DNA content from cell isolates obtained by fine needle aspiration (FNA) biopsy may offer distinct advantages in diagnosing breast cancer and monitoring response to therapy. Cellular Multiplex™ is such a system. An initial pilot study evaluating this technology established a series of variables that could separate normal from cancerous elements using cells obtained from an FNA performed on excised tumors and reduction mammoplasty specimens. In order for the technology to be clinically relevant, it must perform robustly on intact tumors. The current study was therefore undertaken to validate Cellular Multiplex™ on cells obtained by FNA performed on intact tumor at the time of diagnosis. Methods: Patients undergoing lumpectomy requiring either needle or 125I seed localization were identified. FNA was performed on intact tumor (A samples) at the time of radiographic localization prior to lumpectomy and repeated on the excised tumor (B samples). Cells obtained by FNA were placed in a proprietary fixative then hybridized and stained to detect multiple mRNA and protein targets along with DNA content. Estrogen receptor, progesterone receptor and HER2 were included in the panel of targets and compared to the routine clinical pathology report. Cell morphology was assessed by mean corpuscular volume. Samples were analyzed using an EC800 (Sony Biotechnology, San Jose, CA) and the results from matched A and B samples were compared using the Mann-Whitney Wilcoxson Rank Sum test. The study is designed to enroll 50 patients. Here we report an analysis of the first 9 cases. Results: The cell number obtained from the excised tumors were 3-4 times greater (median to median) than obtained from the intact tumor. There was no statistical difference in the expression of the 2 mRNA targets, 8 protein targets, DNA content and cell morphology between the A and B samples. The parameters derived from Cellular Multiplex matched the standard pathologic features reported on the clinical pathology report in 8 of 9 cases. In the one discrepant case, Cellular Multiplexing detected ER positive cells in a case where standard pathologic evaluation with immunohistochemistry reported the tumor to be estrogen receptor negative. Conclusions: This interim analysis demonstrates that the Cellular Multiplex technology is working well using cells obtained by FNA performed on intact tumors with readouts matching those obtained from excised specimens. If confirmed in the remaining patients, these data suggest that this technology will be applicable for the evaluation of intact tumors thereby making it relevant for multiple clinical indications including diagnosis and monitoring response to neoadjuvant therapy. Citation Format: Mittendorf EA, Dogan B, Morgan R, Chargin A, Wu Y, Cornett-Risher S, Shults K. Integrative pathology: Analysis of cellular multiplex technology to detect proteomic, genomic and DNA data from fine needle aspiration biopsy specimens. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P1-01-17.

Abstract A33: Dysfunction of subtelomeric methylation and telomere length in different grades of gallbladder cancer

