BSO Group Research Articles

Working and episodic memory trajectories over time following bilateral oophorectomy in early midlife women

AbstractBackgroundEarly ovarian hormone deprivation due to bilateral salpingo‐oophorectomy (BSO; ovarian and fallopian tube removal) is a risk for late‐life AD, but estradiol replacement therapy (ERT) may be protective (Rocca et al., 2007). Working and episodic memory decrements have been observed without ERT soon after surgery (Sherwin et al., 1988) and on average 5 years post‐BSO in midlife compared to premenopausal controls (Gervais et al., 2020). Unknown is whether memory continues to decrease over time in early midlife, a critical period for understanding cognitive trajectories that presage AD. We investigated memory changes in early midlife over time post‐BSO, and whether those taking ERT had different trajectories than those without.MethodParticipants with BSO taking ERT (BSO+ERT: M age = 44.5y; n = 71) were compared to those not taking ERT (BSO: M age = 43.2y; n = 55), recruited from Toronto and Montreal, Canada, and Linköping, Sweden, and completed up to four yearly visits. Episodic memory was evaluated using Logical Memory Task (LM) percent forgetting and Rey Auditory Verbal Learning Test (RAVLT) total learning. Working memory was assessed using Spatial Working Memory Task (SPWM) trial 1 errors, Digit Ordering Task (DOT) errors, and Digit Span Backward Task (DSB) span length. Multilevel models were used to examine whether ERT influenced memory over time post‐BSO and disaggregate within‐ and between‐person effects. Estimated marginal means (Aiken and West, 1991) were calculated to delineate significant interactions.ResultThere were no significant changes over time post‐BSO on LM or RAVLT. The DSB showed significant within‐ and between‐person effects; the BSO group showed decreases in span length over time post‐BSO, while the BSO+ERT group showed increases and had significantly longer span lengths at seven years post‐BSO and beyond (Fig.1A&B). The SPWM Task showed a significant within‐person effect; the BSO group had stable errors over time post‐BSO, while the BSO+ERT group had working memory improvements and made fewer errors (Fig.1C).ConclusionFindings suggest that immediate verbal episodic memory declines we observed previously may stabilize. They also reveal that without ERT, most women have working memory declines post‐BSO. These data support ERT administration post‐BSO for working memory preservation. If ERT is contraindicated, cognitive training or lifestyle interventions may mitigate working memory declines.

Women’s Brain Health: Midlife Ovarian Removal Affects Associative Memory

Women with early bilateral salpingo-oophorectomy (BSO; removal of ovaries and fallopian tubes) have greater Alzheimer’s disease (AD) risk than women in spontaneous/natural menopause (SM), but early biomarkers of this risk are not well-characterized. Considering associative memory deficits may presage preclinical AD, we wondered if one of the earliest changes might be in associative memory and whether younger women with BSO had changes similar to those observed in SM. Women with BSO (with and without 17β-estradiol replacement therapy (ERT)), their age-matched premenopausal controls (AMC), and older women in SM completed a functional magnetic resonance imaging face-name associative memory task shown to predict early AD. Brain activation during encoding was compared between groups: AMC (n=25), BSO no ERT (BSO; n=15), BSO+ERT (n=16), and SM without hormone therapy (n=16). Region-of-interest analyses revealed AMC did not contribute to functional group differences. BSO+ERT had higher hippocampal activation than BSO and SM. This hippocampal activation correlated positively with urinary metabolite levels of 17β-estradiol. Multivariate partial least squares analyses showed BSO+ERT had a different network-level activation pattern than BSO and SM. Thus, despite being approximately 10 years younger, women with BSO without ERT had similar brain function to those with SM, suggesting early 17β-estradiol loss may lead to an altered functional brain phenotype which could influence late-life AD risk, making face-name encoding a potential biomarker for midlife women with increased AD risk. Despite similarities in activation, BSO and SM groups showed opposite within-hippocampus connectivity, suggesting menopause type is an important consideration when assessing brain function.

Do Barn Swallow nestlings incur an oxidative cost of begging?

