Washout Groups Research Articles

SP1.2.12A meta-analysis of the impact of rectal washout on local recurrence following anterior resection for rectal cancers

Abstract Background Standardisation of surgical technique in form of total mesorectal excision (TME) and the use of preoperative radiotherapy have led to improved oncological outcomes in rectal cancer. However, the effectiveness of rectal washout in reducing local recurrence (LR) following anterior resection remains debatable. The aim of this meta-analysis was to evaluate the effectiveness of rectal washout in reducing incidence of LR after anterior resection for rectal cancer. Methods A literature search of electronic databases including PubMed, Embase, Scopus and Cochrane was performed for studies that compared rectal washout to no washout after anterior resection using TME for rectal cancers. The review included all articles reporting oncological outcome of local recurrence. Meta-analysis was carried out using random effect model. Results A total of 5 studies involving 5315 patients were included in meta-analysis. Median follow-up was 60 (range 33–60) months. Overall local recurrence rate was 6.6 % with no significant difference in LR rate between the rectal washout and no washout groups (5.23% vs. 9 %)(P = 0.93; RR 0.97; 95% CI 0.52- 1.83).Similarly in subgroup analysis including prospective studies only no significant difference in LR was seen after washout (P = 0.06) or in group of patients treated with a curative intent(P = 0.50). LR was significantly lower in patients who had a rectal washout with normal saline (P < 0.00001; RR 0.59; 95% CI 0.47-0.74). Conclusion This meta-analysis shows that there is no benefit of rectal washout in reducing incidence of local recurrence after anterior resection for rectal cancers.

Short-term testosterone use in female mice does not impair fertilizability of eggs: implications for the fertility care of transgender males.

Does testosterone use in females affect reproductive potential, particularly with regard to the production of fertilizable gametes? Testosterone (T) injections given to post-pubertal female mice caused virilization and although the ovaries were smaller than controls they were still responsive and produced fertilizable eggs when superovulated. Studies to examine the effects of testosterone on reproductive potential in transgender males are lacking. Recently, a model was developed that simulates many aspects of testosterone use in transgender males in order to look at reproductive effects of testosterone in female mice. This study found masculinizing effects on the mice but did not find significant deficits on the number of ovarian follicles; however, effects of testosterone use on ovarian stimulation and fertilizability of oocytes were not investigated. A total of 66, 6-week-old Hsd:NSA (CF-1) female mice and six Hsd:ICR (CD-1) male mice were used for this study. Mice were injected s.c. with 400 µg T or sesame oil once a week for 6 weeks and were either killed 1 week after the sixth injection (active exposure group), or 6-7 weeks after the final T injection (washout group). Both active exposure and washout groups were further subdivided into three groups: unstimulated, equine CG (eCG)-stimulated or eCG/hCG-stimulated. eCG-stimulated mice were killed 44-48 h after eCG injection. eCG/hCG-stimulated mice were injected with eCG, followed 48 h later with hCG. Mice were killed ∼13-18 h after the hCG injection. Data collected included daily vaginal cytology, terminal testosterone levels, ovary weights and histology, number of oocytes/eggs collected in each group, and cleavage to the two-cell stage following IVF. Testosterone-treated mice had testosterone levels elevated to the level of male mice and ceased cycling. Ovaries were significantly smaller in testosterone-treated mice, but they contained normal cohorts of follicles and responded to gonadotrophin stimulation by ovulating similar numbers of eggs as controls, that fertilized and cleaved in vitro. Mice were treated for only 6 weeks, whereas many transgender men use testosterone for many years before considering biological children, and developmental competence was not assessed. Importantly, a mouse system may not perfectly simulate human reproductive physiology. The current standard of care for transgender men who desire biological children is to cease testosterone therapy prior to ovarian stimulation, but the necessity for stopping testosterone is not known. Our model demonstrates that it is possible for testosterone-suppressed ovaries to respond to gonadotrophic stimulation by producing and ovulating fertilizable eggs, thereby obviating the need for testosterone cessation prior to ovarian stimulation. In time, these results may provide insights for future clinical trials of fertility treatment options for transgender men. This study was funded by the Reproductive Endocrinology and Infertility fellowship program through UConn Health Graduate Medical Education (to C.B.B.). The authors have no competing interests. N/A.