Abstract Background: Gallbladder cancer (GBC), although a rare cancer, has a very high incidence in certain parts of the world including North Central India. A recent report from our lab has shown a very high incidence in the Gwalior-Chambal region of North Central India, ranking fourth among all cancers and first among gastrointestinal tract cancers. Due to its highly aggressive nature and inadequate therapeutic alternatives, its mortality is very high while prognosis is poor. Gallstones, which are an important risk factor, are present in most of the tumor cases. Telomeres are known to protect the organism against chromosomal instability and its dysfunction has been reported to be an early and common event in pre-cancerous lesions. Subtelomeric repeats, which are located adjacent to telomeres, include sequences, such as D4Z4 and DNF92. Subtelomeric methylation has been recently reported to negatively regulate telomere length in various diseases, including cancer. However, there is no report on the influence of subtelomeric methylation in telomere dysfunction in GBC and whether this is affected by the presence of gall stones. Materials and Methods: Monochrome multiplex quantitative PCR was used to determine telomere length in 37 pairs of adjacent non tumor and adenocarcinoma tissues and 37 gall stone tissues. Methylation status of 2 subtelomeric sequences, D4Z4 and DNF92 was assessed by bisulfite sequencing in 24 samples of different tissue grades. Comparison was made between normal, gallstone and different grades of cancer tissues with regard to their telomere length and methylation status. To calculate statistical significance of the observed differences in telomere length and methylation status, Mann-Whitney Rank Sum test and Chi square test was applied, respectively. Linear regression analysis was applied to test the correlation between telomere length and subtelomeric methylation. Results: Telomere length was observed to be shortened significantly in early grade GBC (p < 0.001) and not in late grade. Also, D4Z4 and DNF92 sequences were found to be hyper- and hypo-methylated, respectively, in GBC (D4Z4, p = 0.01; DNF92, p = 0.01), with their levels significantly differing only in early grades of GBC (D4Z4, p = 4x10-6; DNF92, p = 0.01). Surprisingly, no correlation could be found between subtelomeric methylation levels and telomere length. Interestingly, telomere length and subtelomeric methylation differed significantly in tumor tissues without gallstones and not in tumor tissues with gallstones (telomere length, p=0.02; D4Z4, p = 0.005; DNF92, p = 0.009). Conclusion: This study, thus, suggests that telomere dysfunction and changes in methylation levels occurs earlier in the progression of GBC and presence of gall stones may have no effect on telomere length as well as methylation levels. Citation Format: Satish Poojary, Gunja Mishra, Sanjiv Gupta, Braj Raj Shrivastav, Pramod Kumar Tiwari. Dysfunction of subtelomeric methylation and telomere length in different grades of gallbladder cancer. [abstract]. In: Proceedings of the Fourth AACR International Conference on Frontiers in Basic Cancer Research; 2015 Oct 23-26; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2016;76(3 Suppl):Abstract nr A33.

Clinical Outcome of (131)I Therapy in Differentiated Thyroid Cancer with Preablative Thyroglobulin below 10 ng/ml.

To investigate the clinical outcome of (131)I therapy in differentiated thyroid cancer (DTC) with preablative stimulated thyroglobulin (ps-Tg) below 10 ng/ml and the value of ps-Tg for predicting clinical outcome. Totally 167 DTC patients with ps-Tg below 10 ng/ml were included and divided into three groups [excellent response (ER, n=131), acceptable response (AR, n=34), and incomplete response (IR, n=2)] according to the response to initial (131)I therapy. One-way analysis of variance, χ(2) test, and Kruskal-Wallis test were used to evaluate the statistical differences of clinicopathological features, recurrence risk, and ps-Tg value among the three groups. The ps-Tg value between ER group and non-ER group were compared by Mann-Whitney rank-sum test. The ROC curve and optimal cut-off point were analyzed to evaluate the clinical value of ps-Tg for predicting disease-free status. There was significant difference in ps-Tg level (H=14.305, P=0.001) among three groups but not in age (F=0.831, P=0.755), sex (χ(2)=0.178, P=0.915), cervical lymph node metastases (χ(2)=1.475, P=0.478), TNM stage (H=1.063, P=0.588), and recurrence risk (H=2.947, P=0.229). The median level of ps-Tg in ER group was 2.20 ng/ml (1.10, 4.40), which was significantly lower than 4.40 ng/ml (2.70, 5.90) of non-ER groups (U=1424.50, P=0.000). Area under the ROC curve was 0.717. The cut-off value of ps-Tg was 2.35 ng/ml,with a sensitivity of 83.33%, specificity of 53.43%, and negative predictive value of 92.11%. The serum ps-Tg value is a sensitive marker for predicting the disease-free status in DTC patients with ps-Tg below 10 ng/ml.