Some theoretical models predict that nestling begging must be a costly activity to transmit reliable information to parents about offspring hunger. One candidate cost is oxidative stress, which could impair growth or immune function. This hypothesis predicts that nestlings in a poor oxidative status should pay higher costs for a given amount of begging, hence beg at lower rates for a similar degree of hunger. To test this prediction, we experimentally induced a transient alteration of the oxidative status of Barn Swallow (Hirundo rustica) nestlings by administering them either a single dose of a pro-oxidant substance (BSO) or an antioxidant (vitamin E), and recorded their begging behaviour. Chicks treated with vitamin E increased their begging rates in the hours following treatment. However, and contrary to prediction, BSO nestlings did not decrease begging compared to controls. These inconsistent results are difficult to explain by the hypothesis that begging performance is mediated by the oxidative status of nestlings because chicks with an impaired oxidative status (the BSO group) begged the same as controls. Alternatively, vitamin E may alleviate the inhibitory effects on begging of stress responses mediated by glucocorticoids. This study suggests that oxidative costs of begging are minor or absent in this species. Future studies should consider alternative explanations for an association between oxidative status, antioxidant supplementation and begging behaviour, particularly those involving glucocorticoids.

GPP05 Presentation Time: 8:40 AM: Comparing Long Term Quality of Life Across Different Uterine Cancer Treatment Modalities

<h3>Objectives</h3> Uterine cancer is the most common gynecologic malignancy in the United States; however, little is understood of long-term sequelae of adjuvant radiotherapy treatment. The purpose of this study was to compare quality of life (QoL) in patients who have undergone surgery with or without adjuvant radiotherapy for uterine cancer. <h3>Materials and Methods</h3> This is a cross-sectional cohort study that examined women treated for uterine cancer at a tertiary cancer center from 2006-2018. Participants included those who underwent hysterectomy/bilateral salpino-oophorectemy alone (BSO), with brachytherapy (BT), or with external bean radiation therapy (EBRT). A non-cancer cohort of women who underwent a hysterectomy/BSO for benign indications was also identified (NC). To compare QoL we utilized the FACT-En, a validated 27-question survey used to assess QoL in endometrial cancer patients. The survey consists of 5 well-being subscales: physical, social, emotional, functional, and an endometrial-cancer specific subscale. Responses are on a 5-point Likert scale, with a higher score indicating a better quality of life. Maximum score is 172 for the entire survey. Demographic and other treatment variables were obtained from medical records. Subscales and survey totals were compared across cohorts using ANOVA tests. <h3>Results</h3> 282 women completed the FACT-En survey (BSO 59, BT 70, EBRT 93, NC 65). Median time from surgery to completion of survey was 6.7 years. Women in the NC cohort were significantly younger at survey completion compared to the other cohorts (mean age 63 [std 8.2] NC, 66 [9.6] HS, 68 [8.4] BT, 67 [10.8] EBRT; p<0.001). Mean total FACT-En score for the entire cohort was 114.2 [22.1]. Overall QoL was significantly different between cohorts, with patients in the EBRT cohort reporting the lowest QoL (mean 139.4 [21.6]), followed by the NC cohort (142.1 [26.5]), BSO cohort (146.1 [20.1]), and the BT cohort reporting highest total score (150.6 [18.2], p=0.006). All subscales were significantly different (all p<0.05) across cohorts aside from social well-being subscale. In the physical domain, the EBRT cohort had the lowest average score (23.3 [5.0]), followed by the NC cohort (25.3 [4.2]), the BT cohort (25.0 [3.8]), with the BSO group reporting the highest scores (25.4 [3.9], p=0.001). The NC cohort reported the lowest scores in the endometrial symptom-specific subscale, with a mean score of 53.3 [10.5]. Among patients who had undergone cancer treatment, the EBRT cohort reported the worst average endometrial-specific QoL (53.5 [8.6]), while again the BT group reported the highest score (57.5 [6.1], p=0.007). <h3>Conclusion</h3> QoL differences between treatment modalities for uterine cancer remain years after treatment. In women with endometrial cancer who require adjuvant therapy, vaginal brachytherapy does not appear to have any long-term effects of reported QOL.

Depletion of glutathione induced apoptosis and oxidative stress via the activation of TRPM2 channels in the microglia cells with Alzheimer’ disease model