Effects of gestational and breastfeeding caffeine exposure in adenosine A1 agonist-induced antinociception of infant rats.

Caffeine is extensively consumed as a psychostimulant drug, acting on A1 and A2A adenosine receptors blockade. Chronic exposure to caffeine during gestation and breast-feeding may be involved in infant rat's behavioral and biochemical alterations. Our goal was to evaluate the effect of chronic caffeine exposure during gestation and breast-feeding in the functionality of adenosine A1 receptors in infant rats at P14. NTPDase and 5'-nucleotidase activities were also evaluated. Mating of adult female Wistar rats was confirmed by presence of sperm in vaginal smears. Rats were divided into three groups on the first day of pregnancy: (1) control: tap water, (2) caffeine: 0.3g/L until P14, and (3) washout caffeine: caffeine was changed to tap water at P7. Evaluation of nociceptive response was performed at P14 using hot plate (HP) and tail-flick latency (TFL) tests. A1 receptor involvement was assessed using caffeine agonist (CPA) and antagonist (DPCPX). Enzymatic activities assays were conducted in the spinal cord. Gestational and breastfeeding exposure to caffeine (caffeine and washout groups) did not induce significant alterations in thermal nociceptive thresholds (HP and TF tests). Both caffeine groups did not show analgesic response induced by CPA when compared to the control group at P14, indicating chronic exposure to caffeine in the aforementioned periods inhibits the antinociceptive effects of the systemic A1 receptor agonist administration. No effect was observed upon ectonucleotidase activities. Our results demonstrate that chronic caffeine exposure in gestational and breastfeeding alters A1-mediated analgesic response in rats.

α4-integrin receptor desaturation and disease activity return after natalizumab cessation.

Objective:To describe the time course of α4-integrin receptor desaturation and disease activity return in patients with relapsing-remitting MS who discontinued natalizumab and to investigate baseline and on-study predictors for the recurrence of disease activity.Methods:In the course of TOFINGO, a 32-week, patient- and rater-blinded multicenter, parallel-group study, we performed MRI, counted relapses, and measured α4-integrin receptor occupancy (RO) at baseline and 8, 12, 16, 20, and 24 weeks. The relationship between RO and total number of new T1 gadolinium-enhancing (Gd+) lesions was modeled using Poisson linear regression.Results:Patients (N = 142) were randomized (1:1:1) to 8-, 12-, or 16-week washout (WO) groups. At randomization, the median RO in the 8-, 12-, and 16-week WO groups was 94.5%, 92.4%, and 90.9%, which declined to 79.8%, 30.7%, and 8.7% after 8, 12, and 16 weeks of WO, respectively. The percentage of patients with new T1 Gd+ lesions increased with longer WO period before commencing fingolimod: 2.1% (8 weeks), 9.1% (12 weeks), and 50.0% (16 weeks). Overall, 71% of patients with first relapse between weeks 6 and 18 had RO values below the time-matched population median. Higher T2 lesion volume (LV) at baseline predicted a higher number of new T1 Gd+ lesions.Conclusions:A faster decline in natalizumab RO, longer WO period, and higher T2 LV at baseline were associated with an increased risk for return of inflammatory disease activity. These results provide a mechanistic rationale and, together with the main outcomes of the TOFINGO study, support initiation of fingolimod within 8 weeks of natalizumab discontinuation.ClinicalTrials.gov identifier:NCT01499667.

