The importance of oxidative stress in patients with chronic renal failure whose hypertension is treated with peritoneal dialysis

Increased oxidative stress is a well-known phenomenon in dialysis patients. However, the contribution of hypertension to the oxidative stress in peritoneal dialysis patients has not yet been assessed. The present study aimed to investigate if hypertension had an additional effect on oxidative stress in peritoneal dialysis patients. A total of 50 patients treated with peritoneal dialysis were divided into two groups: The patients with mean of last three blood pressure results as 135/90 mmHg and above were considered hypertensive, the patients with lower blood pressure were considered normotensive. The control group included 25 healthy individuals. Serum malondialdehyde (MDA), advanced oxidation protein product (AOPP), myeloperoxidase (MPO), catalase (CAT) and glutathione peroxidase (GSH-Px) levels were measured in all groups. MDA level, an indicator of lipid peroxidation, was significantly higher in the hypertensive group compared to the control group, while the increase in the normotensive group was not significant. However, the difference between the hypertensive and normotensive groups was significant. The levels of AOPP, an indicator of protein oxidation level, and MPO, an indicator of neutrophil activation, were not different between the groups, while the activities of antioxidant CAT and GSH-Px decreased in both normotensive and hypertensive groups compared to the control group, and there was no significant difference between the patient groups. This study shows that both normotensive and hypertensive peritoneal dialysis patients have increased-oxidative stress and decreased antioxidant levels and hypertension might have an additional effect on oxidative stress by increasing MDA level in peritoneal dialysis patients.     

Proteomic analysis in peritoneal dialysis patients with different peritoneal transport characteristics

Peritoneal membranes can be categorized as high, high average, low average, and low transporters, based on the removal or transport rate of solutes. In this study, we used proteomic analysis to determine the differences in proteins removed by different types of peritoneal membranes. Peritoneal transport characteristics in patients who received peritoneal dialysis therapy were assessed by a peritoneal equilibration test. Two-dimensional differential gel electrophoresis technology followed by quantitative analysis was performed to study the variation in protein expression from peritoneal dialysis effluents (PDE) among different groups. Proteins were identified by MALDI-TOF-MS/MS analyses. Further validation in PDE or serum was performed utilizing ELISA analysis. Proteomics analysis revealed ten protein spots with significant differences in intensity levels among different groups, including vitamin D-binding protein, complement C3, apolipoprotein-A1, complement factor C4A, haptoglobin, alpha-1 antitrypsin, immunoglobulin kappa light chain, alpha-2-microglobulin, retinol-binding protein 4 and transthyretin. The levels of vitamin D-binding protein, complement C3, and apolipoprotein-A1 in PDE derived from different groups were greatly varied (P < 0.05). However, no significant difference was found in the serum levels of these proteins among different groups (P > 0.05 for all groups). This study provides a novel overview of the differences in PDE proteomes of four types of peritoneal membranes. Vitamin D-binding protein, complement C3, and apolipoprotein-A1 showed enhanced expression in PDE of patients with high transporter.     

Phospholipids profiling and outcome of peritoneal dialysis patients

Abstract

Objectives: To investigate phospholipids (PLs) biomarkers in predicting outcome of patients undergoing peritoneal dialysis (PD).

Materials and methods: Twenty PD patients were followed using baseline plasma PLs with an improved online two-dimensional liquid chromatography-quadrupole time-of-flight mass spectrometry system.

Results: Significant differences were observed in eight PL species with sphingomyelin (SM) and glycerophosphocholine between technical survival (n=15) and failure patients (n?=?5). Cox regression showed SM 21:0 (adjusted HR 13.7, 95% CI 2.42–77.88, p?=?0.003) was independently associated with patients technical failure.

Conclusions: PD failure patients had different plasma PLs profiling as compared with survival patients. Elevated plasma SM 21:0 level may potentially serve as a biomarker of PD patients at risk for adverse outcomes.     