Alzheimer’s disease is a common neurodegenerative disease. Microglia induces oxidative stress in the brain for engulfing bacteria and viruses. The accumulating data indicate that oxidative stress and apoptosis are two main actors for the induction of microglia activation-induced Alzheimer’s Disease. Oxidative stress is one of many triggers that activate the transient receptor potential melastatin 2 (TRPM2) channel. Glutathione (GSH) is a main cytosolic antioxidant in the mammalian cells. The GSH depletion via the activation of TRPM2 induces oxidative stress and apoptosis in neuronal cells. It has not yet been researched how GSH depletion via activation of TRPM2 affects oxidative stress and apoptosis in microglial cells with the Alzheimer's disease model. The BV2 cells divided into 5 groups as control, buthionine sulphoximine (BSO and 0.5 mM for 6 h), amyloid beta (1 uM for 72 h), amyloid beta+BSO, and amyloid beta+BSO+GSH (10 mM for 2 h). In the BSO group, the levels of apoptosis, mitochondrial membrane potential, cytosolic free oxygen reactive species (cyROS), caspase (Casps) -3, Casps -8, and Casps -9 were increased as compared to the control group, although cell viability level was decreased. The expression levels of TRPM2, Casps -3, Casps -9, Bax, Bcl-2, and PARP-1 were also increased in the BSO group. In addition, their levels were further increased in the amyloid beta and BSO+amyloid beta groups as compared to the BSO group. However, the changes were modulated in the BSO+amyloid beta+GSH group by the incubation of GSH. In conclusion, the depletion of GSH increased apoptosis and cyROS levels via activation of caspases and TRPM2 in the amyloid beta-induced microglia cells. The treatment of GSH may be a potential target on the apoptosis and oxidative stress in the amyloid beta-induced microglia cells.

OPEC study: An international multicenter study of ovarian preservation in endometrial cancers.

To evaluate the feasibility and oncological safety of ovarian preservation in early stage endometrial adenocarcinoma (EC) patients aged 40 and below. A total of 11 institutions from eight countries participated in the study. 169 of 5898 patients aged ≤40 years were eligible for the study. Patients with EC treated between March 2007 and January 2019 were retrospectively assessed. The median duration of follow-up after EC diagnosis was 59 months (4-187). Among 169 participants, ovarian preservation surgery (OPS) was performed in 54 (31.9%), and BSO was performed in 115 (68.1%) patients. Although patients younger than 30 years of age were more likely to have OPS than patients aged 30 to 40 years (20.4% vs. 9.6%, P =0.021), there was no significant difference by the mean age. There were no other relevant baseline differences between OPS and BSO groups. The Kaplan-Meier analysis revealed no difference in either the overall survival (P =0.955) or recurrence-free survival (P =0.068) among patients who underwent OPS, and BSO. OPS appears to be safe without having any adverse impact on survival in women aged ≤40 years with FIGO Stage I EC.

Disturbed sleep and reduced hippocampal integrity at midlife: Implications for AD risk in women with early surgical menopause

AbstractBackgroundWomen bear the greatest burden of Alzheimer’s disease (AD), and early (&lt;45y) ovarian hormone deprivation via surgical menopause further increases risk (Rocca et al., 2007). Given that sleep disturbance is implicated in AD progression (Lim et al., 2013), an important area of investigation is determining whether younger women with early surgical menopause demonstrate disturbed sleep at midlife, and whether this may confer additional risks by contributing to changes in hippocampal integrity. Thus, we investigated whether middle‐aged women with bilateral salpingo‐oophorectomy not taking hormone therapy demonstrate sleep disturbance, and reduced hippocampal volume and memory. Additionally, we assessed whether sleep and hippocampal volume predict memory performance.MethodWomen with an early BSO that were either taking estradiol‐based hormone therapy (BSO+ET: age=43y; n=22), or not taking ET (BSO: age=46y; n=18) were recruited to the study, and were compared to age‐matched premenopausal (AMC: n=24) and spontaneously postmenopausal women who were ∼10y older (SM: 55y; n=20). Sleep was assessed via portable polysomnography (Vitaport‐5/REMbo‐234, Temec Technologies) for 1‐3 nights, and sleep staging was acquired using automated scoring (Z3score, Neurobit technologies). High‐resolution T2‐weighted scans were acquired using a Siemens 3T Prisma scanner, and hippocampal subfields (CA1, Subiculum, combined region: DGCA23), were manually segmented (Olsen et al., 2017) in FSLView. The interference match‐to‐sample (IMTS) task was used to asses and recognition memory for scenes (Watson et al., 2013).ResultBoth hormone‐deprived groups demonstrated increased sleep latency (BSOvsAMC: p&lt;.05, d= 0.74; SMvsAMC: d= 0.69) and decreased sleep efficiency (BSOvsAMC: p&lt;.05, d= 0.79; SMvsAMC: p&lt;.05, d= 0.67). Only the BSO group demonstrated reduced volume in the anterior CA1 and DGCA23 relative to AMC (p&lt;.05, d= 0.96‐1.06) and BSO+ET (p&lt;.05, d= 0.67‐0.84). Additionally, the BSO demonstrated reduced efficiency (accuracy/RTcorrect) on scene recognition memory (p&lt;.05, d=0.70‐0.86). Multiple regression analyses show that DGCA23 volume and group membership predict scene recognition memory performance.ConclusionWhile both hormone‐deprived groups demonstrated disturbed sleep, women with early BSO also showed reduced hippocampal integrity, despite being ∼10 years younger than SM women. These findings are consistent with the elevated AD risk among women with BSO. It remains to be seen whether sleep disturbance at a younger age exacerbates risk.