Abstract 4077: Adipose-derived stem cells after Paclitaxel treatment demonstrate decreased function and suppression of breast cancer cell viability

Abstract Background: Following neoadjuvant or adjuvant chemotherapy for breast cancer resections, poor soft tissue wound healing can pose a major clinical challenge. Adipose-derived stem cells (ASCs) in clinically applied stromal vascular fractions are considered to play a positive role in wound healing (SVF) as evidenced by their increasing use in breast reconstructive procedures. Our study evaluated the potential confounding effects of Paclitaxel (PTX) on ASC proliferation and differentiation capacities and if PTX treated ASCs may subsequently influence tumor cell viability. Methods: IRB-approved human ASCs were isolated and treated with PTX at different concentrations. Proliferation, cell viability, and cell migration rates were measured by growth curves, MTT assays and scratch assays. ASCs were cultured in derivative-specific differentiation media with or without PTX (1uM) for 3 weeks. Adipogenic, osteogenic and endothelial differentiation levels were measured by quantitative RT-PCR, Oil-Red-O, Alizarin Red staining and cord formation on Matrigel, respectively. MDA-MB-231 (breast cancer cell line) and ASCs were co-cultured at equal cell numbers. ASC conditional media were collected every 3 days from ASC no-PTX treatment and PTX treated - washout groups. Results: PTX decreased proliferation of ASCs in a dose- and time-dependent manner. PTX treatment down-regulated the capacity of ASCs for adipogenic, osteogenic and endothelial differentiation (p<0.05 vs. control). In comparison with controls, PTX treated ASCs had decreased cell migration (relative scratch width, p<0.05 vs. control). Following cessation and washout of PTX after 8 days, treated ASCs exhibited cell growth and differentiating ability, but their capacity was suppressed compared to controls. The cumulative population doubling time was longer for cells co-cultured with PTX treated-washout ASC conditional media. (p<0.01 vs. no-PTX treatment group). Total cellular mRNA expressions of epithelial cell adhesion molecule (EPCAM), synuclein gamma (breast cancer specific protein 1; SNCG) and tumor necrosis factor alpha (TNF-α) were down-regulated in MDA-MB-231 cells when co-cultured directly with PTX treated ASC or indirectly with PTX treated-washout ASC conditional media (p<0.05 vs. control). In cells co-cultured for 3 days, VEGF proteins released in conditioned medium were decreased compared with single cultures (p<0.01 vs. control). Conclusion: Our results indicate that: 1) PTX inhibited ASC proliferation and decreased the multipotency differentiating capacity; and 2) Direct contact or treatment with conditional media from PTX treated ASCs suppressed tumor cell growth. In evaluating autologous ASCs for reconstructive procedures, this study may provide insight into poor soft tissue wound healing immediately following the removal of chemotherapy as well as decreased recurrence in breast cancer patients. Citation Format: William M. Harris, Michael Plastini, Telisha Ortiz, Nikolas Kappy, Jefferson Benites, Shaohua Chang, A. Lelani Fahey, Martha S. Matthews, Alexandre Hageboutros, Jeffrey P. Carpenter, Spencer Brown, Ping Zhang. Adipose-derived stem cells after Paclitaxel treatment demonstrate decreased function and suppression of breast cancer cell viability. [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 4077. doi:10.1158/1538-7445.AM2015-4077

Analysis of outcomes of intra-operative rectal washout in patients with rectal cancer during anterior resection

Local recurrence is a major setback for patients with rectal cancer after surgery. Implantation of exfoliated malignant cells has been considered as a possible mechanism of tumor recurrence. Although still controversial, intra-operative rectal washout has been described as a method to remove exfoliated malignant cells and reduce recurrence rates. The current study was designed to assess the differences in outcomes of patients with or without rectal washout. Data of 144 patients with rectal cancer undergoing anterior resection between 2006 and 2007 were reviewed in our study. 69 patients received rectal washout with approximately 1,500 ml physiological saline before anastomosis, and 75 patients received no-washout. The washout samples were sent for cytological examination. The incidence of recurrence and 5-year disease-free survival rates of the two groups were compared. Three patients had local recurrence in the washout group compared with 5 patients with no-washout during a median follow-up of 48 months (p = 0.721). The 5-year disease-free survival rate was 79.7 and 74.7 % in washout and no-washout groups, respectively (p = 0.553). Our results demonstrated that exfoliated malignant cells were ubiquitous in the rectal lumen during anterior resection. The difference of prognosis was not statistically significant, though trended toward being improved in the washout group. A large-scale research with longer follow-up time is required to draw a significant conclusion.