The Effect of Eye Drop Technique Education in Patients With Glaucoma

ABSTRACTEducation about how to administer eye drops may improve a patient’s ability to instill his or her eye drops correctly. Our objectives were to (a) document the methods providers use to educate glaucoma patients about eye drop technique; (b) determine whether eye drop technique education varies by provider and patient characteristics; and (c) evaluate whether education predicts improved patient technique. We conducted an 8-month longitudinal study of 279 glaucoma patients and 15 providers in which we recorded on videotape the content of glaucoma office visits at two time points (baseline and 4- to 6-week follow-up) and videotaped patient eye drop technique at three time points (baseline, 4- to 6-week follow-up, and 8-month follow-up). Mann–Whitney rank sum tests were used to determine whether education was associated with improved patient eye drop technique over time. Ninety-four patients (34%) received technique education at either visit; 31% received verbal education and 10% received a technique demonstration. Only 24 patients (47%) who were new to eye drops received technique education at the baseline visit. Patients who were new to drops at baseline (p = .008) and patients who asked a question about drops (p < .001) were more likely to receive technique education. Education was not associated with improved technique. Eye drop technique education occurs infrequently during glaucoma office visits. Future studies should compare the effectiveness of different educational methods, such as patient demonstration versus provider verbal instruction, to determine which method is best at improving patient eye drop technique.

Particulate-bound mercury (PBM) in stems and leaves of several crops (white cabbage, Peking cabbage, and chili) at a farmland site in Taiwan

ABSTRACTThis study analyzes the particulate-bound mercury (PBM) contents of two parts (leaf and stem) of different crops (white cabbage, Peking cabbage, and chili) at a sampling site in the coastal zone in the Taichung area from October 7 to 15, 2014. A direct mercury analyzer (DMA-80) was used to measure mercury content. The Mann-Whitney rank sum test was utilized to determine the mean differences between the PBM in two parts of various crops (white cabbage leaf, white cabbage stem, Peking cabbage leaf, Peking cabbage stem, and chili) at the Long-Jing sampling site. The results indicated the following: 1) The average mercury content was lowest in the Peking cabbage stem. The ratio of the mercury content in the stem to that in the leaf of the white cabbage was 5 and that of Peking cabbage was 3.5.2) No significant difference was observed between median mercury content in white cabbage leaf and Peking cabbage leaf, white cabbage stem and Peking cabbage stem, and Peking cabbage leaf and chili. 3) The average PBM contents were both the lowest (Huabei Plain) and the highest in Hg in China. 4) The ratio of average PBM content of white cabbage leaf in this study was as high as 981.

G-111 DNA prime/subunit boost using SIVE660 based rhFLSC yields 75% efficacy against cross clade SIVmac251 intrarectal challenge

An effective vaccine for HIV still remains elusive. Our previous studies showed that a protein-based FLSC (a genetic fusion of gp120 and CD4) protected macaques against repeat rectal challenges with SHIV162P3 but efficacy was low. We hypothesized that DNA would provide a superior prime to a subunit boost than subunit alone and that the adjuvants IL-12 and/or LTA1 would improve priming. FLSC delivery regimens were evaluated in a challenge model using SIVsmE543 antigens and a cross clade challenge with SIVmac251. Eight macaques/group were immunized with DNA expressing a gag/pol fusion and FLSC with/without LTA1, IL-12 or LTA1+IL-12 by electroporation on weeks 0, 4 and 8. Booster immunizations of 300 μg of FLSCsmCG7V/alum were given at week 42. Two weeks later, animals were weekly challenged rectally 10 times with SIVmac251. Immune measures were compared using T tests (parametric) or Mann-Whitney Rank Sum Tests (non-parametric). Infection rates were compared using Log Rank tests. The DNA/IL-12 prime regimen provided 75% efficacy compared to the same regimen without IL-12. Efficacy did not correlate with antibody binding or neutralizing titers to Tier 1 SIVmac251 whereas ADCC titers and antibody binding ratios of FcgR3/FcgR1 correlated with protection, provided T cell responses were low. The DNA/LTA1/IL-12 regimen generated substantially superior CMI responses, but showed no protection. These results suggest that vaccines generating ADCC capable antibodies with high FcgR3/FcgR1 ratios that evoke minimal T cell responses may be the most protective.

Effects of dexmedetomidine on perioperative monitoring parameters and recovery in patients undergoing laparoscopic cholecystectomy.