A Hospital Based Cross-Sectional Study to Evaluate Ovarian Cancer Risk after Salpingectomy

Objective: The main objective of this hospital based cross-sectional study was to evaluate ovarian cancer risk after salpingectomy. Material and Methods: This hospital based cross-sectional observational study was carried out over a period of 1 year (Oct 2019 – Oct 2020) conducted at SS Hospital, Bihar. By descriptive statistics & Pearson Chi square test using SPSS software the collected data were analysed. Results: 257 patients were included in salpingectomy group, whereas 312 were under Hysterectomy and BSO group. Mean age at entry was 35.7± 8.9 years at Salpingectomy group whereas the same was 37.9 ±4.8 years with Hysterectomy and BSO group. When compared with the Hysterectomy and BSO population for ovarian cancer and among women with previous salpingectomy there was a statistically significantly lower risk observed. A further 50% decrease in risk of ovarian cancer was associated with Bilateral salpingectomy compared with the unilateral procedure (unilateral: HR = 0.73, 95% CI = 0.57 to 0.92; bilateral salpingectomy: HR = 0.36, 95% CI = 0.17 to 0.75). Conclusion: Reduced risk of ovarian cancer is associated with salpingectomy on benign indication. This data also in line with the concept that in the fallopian tube ovarian cancer arises at a substantial fraction. Our results suggest that in the general population to reduce ovarian cancer risk an effective measure is the removal of the fallopian tubes by itself.

EFFECT OF SUPPLEMENTATION OF AROMATIC PLANTS OILS ON IMMUNITY, UDDER HEALTH AND MILK PRODUCTION OF FRIESIAN COWS

Eight weeks prior to expected calving date, 32 Friesian cows were assigned according to parity to four homogenous groups (8 animals each). Cows were fed a basal ration alone as a control group 1 (CON G1), or basal ration supplemented with 25 mg black seed oil per kg body weight, (BSO G2), 11 mg chamomile flower oil (CFO G3), or 25 mg oregano leaves oil (OLO G4). Biochemical and hematological parameters were measured and there were within the normal ranges of cattle. Relative to the control group, aromatic plant oils significantly reduced plasma lipids and increased plasma concentration of IgG. Feed intake was similar among groups. Economic evaluations were in favor of BSO group. BSO significantly (P&lt;0.05) improved daily milk yield (DMY), fat and protein, but reduced lactose content of milk. CFO increased (P&lt;0.05) daily milk yield. Only fat-corrected DMY was increased with OLO supplementation. The three supplements reduced (P&lt;0.05) milk electric conductivity (EC) and somatic cell count (SCC). Overall, our results indicated that supplementing diets of Friesian cows with aromatic plant oils, particularly black seed oil, improved milk yield, udder health, and some immune parameters.Key words: aromatic oils; cows; blood parameters; milk production and immunity

EFFECT OF DIET SUPPLEMENTED WITH PUMPKIN (CUCURBITA MOSCHATA) AND BLACK SEED (NIGELLA ATIVA) OILS ON PERFORMANCE OF RABBITS: 3- PRODUCTIVE AND REPRODUCTIVE TRAITS, PUBERTY, SEXUAL ACTIVITY AND SEMEN CHARACTERISTICS OF NEW ZEALAND WHITE MALE RABBITS

Forty New Zealand White (NZW) rabbit males aged 12 weeks and weighing 1540.80±22.16 g were equally and randomly divided into four groups (10 in each). All groups were fed a commercial pelleted diet and the first group was served as a control, while diet of the second group was supplemented with 5 g pumpkin seed oil/kg diet (PSO), the third group received 5 g black seed oil/kg diet (BSO), however the fourth group received 5 g from both PSO and BSO/kg diet (PSO+BSO). Results showed that male rabbits in PSO+BSO group showed the heaviest (P<0.05) live body weight (LBW), total weight gain (TWG), average daily gain (ADG), feed conversion ratio and performance index, followed by those in PSO or BSO groups, while males in control group had the light values. Male rabbits received diet supplemented with PSO+BSO combination reached puberty significantly (P<0.05) faster compared with control group, while PSO or BSO groups were intermediate with insignificant differences. Moreover, buck rabbits in PSO+BSO group showed the highest (P<0.05) scrotal circumference, testicular index, weights of testes, epididymis and sexual accessory glands (included seminal vesicles, prostate and the paired bulbourethral (Cowper’s gland), plasma testosterone concentration at puberty and the short reaction time and latency period, followed by those in either PSO or BSO groups, however bucks in control group had the lowest values. Semen volume and gel, pH value and semen color score were significantly (P<0.05) higher in PSO+BSO group than that in control group, PSO or BSO groups had intermediate values with insignificant differences. Semen color tended to be milky in PSO+BSO and BSO groups and heavy watery in PSO and control groups. All semen characteristics differ significantly (P<0.05) among the different collection weeks with a tendency to improve with advancing age. The PSO+BSO group showed highly (P<0.001) significant of percentages of sperm motility, livability, sperm cell concentration and the lowest sperm abnormality percentage, followed by PSO or BSO groups, however the control group had the lowest values. All sperm characteristics were significantly (P<0.05) differ among the different collection weeks and improved with age progress. Bucks in PSO+BSO group recorded the highly (P<0.001) significant of total count, total motile, total live and total normal per ejaculate, followed by those in PSO or BSO groups, however, the lowest in control group. Also, all counts of spermatozoa were significantly (P<0.05) differ among the different collection weeks and tended to increase with age.