Washout policies in long‐term indwelling urinary catheterization in adults: A short version cochrane review

People requiring long-term bladder draining with an indwelling catheter can experience catheter blockage. Regimens involving different solutions can be used to wash out catheters with the aim of preventing blockage. To determine if certain washout regimens (including no washout) are better than others in terms of effectiveness, acceptability, complications, quality of life, and economics for the management of long-term indwelling urinary catheters in adults. We searched the Cochrane Incontinence Group Specialized Trials Register (searched April 30, 2009), MEDLINE (January 1966 to April 2009), MEDLINE In-Process (April 30, 2009), EMBASE (January 1980 to April 2009), and CINAHL (December 1981 to April 2009). Additionally, we examined all reference lists of identified trials and contacted manufacturers and researchers in the field. All randomized and quasi-randomized trials comparing catheter washout policies (e.g., washout vs. no washout, different washout solutions, frequency, duration, volume, concentration, method of administration) in adults (16 years and above) in any setting (i.e., hospital, nursing/residential home, community) with an indwelling urethral or suprapubic catheter in place for more than 28 days. Data were extracted by three reviewers independently and compared. Disagreements were resolved by discussion. Data were processed as described in the Cochrane Handbook. If the trial data were not fully reported, clarification was sought from the authors. For categorical outcomes, the numbers reporting an outcome were related to the numbers at risk in each group to derive a risk ratio (RR). For continuous outcomes, means, and standard deviations were used to derive weighted mean differences (WMD). No meta-analysis of study results was possible. Five trials met the inclusion criteria involving 242 patients (132 completed) in two cross-over and three parallel-group randomized controlled trials. Only three of the eight comparisons pre-stated in the review protocol were addressed in these trials. Some trials addressed more than one comparison (e.g., washout vs. no washout and one type of washout solution vs. another). The analyses reported for the two cross-over trials were inappropriate as they were based on differences between groups rather than differences within individuals receiving sequential interventions. Two parallel-group trials had limited value: one combined results for suprapubic and urethral catheters and one had data on only four participants. Only one trial was free of significant methodological limitations, but its sample size was small. Three trials compared no washout with one or more washout solution (saline or acidic solutions) and authors tended to conclude no difference in clinical outcomes between washout and no washout. In the one trial which had data of sufficient quality to allow interpretation, no difference was detected between washout and no washout groups in the rate of symptomatic urinary tract infection or time to first catheter change. Three trials compared different types of solution: saline versus acidic solutions (two trials); saline versus acidic solution versus antibiotic solution (one trial). Authors tended to report no difference between different washout solutions but the data were too few to support their conclusions. The one trial which warranted consideration concluded no difference between saline and an acidic solution in terms of symptomatic urinary tract infections or time to first catheter change. The data from five trials comparing differing washout policies were sparse and trials were generally of poor quality or poorly reported. The evidence was too scant to conclude whether or not washouts were beneficial. Further rigorous, high quality trials with adequate power to detect any benefit from washout rather than no washout being performed are required in the first instance. After that, trials comparing different washout solutions, washout volumes, frequencies/timings, and routes of administration are needed.