Background:Dexmedetomidine, an α2 agonist, when used as an adjuvant in general anesthesia attenuates stress response to various noxious stimuli, maintains perioperative hemodynamic stability and provides sedation without adversely affecting recovery in postoperative period.Materials and Methods:Sixty patients were randomly divided into two groups of 30 each. In Group A, dexmedetomidine was given intravenously as loading dose of 1 μg/kg over 10 min, and normal saline was given in Group B patients. After induction with propofol, in Group A, dexmedetomidine was given as infusion at a dose of 0.2–0.8 μg/kg/h. Sevoflurane was used as inhalation agent in both groups. Perioperative monitoring parameters were recorded. Postoperative sedation and recovery were assessed.Statistical Analysis Used:Demographic data were analyzed using Pearson's Chi-square test. Changes in the heart rate (HR), systolic blood pressure (BP) and diastolic BP were analyzed using unpaired t-test and Mann–Whitney rank sum test was used to calculate “P” value wherever (Shapiro–Wilk)/normality test gave ambiguous results.Results:Dexmedetomidine significantly attenuates stress response at intubation with lesser increase in HR (86.00 ± 5.16 vs. 102.97 ± 7.07/min.), mean BP (95.78 ± 5.35 vs. 110.18 ± 5.35) as compared to the control group (P < 0.05). After pneumoperitoneum, HR was 85.07 ± 6.23 versus 107.10 ± 4.98, mean BP was 98.98 ± 10.16 versus 118.54 ± 6.27 (P < 0.05). Thus maintains intraoperative hemodynamic stability. Postoperatively, the test group showed no statistically significant difference in the extubation time (7.00 ± 0.58 vs. 6.74 ± 0.73) and response to oral commands (8.78 ± 0.72 vs. 8.66 ± 0.73) (P > 0.05).Conclusion:Dexmedetomidine attenuates various stress responses during surgery and maintains the hemodynamic stability when used as an adjuvant in general anesthesia and dexmedetomidine does not delay recovery.

Presence of Fleischer ring and prominent corneal nerves in keratoconus relatives and normal controls.

To examine the occurrence of commonly known clinical signs of keratoconus (KC), i.e. Fleischer ring, prominent corneal nerves and thinning, among unaffected family members of KC patients and healthy control individuals. Data of both eyes of 117 relatives of KC patients having no manifest disease based on videokeratography indices (KC relatives), and 142 controls were used for Pearson correlation and t-test statistics. Correlation of Fleischer ring, prominent corneal nerves and central pachymetry data were tested with each other and with videokeratography indices (KSI, KISA, 3 and 6 mm Fourier asymmetry, and I-S). A moderate correlation was found between Fleischer ring and all examined topographical indices. Most important correlation was present with 6 mm Fourier asymmetry, and corneal pachymetry (r=0.272, P<0.001; r=-0.234, P=0.027, respectively). Similar correlations were found with prominent corneal nerves (r=0.234, P<0.001 for 6 mm Fourier asymmetry and r=-0.235, P=0.0265 for pachymetry). KC family members who exhibited Fleischer ring or prominent nerves had thinner and more asymmetric corneas than those without Fleischer ring or prominent corneal nerves (P<0.05 for pachymetry and topographic indices with t-test and Mann-Whitney rank sum test). Though rarely, Fleischer ring and prominent corneal nerves occurred among normal controls, indicating the existence of forme fruste cases in the normal population. Control subjects, who had corneal Fleischer ring or prominent nerves had corneas more similar to KC than other controls (t-test: increased KSI and KISA, P=0.048 and 0.012, respectively). In KC family members and healthy individuals, Fleischer ring and prominent corneal nerves are associated with features of KC and may suggest a possibility of forme fruste KC. Searching for the possible presence of Fleischer ring or prominent nerves on the cornea may help in the decision whether or not to diagnose subclinical KC in a borderline case.