오메가-3 지방산 함유 복분자종자유에 의한 고지방식이 유도 고지혈증 마우스의 혈중 중성지방 및 콜레스테롤 감소 효과

To investigate the effect of n-3 fatty acid from Bokbunja (Rubus coreanus Miq.) seed oil (BSO), we examined improvement of plasma triglycerides and cholesterol in vivo. Five-week-old ICR mice were divided into five groups of six mice each; Control, high fat diet (HFD) control (negative control), salmon oil control (positive control, HFD+commercial n-3 fatty acid), and BSO experimental groups (HFD+1 g/60 kg BW/d, HFD+2 g/60 kg BW/d). After 4 weeks of BSO treatment, we measured serum triglyceride and cholesterol levels. The levels of low-density lipoprotein/very-low-density lipoprotein-cholesterol, high-density lipoprotein-cholesterol, and total cholesterol were significantly (P<0.05) reduced in the group fed BSO at 2 g/60 kg BW/d compared to the negative control. Levels of triglycerides, which are similar to cholesterol, were also significantly (P<0.05) reduced in the same group. To investigate further, we tested blood coagulation parameters. Prothrombin time (PT) and activated partial thromboplastin time (aPTT) were not significantly different among the five groups according to BSO. However, the 2 g/60 kg BW/d BSO group treated with PT and aPTT showed a tendency to live longer than the negative control. Taken together, BSO might improve blood homeostasis mediated via hypo-lipidemic and anti-coagulation activities.

Impaired glutathione redox system paradoxically suppresses angiotensin II-induced vascular remodeling.

BackgroundAngiotensin II (AII) plays a central role in vascular remodeling via oxidative stress. However, the interaction between AII and reduced glutathione (GSH) redox status in cardiovascular remodeling remains unknown.Methods In vivo: The cuff-induced vascular injury model was applied to Sprague Dawley rats. Then we administered saline or a GSH inhibitor, buthionine sulfoximine (BSO, 30 mmol/L in drinking water) for a week, subsequently administered 4 more weeks by osmotic pump with saline or AII (200 ng/kg/minute) to the rats. In vitro: Incorporation of bromodeoxyuridine (BrdU) was measured to determine DNA synthesis in cultured rat vascular smooth muscle cells (VSMCs).ResultsBSO reduced whole blood GSH levels. Systolic blood pressure was increased up to 215±4 mmHg by AII at 4 weeks (p<0.01), which was not affected by BSO. Superoxide production in vascular wall was increased by AII and BSO alone, and was markedly enhanced by AII+BSO. The left ventricular weight to body weight ratio was significantly increased in AII and AII+BSO as compared to controls (2.52±0.08, 2.50±0.09 and 2.10±0.07 mg/g respectively, p<0.05). Surprisingly, the co-treatment of BSO totally abolished these morphological changes. Although the vascular circumferential wall stress was well compensated in AII, significantly increased in AII+BSO. The anti-single-stranded DNA staining revealed increasing apoptotic cells in the neointima of injured arteries in BSO groups. BrdU incorporation in cultured VSMCs with AII was increased dose-dependently. Furthermore it was totally abolished by BSO and was reversed by GSH monoethyl ester.ConclusionsWe demonstrated that a vast oxidative stress in impaired GSH redox system totally abolished AII-induced vascular, not cardiac remodeling via enhancement of apoptosis in the neointima and suppression of cell growth in the media. The drastic suppression of remodeling may result in fragile vasculature intolerable to mechanical stress by AII.