维持性腹膜透析患者并发真菌性腹膜炎的易感因素和结局分析

目的分析腹膜透析相关性真菌性腹膜炎(FP)发生率、致病菌、治疗情况和预后。方法回顾性分析2010年1月至2019年10月陆军军医大学第二附属医院腹膜透析中心发生的18例FP,选择与同期收治非真菌性腹膜炎113例比较,记录所有FP患者的临床资料,治疗方法和转归,分析FP发生的易感因素和结局。结果腹膜透析相关性腹膜炎共389例次,FP 18例次,占4.6%。其中白念珠菌6例(33.3%)、近平滑念珠菌5例(27.8%)、无名念珠菌3例(16.7%)、光滑念珠菌2例(11.1%)、热带念珠菌1例(5.6%)和克柔念珠菌(5.6%)1例。与非真菌性腹膜炎相比较,FP组腹透时间更长(P<0.001)、既往抗生素使用率高(P<0.001)、血浆白蛋白(ALB)更低(P<0.001)、C反应蛋白(CRP)更高(P<0.001)、甲状旁腺激素(PTH)和血磷(P)水平更高(P<0.001)。Logistic回归分析结果显示腹透时间越长、1个月内使用抗生素、低ALB和高CRP是发生FP的危险因素(P<0.05)。18例次FP中,14例患者拔管转血透(77.8%),4例患者死亡(22.2%),FP组腹膜透析技术失败率和死亡率明显高于BP组。结论腹透时间越长、既往使用抗生素、低ALB和高CRP是FP的易感因素。FP是腹膜透析的严重并发症,是导致技术失败的主要原因,确诊后早期拔管可降低死亡率。     

Update on the challenging role of biofilms in peritoneal dialysis

Biofilms are commonly associated with an increased risk of patient infection. In peritoneal dialysis (PD), catheter associated infection, especially peritonitis, remains a clinically relevant problem. Although the presence of a biofilm is recognized in relapsing, repeat, and catheter-related peritonitis, it remains poorly characterized. In this review, an update on the role of biofilms in PD infections is presented. The emerging concept that host cells and tissue associated biofilms, in addition to the biofilms on the catheters themselves, contribute to the recalcitrance of infections is discussed. Furthermore, the evidence of biofilms on PD catheters, their developmental stages, and the possible influence of the PD environment are reviewed. The focus is given to ex vivo and in vitro studies that contribute to the elucidation of the interplay between host, microbial, and dialysis factors. The key issues that are still to be answered and the challenges to clinical practice are discussed.     

miR-15a-5p suppresses inflammation and fibrosis of peritoneal mesothelial cells induced by peritoneal dialysis via targeting VEGFA

Long-term peritoneal dialysis (PD) often ends up with ultrafiltration failure (UFF) which is partially caused by persistent inflammation and fibrosis of peritoneal tissues. However, the mechanism is still unclear. In the current study, the peritoneum from UFF patients demonstrated inflammation and fibrosis which were positively related to the expression of vascular endothelial growth factor A (VEGFA). The in vitro model using human peritoneal mesothelial cells (HPMCs) stimulated by high glucose or advanced glycation end (AGE) product showed consistent changes of inflammation, fibrosis, and VEGFA. What's more, we showed that VEGFA was an instigator of inflammation and fibrosis. Several microRNAs (miRNAs) have been reported to regulate expression of VEGFA elsewhere. Five of them were selected to test the expression in the peritoneum of patients with PD. Results suggested that miR-15a-5p was the most significantly downregulated one. Also, in high glucose or AGE product-stimulated HPMCs, miR-15a-5p decreased. When miRNA mimic was used to restore the expression of miR-15a-5p, high glucose-induced VEGFA was repressed. The predicted binding site between these two molecules was confirmed by the dual-luciferase assay. Restoration of miR-15a-5p restrained inflammation and fibrosis of HPMCs. TGF-β1/Smad2 was shown to be the downstream signaling pathway and their activity was regulated by miR-15a-5p/VEGFA. In conclusion, our current study demonstrates that miR-15a-5p acts as a regulator of VEGFA mRNA and the following inflammation and fibrosis in peritoneal mesothelial cells. The miR-15a-5p/VEGFA pathway may be a potential target for preventing ultrafiltration failure in patients with PD.     