Employability of Nursing Care Graduates

Abstract Starting points: In Slovenia, the higher education institution for nursing started exploring employability opportunities in nursing care in connection with the achievement of competencies from students’ and employers’ point of view. This article highlights the importance of monitoring nursing graduates’ employability. Its aim is to examine the employability of nursing care graduates based on the self-evaluation of competences obtained during the last study year and to establish a link between the self-evaluation of competences and students’ academic performance. Methodology: A questionnaire was distributed to full and part time nursing care students attending the last study year at five different healthcare/health sciences faculties in Slovenia and to employers (healthcare institutions) where the majority of nursing care graduates finds employment. We examined the level of competence achieved by nursing students and the level of competences required by employers. The sample included a total of 485 students. 194 surveys were returned, which represent a 40 percent response. We used Kolmogorov-Smirnov test for each individual joined competence. Further, we compared employability skills of students and employers with Mann-Whitney and Wilcox rank-sum test. For correlation between two variables we used Spearman correlation analysis. Results: The Mann-Whitney and Wilkson Rank test show that employers generally assess competences with a higher average grade in comparison to students and these differences are statistically significant. By applying the Spearman correlation analysis, we established that a statistically significant weak correlation may be observed between the “average grade” and “competences” variables. Discussion and conclusion: Our findings show that a continuous monitoring of general and subject-specific competences gained by students, along with a periodic verification of competences demanded by employers, is necessary. It is very important to monitor the requirements of the labour market in terms of ongoing communication with employers who can best estimate special knowledge needs.

Solid Microneedles for Transdermal Delivery of Amantadine Hydrochloride and Pramipexole Dihydrochloride.

The aim of this project was to study the influence of microneedles on transdermal delivery of amantadine hydrochloride and pramipexole dihydrochloride across porcine ear skin in vitro. Microchannel visualization studies were carried out and characterization of the microchannel depth was performed using confocal laser scanning microscopy (CLSM) to demonstrate microchannel formation following microneedle roller application. We also report, for the first time, the use of TA.XT Plus Texture Analyzer to characterize burst force in pig skin for transdermal drug delivery experiments. This is the force required to rupture pig skin. The mean passive flux of amantadine hydrochloride, determined using a developed LC–MS/MS technique, was 22.38 ± 4.73 µg/cm2/h, while the mean flux following the use of a stainless steel microneedle roller was 49.04 ± 19.77 µg/cm2/h. The mean passive flux of pramipexole dihydrochloride was 134.83 ± 13.66 µg/cm2/h, while the flux following the use of a stainless steel microneedle roller was 134.04 ± 0.98 µg/cm2/h. For both drugs, the difference in flux values following the use of solid stainless steel microneedle roller was not statistically significantly (p > 0.05). Statistical analysis was carried out using the Mann–Whitney Rank sum test.

Use of a perineural coiled catheter at the sciatic nerve in dogs after tibial plateau levelling osteotomy - preliminary observations.

The analgesic effects of peripheral nerve blocks can be prolonged with the placement of perineural catheters allowing repeated injections of local anaesthetics in humans. The objectives of this study were to evaluate the clinical suitability of a perineural coiled catheter (PCC) at the sciatic nerve and to evaluate pain during the early post‐operative period in dogs after tibial plateau levelling osteotomy. Pre‐operatively, a combined block of the sciatic and the femoral nerves was performed under sonographic guidance (ropivacaine 0.5%; 0.3 mL kg−1 per nerve). Thereafter, a PCC was placed near the sciatic nerve. Carprofen (4 mg kg−1 intravenously) was administered at the end of anaesthesia. After surgery, all dogs were randomly assigned to receive four injections of ropivacaine (group R; 0.25%, 0.3 mL kg−1) or NaCl 0.9% (group C; 0.3 mL kg−1) every 6 h through the PCC. Pain was assessed by use of a visual analogue scale (VAS) and a multi‐dimensional pain score (4Avet) before surgery (T‐1), for 390 min (T0, T30, T60, T120, T180, T240, T300, T360 and T390) as well as 1 day after surgery (Day 1). Methadone (0.1 mg kg−1) was administered each time the VAS was ≥40 mm or the 4Avet was ≥5. At T390 dogs received buprenorphine (0.02 mg kg−1). Data were compared using Mann–Whitney rank sum tests and repeated measures analysis of variance. Regardless of group allocation, 55% of dogs required methadone. VAS was significantly lower at T390 (P = 0.003), and at Day 1 (P = 0.002) and so was 4Avet at Day 1 (P = 0.012) in group R than in group C. Bleeding occurred in one dog at PCC placement and PCC dislodged six times of 47 PCCs placed. Minor complications occurred with PCC but allowed four repeated administrations of ropivacaine or saline over 24 h in 91.5% of the cases.