Maternal omega-3 fatty acid supplementation on vitamin B12 rich diet improves brain omega-3 fatty acids, neurotrophins and cognition in the Wistar rat offspring

The consequences of wide spread vegetarianism due to low vitamin B12 on brain development and functioning is gaining importance. However, there are no studies which have evaluated exclusively vitamin B12 supplementation during pregnancy on brain growth. A series of our animal studies have documented adverse effects of maternal micronutrient imbalance on brain neurotrophins and its amelioration by omega-3 fatty acids. Therefore, the present study investigated the effect of maternal supplementation with vitamin B12 alone and B12 plus omega-3 fatty acid on pup brain fatty acids and neurotrophins at birth and 3 mo of age. Pregnant Wistar rats and their male offspring were assigned to 3 dietary groups: Control (normal vitamin B12 (25 μg/kg), vitamin B12 supplemented (BS) (50 μg/kg), vitamin B12 supplemented with omega-3 fatty acid (BSO) till 3 month of age. Maternal vitamin B12 supplementation (BS) increased brain BDNF (protein and mRNA) and DHA levels in pups at birth and in the hippocampus at 3 month of age (BDNF only). These effects were further enhanced by omega-3 fatty acid supplementation to vitamin B12 supplemented group. The spatial memory performance was found to be enhanced in BSO group which was characterised by less number of errors in radial eight arm maze. Our results indicate that a combination of omega-3 fatty acid and vitamin B12 enriched diet may exert beneficial effects on synaptic plasticity and cognition, which may prove beneficial for mental health, particularly in preventing neurocognitive disorders.

Cardiomyocyte-derived mitochondrial superoxide causes myocardial electrical remodeling by downregulating potassium channels and related molecules.

This study was designed to investigate the role of a primary hyperoxidative stress in myocardial electrical remodeling using heterozygous heart/muscle-specific manganese superoxide dismutase-deficient (H/M-Sod2(+/-)) mice treated with L-buthionine-sulfoximine (BSO). Both H/M-Sod2(+/-)and wild-type (WT) mice were treated with intra-peritoneal BSO or saline for 7 days, and divided into 4 groups: H/M-Sod2(+/-)+BSO, WT+BSO, H/M-Sod2(+/-)control, and WT control. The ventricular effective refractory period (ERP) and the monophasic action potential duration (MAPD) were determined. Levels of oxidative stress, potassium channel-related molecules, and K(+)channel-interacting protein-2 (KChIP2) were also evaluated. The H/M-Sod2(+/-)+BSO group exhibited markedly prolonged MAPD20, MAPD90 and ERP in comparison with the other groups (MAPD20: 14 ± 1 vs. 11 ± 1 ms, MAPD90: 77 ± 7 vs. 58 ± 4 ms, ERP: 61 ± 6 vs. 41 ± 3 ms, H/M-Sod2(+/-)+BSO vs. WT control; P<0.05). Mitochondrial superoxide and hydrogen peroxide formation in the myocardium increased in the H/M-Sod2(+/-)+BSO group in comparison with the WT+BSO group (P<0.05). Real-time RT-PCR and Western blotting revealed that Kv4.2 expression was downregulated in both BSO-treated groups, whereas KChIP2 expression was downregulated only in the H/M-Sod2(+/-)+BSO group (P<0.05). BSO treatment caused hyperoxidative stress in the myocardium of H/M-Sod2(+/-)mice. Changes in the expression and function of potassium channels were considered to be involved in the mechanism of electrical remodeling in this model.

Evaluation of hepatic damage by reactive metabolites--with consideration of circadian variation of murine hepatic glutathione levels.

Generally, reactive metabolites are detoxified by conjugation with glutathione (GSH). A GSH-depleted model was prepared by administering L-buthionine-(S,R)-sulfoximine (BSO), which can be used to detect hepatic damage by reactive metabolites. However, BSO may cause adverse effects on other organs, such as renal damage by reactive metabolites because it depletes GSH in the whole body. The present study was designed to examine whether it was possible to specifically detect hepatic damage by reactive metabolites without reducing renal GSH levels by administering BSO in a time course when hepatic GSH levels are naturally reduced. Male BALB/c mice were administered reverse osmosis (RO) water or 20 mmol/l BSO in drinking water for 4 days. Subsequently, animals in the RO water group were orally administered 500 mg/kg acetaminophen (APAP) at 9:00 or 15:00 and in the BSO group at 9:00 for 4 days. As a result, severe hepatic damage and necrosis of the renal proximal tubules were observed in the BSO/APAP administration at 9:00 group, and all animals died on 1 or 2 days after APAP administration. Hepatic damage was clearly increased in the RO water/APAP administration at 15:00 group compared with the RO water/APAP administration at 9:00 group. However, renal damage and deaths were not observed. This BSO administration model may detect renal damage induced by reactive metabolites. Using an administration time course, whereby hepatic GSH levels were naturally reduced, hepatic damage by reactive metabolites can be detected without secondary renal effects.