Speciation of chromium in plasma and liver tissue of endstage renal failure patients on continuous ambulatory peritoneal dialysis

This work is a first attempt to determine the speciation of Cr in human plasma. With the aid of in vitro and in vivo⁵¹Cr-labeled experiments, it was possible to develop the necessary biochemical techniques for the separation of the plasma proteins. Further work will use real samples, taking care to avoid contamination of the various fractions and to preserve the original binding of the Cr to the specific plasma compounds. In a first attempt on the distribution of Cr over the different organelles of liver tissue, work will be restricted to in vivo labeled experiments with rats. The procedure to do the speciation work seems so elaborate that it may be impossible ever to achieve the contemplated speciation of Cr in human liver tissue by subcellular fractionation.     

Restoration of the cellular thiol status of peritoneal macrophages from CAPD patients by the flavonoids silibinin and silymarin

During continuous ambulatory peritoneal dialysis (CAPD) the peritoneal immune cells, mainly macrophages, are highly compromised by multiple factors including oxidative stress, resulting in a loss of functional activity. One reason for the increase of inflammatory reactions could be an imbalance in the thiol-disulfide status. Here, the possible protective effects of the antioxidant flavonoid complex silymarin and its major component silibinin on the cellular thiol status were investigated. Peritoneal macrophages from dialysis fluid of 30 CAPD patients were treated with silymarin or silibinin up to 35 days.

A time-dependent increase of intracellular thiols was observed with a nearly linear increment up to 2.5-fold after 96 hours, reaching a maximum of 3.5-fold after 20 days of culture. Surface-located thiols were also elevated. The stabilization of the cellular thiol status was followed by an improvement of phagocytosis and the degree of maturation as well as significant changes in the synthesis of IL-6 and IL-1ra. Furthermore, the treatment of peritoneal macrophages with flavonoids in combination with cysteine donors resulted in a shortened and more efficient time course of thiol normalization as well as in a further increased phagocytosis. In addition, GSH-depletion in thiol-deficient media simulating CAPD procedures led to intracellular thiol deficiency similar to the in vivo situation.

It is concluded that treatment with milk thistle extracts silymarin and silibinin alone or, more effectively in combination with cysteine donors, provide a benefit for peritoneal macrophages of CAPD-patients due to a normalization and activation of the cellular thiol status followed by a restoration of specific functional capabilities.     

念珠菌血症的危险因素和预后分析

目的了解念珠菌血症的临床特点、分布及预后危险因素。方法回顾性调查2012年1月至2014年5月浙江大学医学院附属第一医院所有血培养念珠菌阳性的患者资料,分析其临床特征、治疗和预后等,采用χ2检验或Fisher精确概率法进行预后单因素分析,采用多元Logistic回归进行预后多因素分析。结果 97例念珠菌血症患者入选,其中男性64例,女性33例,平均年龄（59.6±16.8）岁。包括白色念珠菌51例（52.6%）,非白色念珠菌46例（47.4%）,非白色念珠菌中热带念珠菌17例（17.5%）、近平滑念珠菌12例（12.4%）、光滑念珠菌7例（7.2%）、无名念珠菌4例（4.1%）、其他念珠菌6例（6.2%）。念珠菌培养阳性后30 d内死亡37例,30 d病死率为38.1%。Logistic多因素回归分析显示：年龄（OR=1.104,95%CI：1.041~1.170,P=0.001）、血液系统肿瘤（OR=63.256,95%CI：2.898~1380.833,P=0.008）、APACHEⅡ评分（OR=1.176,95%CI：1.053~1.313,P=0.004）、感染性休克（OR=12.032,95%CI：2.389~60.587,P=0.003）及合并细菌性血流感染（OR=26.016,95%CI：4.002~169.127,P=0.001）是其死亡的独立危险因素;而拔除或更换深静脉置管（OR=0.118,95%CI：0.025~0.559,P=0.007）是念珠菌血症死亡的独立保护性因素。结论念珠菌血症患者分布科室范围广、基础疾病重、侵入性操作多。年龄、高APACHEⅡ评分、感染性休克及合并细菌血流感染是影响念珠菌血症死亡的独立危险因素,拔除或更换深静脉置管是念珠菌血症死亡的独立保护性因素。     