Methimazole-induced hypothyroidism inhibits the panic-like behaviors produced by electrical stimulation of dorsal periaqueductal gray matter of rats

Conflicting clinical data on the relationship of panic disorder and thyroid diseases illustrate the need for a simpler approach using animal models. Defensive behaviors evoked by electrical or chemical stimulation of dorsal periaqueductal gray matter (DPAG) have been proposed as a model of panic attack. Therefore, the present study examined the effects of the anti-thyroid agent methimazole (MTZ) either on the panic-like behaviors induced by electrical stimulation of DPAG or the anxiety-like behaviors of rats exposed to the elevated plus-maze (EPM). Male Wistar rats bearing electrodes in the DPAG were stimulated with stepwise increased currents. Rats which displayed galloping at intensities below 60muA were retested following 5- and 10-day treatments with MTZ (0.6mg/kg/day, i.p.) or 10- and 15-day washout periods. MTZ effects on EPM performance were assessed in separate groups. MTZ-treated groups were compared to saline-treated controls. In other experiments, rats were similarly treated with MTZ and the blood was collected for hormone assays. The 10-day treatment with MTZ produced marked increases in the thresholds of exophthalmus (65%), immobility (75%), trotting (63%), galloping (56%), jumping (47%), defecation (114%) and micturition (85%). Effects outlasted the drug discontinuation. In contrast, MTZ had variable effects in the EPM, significantly increasing the open-arm exploration in 5-day treated and 10-day washout groups. Biochemical data revealed a small but significant decrease (13%) in free thyroxine in MTZ-treated groups. Although not significant, thyrotrophin levels showed a 111% increase following the 10-day treatment with MTZ. Selective attenuation by MTZ of DPAG-evoked defensive behaviors supports attenuation of panic attacks in hypothyroidism.

Hypocapnia prolongs bradycardia induced by bupivacaine or levobupivacaine in isolated rat hearts

Systemic alkalinization is recommended for resuscitation from local anesthetic-induced cardiotoxicity. It has been suggested that inducing hypocapnic alkalosis, prior to exposure to toxic concentrations of local anesthetics, may minimize cardiotoxicity. However, it remains unclear whether inducing severe hypocapnic alkalosis after administration of local anesthetics will minimize the duration of bradycardia. We used isolated rat hearts to investigate the effects of hypocapnic alkalosis on heart rate (HR) recovery from bupivacaine or levobupivacaine-induced bradycardia. We measured the time required for the HR in 24 isolated rat hearts, respectively, to attain 90% of the baseline HR (recovery time) following bradycardia induced by 1 microg x mL(-1) and 10 microg x mL(-1) concentrations of either bupivacaine or levobupivacaine. Normal pH perfusate (bupivacaine or levobupivacaine with normal pH washout groups) or severe hypocapnic alkalosis perfusate (bupivacaine or levobupivacaine with hypocapnic alkalosis washout groups) were reperfused after exposure to the local anesthetics. Severe hypocapnic alkalosis prolonged the recovery time from 273 +/- 122 sec, at the 1 microg x mL(-1) bupivacaine concentration with normal pH washout, to 1203 +/- 540 sec, in the bupivacaine with hypocapnic alkalosis washout (p = 0.029). Severe hypocapnic alkalosis also prolonged the recovery time from 1153 +/- 644 sec, at a 10 microg x mL(-1) bupivacaine concentration in the normal pH washout group, to 2065 +/- 617 sec, in the bupivacaine with hypocapnic alkalosis washout group (p = 0.032). With levobupivacaine 10 microg x mL(-1) in the normal pH washout group, HR recovery time increased from 863 +/- 186 sec to 1565 +/- 567 sec, compared to the hypocapnic alkalosis washout group (p = 0.045). Severe hypocapnic alkalosis prolonged the recovery time from bupivacaine or levobupivacaine-induced bradycardia in isolated rat hearts. When bradycardia occurs after intravascular bupivacaine or levobupivacaine administration, maintenance of normocapnia may minimize the duration of bradycardia.