Thyroid Hormone and Thyrotropin Concentrations and Responses to Thyrotropin Releasing Hormone in Ageing Horses

Reasons for performing studyThyroid hormones (THs) decrease with age in healthy dogs and cats, although they tend to remain within established reference ranges. Thyrotropin (TSH) is increased in elderly people, with or without mild alterations in THs.ObjectivesTo test the hypothesis that geriatric horses will have lower THs and/or higher TSH compared with younger horses.Study designCross‐sectional study.MethodsResting THs and TSH, and responses to thyrotropin releasing hormone (TRH) were compared between young and old horses. Data from 71 normal, healthy horses that had participated in prior research projects were examined, and found to contain 42 horses 3–10 years of age, 16 horses &gt;15 years and 10 horses &gt;20 years. All samples had been assayed in the same previously validated radioimmunoassays. Statistical analysis was performed with commercial software.ResultsAlthough lower, THs were not significantly different between young and old horses when compared with the Mann–Whitney rank sum test. However, TSH concentration was significantly higher in horses aged &gt;20 (median 0.52 ng/ml) or &gt;15 years (0.44 ng/ml) compared to younger horses (0.33 ng/ml). TRH stimulation tests were performed in 19 of the younger horses, 7 of the horses &gt;20, and 11 of the horses &gt;15. Two‐way repeated measures ANOVA on ranks revealed no significant differences in TH responses to TRH. The TSH response to TRH appeared to be slightly greater in the older horses, but did not quite reach significance (P = 0.06).ConclusionSimilar to reports in elderly humans, aged horses have higher serum TSH concentrations than younger horses.Ethical animal research: All horses in this study were part of a NCSU IACUC approved study. Owners gave informed consent for their horses' inclusion in the study. Sources of funding: College of Veterinary Medicine North Carolina State University, USA Equestrian. Competing interests: None declared.

Abstract 379: Circulating tumor cells (CTCs) in clinically localized prostate cancer (PCa): searching a prognostic tool