骨化三醇联合腹膜透析疗法治疗慢性肾功能衰竭的临床疗效及对患者血清ProGRP、Cysc、Chemerin水平的影响

目的:探讨骨化三醇联合腹膜透析疗法治疗慢性肾功能衰竭的临床疗效及对患者血清Pro-Gastrin-Releasing Peptide (ProGRP)、CystatinC(Cysc)、Chemerin水平的影响。方法:选取我院2017年2月至2018年1月收治的98例慢性肾功能衰竭患者,按照随机数表法将其分为观察组(n=51)和对照组(n=47)。对照组采用腹膜透析疗法治疗,观察组采用骨化三醇联合腹膜透析疗法治疗。观察和比较两组治疗前后肾功能指标尿素氮(blood urea nitrogen,BUN)、血清肌酐(Serum creatinine Cr)、24小时尿蛋白(24 h urinary protein,24 h UP),生化指标白蛋白(albumin,Alb)、血红蛋白(hemoglobin,Hb)及红细胞(red blood cell,RBC),胃泌素释放肽前体(ProGRP)、血清胱抑素(Cys C)、趋化素(Chemerin)水平的变化,6个月、1年生存率及不良反应的发生情况。结果:治疗后,观察组BUN、SCr、24hUP水平均显著低于对照组[(13.95±3.06)mmol/L vs.(21.10±3.85)mmol/L,(260.12±40.32)μmol/L vs.(354.93±51.06)μmol/L,(1.75±0.45)g/24 h vs.(2.67±0.80)g/24 h](P0.05);Alb水平显著低于对照组[(27.85±3.58)g/L vs.(33.06±4.27)g/L](P0.05);Hb、RBC显著高于对照组[(91.72±13.46)g/L vs.(82.36±10.15)g/L,(379.47±92.08)×1012/L vs.(315.70±73.24)×1012/L](P0.05);ProGRP、Chemerin水平显著低于对照组[(49.23±4.72)pg/mL vs.(63.87±7.30)pg/mL,(37.02±6.15)μg/L vs.(30.63±4.81)μg/L](P0.05);Cysc水平显著高于对照组[(80.75±16.08)mL/min vs.(98.81±18.07)mL/min](P0.05);6个月、1年生存率均显著高于对照组[96.08%(49/51) vs. 91.49%(43/47),90.20%(46/51) vs. 74.47%(35/47)](P0.05);不良反应总发生率显著低于对照组[17.65%(9/51) vs. 44.68%(21/47)](P0.05)。结论:骨化三醇联合腹膜透析疗法治疗慢性肾功能衰竭的临床效果显著优于单用,其可有效减轻患者的临床症状,纠正电解质紊乱,改善肾功能和预后,可能与降低血清ProGRP、Chemerin水平及提高血清Cysc水平有关。     

Aberrant T cell activation and heightened apoptotic turnover in end-stage renal failure patients: a comparative evaluation between non-dialysis,haemodialysis, and peritoneal dialysis

Patients in end-stage renal disease (ESRD) have a high incidence of bacterial and viral infections. Fifteen non-dialysed (ND), 15 haemodialysed (HD), 15 patients with peritoneal dialysis (PD), and 15 healthy controls were included. T cell proliferation was measured by [3H]thymidine uptake. Apoptosis and cell phenotype were determined by FACS. sTNF-R1, sCD95, interleukin-1beta-converting enzyme (sICE), and interleukin (IL)-10 were measured by ELISA. PHA and CD3-driven T cell proliferation were significantly decreased in ESRD patients. CD3(+), CD19(+) B cells, and percentage of CD4(+) T cells were significantly reduced. Percent memory T cells (CD45RO(+)) and cells undergoing apoptosis (CD95(+)/Annexin V+) were significantly increased in ESRF. Moreover, sCD95, sTNFRI, and ICE were significantly increased. Serum level of IL-10, a Th2 cytokine, was enhanced. These findings strongly suggest that in ESRD patients Th1 T cells are selectively susceptible to undergo apoptosis. This observation provides an additional pathophysiological concept in the genesis of Th2 dominance.     