Is pre-operative risk-assessment in laparoscopic treatment of presumed low-risk endometrial cancer effective?

In endometrial (pre)malignancy the pre-operative work-up is primarily based on the histopathological specimen obtained. Total laparoscopic hysterectomy with bilateral salpingo-oophorectomy (TLH + BSO) in presumed low-risk clinical stage I endometrioid endometrial carcinoma (EEC) or atypical hyperplasia (AH), is nowadays considered preferred and sufficient treatment in the Netherlands. To test the effectiveness of this pre-operative work-up, a retrospective cohort analysis was performed. Revised pre- and post-operative histopathology was compared and intra- and post-operative complications registered. In 116 consecutive patients with a pre-operative diagnosis of AH or presumed stage I, grade I or II EEC planned for TLH + BSO. In 24.1 % (28/116) revised endometrial histopathology was upgraded on the definitive hysterectomy specimen. In 3.5 % (4/116) upgrading to high-risk grade III endometrial cancer (EC) was observed. In 9.9 % (8/81) of EC cases a post-operative FIGO stage IG3, II, or III was diagnosed. The major and minor short-term complication rates of TLH + BSO were 12.1 and 7.8 %. In 13.8 % (16/116) of cases conversion to laparotomy was necessary, with a significant higher percentage of obese (68.8 %) patients in the conversion versus the successful TLH + BSO group (42 %). Clinical relevant inconsistency between pre- and post-operative histopathology or FIGO stage was observed in 9.9 % of EC cases. More extensive pre-operative risk analysis of presumed low-risk EEC may be indicated, especially for the morbid obese, harboring a substantial risk for conversion to laparotomy and complications.

In vivo inhibition of l-buthionine-( S,R)-sulfoximine-induced cataracts by a novel antioxidant, N-acetylcysteine amide

The effects of N-acetylcysteine amide (NACA), a free radical scavenger, on cataract development were evaluated in Wistar rat pups. Cataract formation was induced in these animals with an intraperitoneal injection of a glutathione (GSH) synthesis inhibitor, l-buthionine-( S,R)-sulfoximine (BSO). To assess whether NACA has a significant impact on BSO-induced cataracts, the rats were divided into four groups: (1) control, (2) BSO only, (3) NACA only, and (4) NACA + BSO. The control group received only saline ip injections on postpartum day 3, the BSO-only group was given ip injections of BSO (4 mmol/kg body wt), the NACA-only group received ip injections of only NACA (250 mg/kg body wt), and the NACA + BSO group was given a dose of NACA 30 min before administration of the BSO injection. The pups were sacrificed on postpartum day 15, after examination under a slit-lamp microscope. Their lenses were analyzed for selective oxidative stress parameters, including glutathione (reduced and oxidized), protein carbonyls, catalase, glutathione peroxidase, glutathione reductase, and malondialdehyde. The lenses of pups in both the control and the NACA-only groups were clear, whereas all pups within the BSO-only group developed well-defined cataracts. It was found that supplemental NACA injections during BSO treatment prevented cataract formation in most of the rat pups in the NACA + BSO group. Only 20% of these pups developed cataracts, and the rest retained clear lenses. Further, GSH levels were significantly decreased in the BSO-only treated group, but rats that received NACA injections during BSO treatment had these levels of GSH replenished. Our findings indicate that NACA inhibits cataract formation by limiting protein carbonylation, lipid peroxidation, and redox system components, as well as replenishing antioxidant enzymes.

Glutathione depletion in vivo enhances contraction and attenuates endothelium-dependent relaxation of isolated rat aorta