Abstract Background If PCa screening has reduced advanced disease and disease-specific mortality, the tradeoff is an over-diagnosis of cases that would not have caused clinical consequences during a man's lifetime if left untreated. Conversely, a vast numbers of men, which harbor occult extra-prostatic extension, develop recurrence after surgery. Despite it looks like an addressable question by quantifying Circulating Tumor Cells (CTCs), unlike breast cancer, in which prognostic and predictive impact of tumor cells in peripheral blood or bone marrow (Disseminated Tumor Cells, DTCs) was largely provided, their role in localized PCa is far from clear. Methods From July 2011 to August 2013, we quantify the tumor burden at diagnosis in peripheral blood of 153 PCa patients stage T2a-T3b, enrolled at three clinical sites (Roma, Orbassano and Padova). All the patients were candidate to undergo radical prostatectomy because of positive biopsy for cancer. Two labs (Orbassano and Padova) performed the CellSearch assay. In patients enrolled in Roma (Janus trial, a phase II study for the use of zoledronic acid as neoadjuvant treatment of invasive PCa), we evaluated in parallel the DTCs. Results We used data obtained in patients enrolled in Orbassano (n = 50) and Roma (n = 15) as training set, meanwhile we used patients of Padova (n = 88) as validation set. Consistently with previous reports (Kraan, Sleijfer et al. 2011) reporting low inter-test and inter-lab variability, the baseline CTC count in the two sets did not significantly differ (Mann-Whitney Rank Sum Test, p = 0.170), so that they were further analyzed altogether. Overall, we found that 74 out of 153 PC patients (48,4%) had at least 1 CTC per 7.5 ml of peripheral blood; 45 PC (29,4%) patients had &amp;gt; 2 cells and 21 PC patients (13,7%) had &amp;gt; 3 cells. Interestingly, 8 out of 153 PC patients (5,2%) had &amp;gt; 5 cells at the first blood draw, before surgery. When we synchronously detected CTCs and DTCs, we found consistent results in 64% of patients (Wilcoxon signed Rank Test, p = 0.339). Conclusions Even when the PCa is pathologically organ confined at surgery, it was reported 5% to 20% of these patients harbor foci of micro-metastatic disease that will later manifest itself as recurrent disease. Here we show CTCs by CellSearch assay in close to 50% of biopsy-confirmed PCa patients, obtaining evidence of a systemic disease. Prospective studies will be required to investigate whether CTC-status is associated with worst outcome in the M0 setting of PCa, as already provided in early breast cancer patients (Lucci, Hall et al. 2012). Citation Format: Elisabetta Rossi, Antonella Facchinetti, Vittorio Aneloni, Emanuel Zilio, Massimo Dal Bianco, Alice Zoccoli, Daniele Santini, Diletta Garrou, Francesco Porpiglia, Rita Zamarchi. Circulating tumor cells (CTCs) in clinically localized prostate cancer (PCa): searching a prognostic tool. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 379. doi:10.1158/1538-7445.AM2015-379

Radiation dose during CT-guided percutaneous cryoablation of renal tumors: Effect of a dose reduction protocol

PurposeTo estimate and compare the radiation dose using a standard protocol and that of a dose reduction protocol in patients undergoing CT-guided percutaneous cryoablation of renal tumors. Materials and methodsAn IRB-approved, HIPAA-compliant retrospective study of 97 CT-guided cryoablation procedures to treat a solitary renal tumor in each of 97 patients (64M, 33F; range 31–84 yrs) was performed. Fifty patients were treated using a standard dose protocol (kVp=120, mean mAs=180, monitoring scans every 3min during freezes), and an additional 47 patients were treated using a dose reduction protocol (kVp=100, mean mAs=100, monitoring scans less frequently than every 3min during freezes). Multiple Wilcoxon Mann-Whitney (rank-sum) tests were used to compare dose-length product (DLP) between the two groups. Fisher’s exact test was used to compare technique effectiveness at 12 months post ablation between the two groups. ResultsMedian DLP for the standard protocol group was 4833.5 mGy*cm (range, 1667–8267 mGy*cm); median DLP for the dose reduction group was 2648 mGy*cm (range, 850–7169 mGy*cm), significantly less than that of the standard protocol group (p<0.01). The technique effectiveness for the dose reduction group was not significantly different from that of the standard protocol group at 12 month follow up (p=0.434). ConclusionThe radiation dose during percutaneous CT-guided cryoablation of renal tumors was substantial in both the standard and the dose reduction groups; however, it was significantly lower with the protocol change that reduced dose parameters and decreased the number of CT scans.