Preliminary observations on the association between serum IL-6 and hydration status and cardiovascular risk in patients treated with peritoneal dialysis

IntroductionSystemic inflammation, as defined by elevated blood IL-6, is a strong independent predictor of peritoneal dialysis (PD) patient survival. The present study has aimed to determine whether there exists a particular “phenotype” associated with high systemic IL-6 that characterizes PD patients in terms of their fluid status and cardiac parameters.MethodsFifty-seven prevalent PD patients were classified according to serum concentrations of IL-6. The degree of overhydration was assessed by bioimpedance analysis (BIA). Echocardiography and serum concentrations of NT-proBNP and troponin T were used to assess cardiovascular risk.ResultsPatients with high serum IL-6 were older, more often diabetic, treated with PD for longer, and significantly more overhydrated. There was a significant correlation between serum IL-6, hydration status (r = 0.38; p = 0.002) and serum albumin (r = −0.35; p = 0.009). Multivariate regression analysis confirmed a strong association of overhydration, hypoalbuminemia, and systemic IL-6 concentration. Patients with high IL-6 had significantly increased levels of both NT-proBNP (r = 0.36; p = 0.006) and TnT (r = 0.50; p < 0.001) in the absence of abnormalities in echocardiography.ConclusionsHigh systemic IL-6 identifies PD patients with increased cardiovascular risk that is significantly related to overhydration. Thus, the measurement of serum IL-6 may contribute to the more accurate assessment of cardiovascular status in patients undergoing PD.     

老年人下肢动脉病变预后随访及危险因素分析

目的:分析老年人下肢动脉痛变发生的影响因素及其预后的影响.方法:回顾性分析2004～2009年于哈尔滨医科大学第一临床医学院干部二病房明确诊断为下肢动脉病变的住院患者181例.全部行双下肢动脉彩色多普勒检查,按照动脉病变程度将患者分为3组:斑块组、狭窄组(≥50%)和闭塞组.收集其性别、年龄、血脂、空腹血糖(FBG)、纤维蛋白原(FIB)、高血压、脑卒中及吸烟史等信息.比较不同病变组一般情况及生化指标间的差异,并对老年下肢动脉粥样硬化病变危险因素采用回归方法进行分析.所有患者均接受16个月随访,观察患者的主要终点事件包括下肢坏疽、侧支循环、死亡.结果:①年龄、糖尿病病程、总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL-C)、FBG、血脂异常、性别比例、糖尿病、冠心病、脑卒中合并率在三组间的差异无统计学意义(P>0.05).②三组间高血压痛程、高血压合并率、吸烟时间、吸烟率、HDL-C、FIB在三组之间的差异有统计学意义(P<0.05).③Logistic回归分析结果显示TG、高血压、纤维蛋白原与老年LEAD的发生成正相关(P<0.05),HDL-C与LEAD成负相关(P<0.05).④经过16个月随访,三组间生存率无显著差异(P>0.05).结论:老年LEAD患者男性明显多于女性,且常有较高的吸烟率、高血压、糖尿病、脑卒中、血脂异常的合并率.TG、FIB水平增高及合并高血压痛其下肢血管病变程度更重,是加剧下肢动脉硬化的主要因素,HDL-C时LEAD具有保护作用.经过16个月随访,老年LEAD患者死亡的主要原因为心脑血管疾病,与血管狭窄严重程度无相关性.     

The frequency of Th2 type cells increases with time on peritoneal dialysis in patients with diabetic nephropathy.