Ten-day administration of the glutamate–cysteine ligase inhibitor l-buthionine–[S,R]-sulfoximine (BSO; 20 or 30 mM in drinking water) to adult male Sprague–Dawley rats induced 50–60% glutathione depletion ( p < 0.001) and elevated aortic ring reactive oxygen species release and tissue and plasma H 2O 2 concentrations ( p < 0.001) compared to control animals (CON) that consumed normal drinking water. In contrast to previous studies using tail cuff plethysmography methods, BSO had no significant effect on systolic blood pressure assessed by indwelling femoral artery catheters in conscious animals (10-day values, 119 ± 3 mn Hg vs 122 ± 4 mm Hg in CON vs BSO, respectively). Thoracic aorta rings were excised for in vitro assessment of vasomotor function. BSO shifted the phenylephrine (PE) dose–response curve to the left ( p = 0.003), lowering the EC 50 for PE contraction (from −6.752 ± 0.056 to −7.056 ± 0.055 log units; p = 0.001). Endothelium-dependent relaxation to acetylcholine (ACh) was significantly blunted ( p = 0.019) and the EC 50 for ACh relaxation was significantly increased (from −7.428 ± 0.117 to −7.129 ± 0.048 log units; p = 0.02) in BSO vs CON. Endothelium-independent vasorelaxation to sodium nitroprusside was similar in BSO and CON groups. Thoracic aorta immunoblot analyses revealed increases in endothelial nitric oxide synthase, superoxide dismutase 1 and 2, and soluble guanylate cyclase in BSO vs CON (all p < 0.01). Thus, enhanced PE contraction, blunted endothelium-dependent relaxation, and adaptations in nitric oxide bioavailability pathways provide the first evidence of chronic, in vivo BSO-induced, oxidative stress-mediated direct effects on the vasomotor function of arteries.

Effect of palm oil on oxidative stress-induced hypertension in sprague-dawley rats

Background Oxidative stress, associated with increased plasma isoprostane (ISO) and reductions in plasma glutathione (GSH), has been shown to cause severe hypertension in normal rats. Palm oil (PO), with an unsaturated-to-saturated fatty acid ratio close to one and rich in antioxidant vitamins, has been investigated for its beneficial effects on arterial thrombosis and atherosclerosis. In this study, the effect of PO on oxidative stress induced by inhibition of GSH synthesis (using buthionine sulfoximine [BSO]) was examined. Methods Sprague-Dawley rats were separated into two groups and received either natural vitamin-rich PO (Carotino, 5 g/kg daily) or water by gavage. After 4 weeks, they were further divided between receiving either BSO (30 mmol/L/day in the drinking water) or drug-free water for an additional week. Mean arterial pressure (MAP), heart rate (HR), and body weight (BW) were measured before and weekly during the experiment. The levels of plasma ISO, nitric oxide (NO), prostacyclin (PGI 2), and thromboxane A 2 (TXA 2) were determined by enzyme immunoassay, and plasma, heart, and kidney GSH by high-performance liquid chromatography. Results The PO reduced the age-dependent increase in MAP, and the pressor response to BSO, without changing the HR or BW compared to the BSO and control groups. It also elevated PGI 2, NO, and aortic cGMP, but decreased TXA 2 and aortic cAMP. In addition, the BSO-induced increase in ISO and TXA 2, and the reduction in kidney GSH were attenuated by PO. However, the PO effect on NO, PGI 2, cGMP, and TXA 2 was partly counteracted by BSO. Conclusions Palm oil reduces BSO-induced oxidative stress and attenuates hypertension by mechanisms involving changes in endothelium-derived factors.

The protective effect of glutathione administration on adriamycin-induced acute cardiac toxicity in rats

Adriamycin (ADR) is a potent antitumor antibiotic drug known to cause severe cardiac toxicity. Although ADR generates free radicals, the role of these radicals in the development of cardiac toxicity is still not well understood. In the present study, we evaluated the effect of glutathione (GSH) supplementation or depletion on ADR-induced cardiotoxicity in male Wistar rats. Cardiac toxicity was induced by a single intraperitoneal injection of ADR (20 mg kg−1) and manifested by an increase in heart rate, blood pressure elevation, and increased serum creatine kinase (CK) and lactate dehydrogenase (LDH). The extent of lipoprotein oxidation, lipid peroxide measured as malondialdhye (MDA), total homocysteine (tHcy), lipid profile, and atherogenic index were markedly elevated, whereas cardiac GSH content was dramatically decreased in ADR rats. Pre- and co-treatment of ADR rats with GSH (5 mm kg−1) (ADR +GSH) markedly reduced the levels of CK, LDH, lipoprotein oxidation susceptibility, cardiac MDA, tHcy and atherogenic index, and elevated GSH levels in cardiac tissues. In contrast, GSH depletion through administration of l-buthionine-(S,R)-sulfoximine (BSO) (15 mm kg−1) before and after ADR injection (ADR +BSO) greatly exacerbated ADR cardiotoxicity compared to the control and ADR groups. Finally, there were also severe cardiac histopathological changes in ADR and ADR +BSO groups, which were nearly restored by GSH treatment. These results suggest that GSH inhibits ADR cardiotoxicity and might serve as a novel combination with ADR to limit free radical-mediated organ injury.