The flavonoid beverage Haelan 951 induces growth arrest and apoptosis in pancreatic carcinoma cell lines in vitro

BackgroundA major challenge in pancreatic cancer treatment is the resistance of human pancreatic cancer cells to apoptosis. Soy isoflavones and calpain inhibition have been suggested to exert inhibitory effects on cancer development and progression. We investigated the effects of the isoflavone containing beverage Haelan 951 and the calpain inhibitor PD150606 on the viability, growth and apoptosis of the human pancreatic cancer cell lines CAPAN-1 and BxPC-3, on the rat pancreatic cancer cell line AR42J, and on human fibroblasts as the control cell line.MethodsCellular viability and proliferation were determined using the LDH cytotoxicity and WST-1 assay, respectively. Apoptosis was detected by flow cytometric analyses of Annexin V-FITC labeled-cells, TUNEL assay and caspase activation. Student’s t test or Mann–Whitney Rank Sum test were used to compare the data.ResultsHaelan concentrations lower than 8 % showed no cytotoxic effects, whereas higher concentrations led to necrosis. Eight percent Haelan induced significant growth inhibition of CAPAN-1 and BxPC-3 cell lines by 30 % and 35 %, respectively, compared with the control. The proliferation rate of AR42J cells decreased by 50 %, whereas the fibroblasts remained unaffected. An 1.1-fold increase in apoptosis was found in CAPAN-1 cells, whereas the number of apoptotic BxPC-3 cells was elevated 2-fold. The number of apoptotic AR42J cells and fibroblasts was elevated 1.5-fold, each. Inhibition of calpain activity amplified the Haelan-induced growth inhibition of CAPAN-1 and BxPC-3 cells, but failed to amplify the growth inhibition of Haelan-treated AR42J cells. In fibroblasts, calpain inhibition induced Haelan-independent growth inhibition. Calpain inhibition also amplified the Haelan-induced apoptotic activity in all cancer cell lines, but exerted no further effect in fibroblasts.ConclusionsThe proliferation-inhibiting and apoptosis-inducing effects of Haelan are highly dependent on cell type and concentration administered. The results show for the first time that Haelan may be a promising candidate in the treatment of human pancreatic cancer, and its anticancer activity may be potentiated when administered with calpain inhibitors.

A validation study of a modified Bouchard activity record that extends the concept of ‘uptime’ to Rett syndrome

The aim of this study was to investigate the validity of using a Bouchard activity record (BAR) in individuals with Rett syndrome to measure physical activity, as compared with pragmatic criterion standard measures of walking status and step counts recorded using the StepWatch activity monitor (SAM). During the waking hours of 1 day, 43 females (mean age 21y, SD 9y) wore a SAM whilst a proxy completed a modified BAR. Responses to the BAR were compared among participants, who were grouped according to walking status, using the Mann-Whitney two-sample rank-sum test. Relationships were sought between BAR responses and step counts using linear regression. According to the proxy-reported BAR responses, those who needed assistance with walking spent more time sitting (median [interquartile range] 9h 15min [8h 8min-10h 30min] vs 6h 15min [4h 15min-8h 30min]; p<0.001) and less time standing (1h [38min-1h 30min] vs 2h 15min [45min-3h 45min]; p=0.04) than those who could walk independently. In those who could walk independently, time classified as 'uptime' (standing and walking) using the BAR was associated with increased step count (r(2) =0.58; p<0.001). These data support the validity of proxy-reported BAR responses. In those who could walk independently, uptime, classified using the BAR, could be used to estimate daily step count. This tool offers an inexpensive method for clinicians to gain insights into physical activity levels in individuals with Rett syndrome.

Preliminary field study of soil TKN in a wastewater land application system

Abstract An accurate nitrogen mass balance is necessary when designing a municipal wastewater land application system in order to reduce the potential for nitrogen contamination of groundwater. Limited information on soil total Kjehldahl nitrogen (TKN) is available to estimate the TKN variance in the soil that can be used for calculating the nitrogen mass balance. This study investigated the soil TKN temporal variance at two different depth ranges within three different soil types irrigated with municipal wastewater effluent in a short-term at a long-term running site under field conditions. Three plots were chosen with each plot growing different plants and with different soil types in Lubbock, Texas, USA. Kruskal–Wallis one way analysis of variance on ranks and Mann–Whitney rank sum test were employed to compare the differences of medians of many groups of data and of two groups of data at p