We have analysed the frequency of cytokine-producing T cells in different dialysis groups (haemodialysis; HD and peritoneal dialysis; PD) over time. Although we saw no difference in type 1 cytokine production (IL-2 and IFN-gamma) in either dialysis group, there was a clear increase in the percentage of T cells spontaneously producing the type 02 cytokines in the PD group (IL-4, r = 0.558, P < 0.05; IL-10, r = 0.527, p < 0.05). Our patient group was carefully selected to include patients with an ongoing autoimmune disease, insulin dependent diabetes mellitus (IDDM) (DN group) and chronic glomerulonephritis (GN), which are common reasons of end stage renal failure. As expected there was no increase in the spontaneous production of either IL-4 or IL-10 in either disease group with patients undergoing HD treatment. However, there was a clear correlation with the frequency of T cells producing IL-4 (r = 0.755, P < 0.05) and IL-10 (r = 0.725, P < 0.05) and time on dialysis in the PD patients with DN, but not those with GN. Much work has suggested that the pathogenesis of IDDM is associated with a Th1 dominated response. We show here that this response is skewed towards a Th2 response after long term treatment with PD. This work demonstrates that the immunological effects of different dialysis modalities on patients with different diseases vary. This may go some way to explain why certain patient groups have more complications with different dialysis modalities.     

The role of WNT5A and Ror2 in peritoneal membrane injury

Patients on peritoneal dialysis are at risk of developing peritoneal fibrosis and angiogenesis, which can lead to dysfunction of the peritoneal membrane. Recent evidence has identified cross-talk between transforming growth factor beta (TGFB) and the WNT/β-catenin pathway to induce fibrosis and angiogenesis. Limited evidence exists describing the role of non-canonical WNT signalling in peritoneal membrane injury. Non-canonical WNT5A is suggested to have different effects depending on the receptor environment. WNT5A has been implicated in antagonizing canonical WNT/β-catenin signalling in the presence of receptor tyrosine kinase-like orphan receptor (Ror2). We co-expressed TGFB and WNT5A using adenovirus and examined its role in the development of peritoneal fibrosis and angiogenesis. Treatment of mouse peritoneum with AdWNT5A decreased the submesothelial thickening and angiogenesis induced by AdTGFB. WNT5A appeared to block WNT/β-catenin signalling by inhibiting phosphorylation of glycogen synthase kinase 3 beta (GSK3B) and reducing levels of total β-catenin and target proteins. To examine the function of Ror2, we silenced Ror2 in a human mesothelial cell line. We treated cells with AdWNT5A and observed a significant increase in fibronectin compared with AdWNT5A alone. We also analysed fibronectin and vascular endothelial growth factor (VEGF) in a TGFB model of mesothelial cell injury. Both fibronectin and VEGF were significantly increased in response to Ror2 silencing when cells were exposed to TGFB. Our results suggest that WNT5A inhibits peritoneal injury and this is associated with a decrease in WNT/β-catenin signalling. In human mesothelial cells, Ror2 is involved in regulating levels of fibronectin and VEGF.     

16S rDNA高通量测序分析腹膜透析胃肠功能障碍患者肠道菌群特征

采用16S rDNA高通量测序技术分析腹膜透析（PD）胃肠功能障碍患者的肠道菌群变化特征,探讨肠道菌群在腹膜透析胃肠功能障碍中的具体作用。

收集25例PD胃肠功能正常者（PDGF组）、25例PD胃肠功能障碍者（PDGD组）和13例健康者（Normal组）的粪便样本,提取肠道菌群基因组,应用16S rDNA高通量测序技术对各组测序结果进行菌群多样性、物种组成差异和菌群功能分析。