Reversal of Prilocaine Epidural Anesthesia Using Epidural Saline or Ringer’s Lactate Washout

Several investigators have described the phenomena of epidural saline washout using bolus injections. This study was designed to determine whether epidural block could be reversed more effectively by infusion of crystalloid solutions via the epidural catheter. One hundred male patients scheduled for outpatient surgery were enrolled in this study. After 30 min of 2% prilocaine epidural anesthesia, patients were randomly assigned to receive 45 mL of study solution as follows: (1) normal saline bolus (group NSB); (2) Ringer's lactate bolus (group RLB); (3) normal saline infusion (group NSI); (4) Ringer's lactate infusion (group RLI). Patients in the control group received no washout fluid. Motor, sensory blockade and side effects were compared among 5 groups. Ambulation time is defined as the recovery time. In the control group, ambulation time (139 +/- 15 min) was significantly longer than in the washout groups (NSB 90 +/- 10, RLB 88 +/- 10, NSI 85 +/- 8, RLI 91 +/- 6 minutes) (P < .001). Two-segment sensory regression time in the control group (86 +/- 15 min) was significantly longer than in groups NSB, RLB, NSI and RLI (55 +/- 8, 51 +/- 4, 58 +/- 8, and 53 +/- 10 minutes, respectively) (P < .001). We concluded that a more rapid recovery of motor and sensory blockade in patients undergoing epidural anesthesia may be achieved by the use of an epidural washout with either bolus or infusion of 45 mL normal saline or Ringer's lactate.

Maternal caffeine intake impairs MK-801-induced hyperlocomotion in young rats

Here we have investigated the effects of maternal caffeine intake (1 g/l) on MK-801-induced hyperlocomotion in rat pups. Animals submitted to caffeine treatment during the gestational and lactational period were separated in two groups: caffeine-treated group (up to 21 days old) and washout group (caffeine treatment up to 7 days old). MK-801 (0.25 mg/kg, i.p.) promoted hyperlocomotion in control rats, but this stimulatory effect was significantly decreased in caffeine-treated and washout groups. The permanent effect after caffeine withdrawal suggests durable or adaptive changes during neurodevelopment, mainly on adenosine receptors or neurotransmitter systems modulated by adenosine, such as the glutamatergic system.

Myocardial protection by coronary washout during global ischemic cardiac arrest

The effect of intermittent coronary washout (WO) during global ischemic cardiac arrest (ICA) was evaluated in isolated blood perfused dog hearts undergoing 90 min normothermic ICA and 90 min reperfusion. WO consisted of infusion of 100 ml normothermic dog plasma at 100 mm Hg every 10 min during ICA. Systolic and diastolic pressures were measured at constant volume with a left ventricular balloon. Coronary blood flow (CBF) was measured, and transmyocardial oxygen, lactate, and glucose differences were calculated. Adenosine triphosphate (ATP), creatine phosphate (CP), calcium (Ca 2+), glycogen, and water content were measured from left ventricular biopsies. During 90 min of WO, hearts extracted glucose (15 ± 4 mg/g dry wt LV) and lost lactate (90 ± 5 μmole/g dry wt LV). ATP and CP were significantly depressed at arrest in both groups although WO resulted in significantly higher levels of ATP and CP. CP returned to control levels at 90 min reperfusion in the WO group but remained depressed in the ICA group. Calcium accumulation was greater in hearts with ICA. With reperfusion, both ICA and WO groups demonstrated early hyperemia and rapid lactate washout. There were no differences in total CBF, AVO 2, MOV 2, and myocardial glycogen or water content. Systolic performance was equally depressed in both groups at 90 min of reperfusion. Diastolic compliance, while still impaired, was better preserved in WO when measured in the arrested heart at 90 min ICA ( P < 0.01) and in the contracting heart after 90 min reperfusion ( P < 0.02). Normothermic coronary washout during 90 min of normothermic ischemic arrest without cardioplegic agents produces improved ventricular compliance and energy metabolism, and reduces myocardial calcium accumulation.