3组的Observed species指数（H = 6.905,P = 0.032）和Shannon指数（H = 6.993,P = 0.030）差异具有统计学意义。与Normal组相比,PDGD组Shannon指数显著降低（P = 0.027）,PDGF组Observed species指数显著升高（P = 0.044）;与PDGF组相比,PDGD组Observed species和Shannon指数显著降低（P＜0.05）。PCoA结果显示3组各自聚集,区分较明显。UPGMA聚类分析结果显示3组大部分样本均在本组中聚类后再与其他组进一步合并。LEfSe分析结果显示,Normal组优势菌群包括颤螺菌目（Oscillospirales）、瘤胃菌科（Ruminococcaceae）、栖粪杆菌属（Faecalibacterium）、拟杆菌科（Bacteroidaceae）等;PDGF组优势菌群包括毛螺菌目（Lachnospirales）、毛螺菌科（Lachnospiraceae）、布劳特菌属（Blautia）;PDGD组优势菌群包括芽孢杆菌纲（Bacilli）、乳杆菌目（Lactobacillales）、链球菌属（Streptococcus）、链球菌科（Streptococcaceae）等。KEGG L1水平的功能组成集中在代谢、遗传信息处理和环境信息处理;KEGG L2水平的功能主要集中在碳水化合物、氨基酸代谢、遗传信息的复制和修复及转运、膜运输途径;与Normal组和PDGF组相比,PDGD组在碳水化合物、氨基酸代谢途径富集减少,感染性疾病、聚糖生物合成和代谢途径富集增加。

PD胃肠功能障碍患者存在肠道菌群失调,调整肠道微生态是防治PD患者出现胃肠功能障碍的潜在干预靶标。

     

Apoptosis and cell proliferation correlated with tumour grade in peritoneal fluids of patients with serous ovarian cancer

A. Kalogeraki, I. Karvela‐Kalogeraki, P. E. Petraki, I. Zois, D. Tamiolakis and E. N. Stathopoulos
Apoptosis and cell proliferation correlated with tumour grade in peritoneal fluids of patients with serous ovarian cancer Objective: Apoptosis and cell proliferation in peritoneal fluids of patients with ovarian serous adenocarcinoma have not been well described in cytology. To investigate the contribution of cell death to the growth of this tumour we analysed both apoptosis and cell proliferation in peritoneal fluids of patients with ovarian serous adenocarcinoma. Methods: We studied 40 tumours from 40 patients with ovarian serous adenocarcinoma. Twelve tumours were high grade, 13 were moderately differentiated and 15 were poorly differentiated. The detection of DNA fragments in situ using the terminal deoxyribonucleotidy transferase (TDT)‐mediated dUTP‐digoxigenin nick‐end labelling (TUNEL) assay was applied to investigate active cell death (apoptosis), and the MIB‐1 antigen was used to investigate cell proliferation. Results: The TUNEL indices were 0.29 ± 0.05, 0.79 ± 0.10 and 2.1 ± 0.90 in Grade I, Grade II and Grade III ovary carcinomas, respectively. The MIB‐1 antigen labelling indices were 6.5 ± 0.09, 12.9 ± 3 and 25.8 ± 6.2, respectively, in the same order of tumour differentiation. The differences in both TUNEL and MIB‐1 labelling indices were statistically significant between Grade I, Grade II and Grade III carcinomas and there was a positive correlation between the two indices (P < 0.001). Conclusions: Apoptosis and cell proliferation increased as the grade of tumour increased in ovarian serous adenocarcinoma, suggesting a rapid turnover of the tumour cells in tumours of higher grade, and may play an important role in the growth and the extension of such cancer cells in the peritoneal cavity.     

Screening for Fabry disease in patients undergoing dialysis for chronic renal failure in Turkey: Identification of new case with novel mutation

Background

Chronic renal failure (CRF) is a serious complication of Fabry disease (FD). The aims of the present study were to determine the prevalence of unrecognized FD in Turkish hemodialysis population and to investigate the molecular background.

Method

Primarily, α-galactosidase A (α-Gal A) activity was investigated on DBS in 1136 patients of both sexes who underwent dialysis for CRF in Turkey. The disease was confirmed by analyzing enzyme activity in leukocyte and GLA gene sequencing in all patients in whom α-Gal A level was 40% of normal or less.

Results

Mean age of the patients (44.5% female, 52.5% male) was 56.46 ± 15.85 years. Enzyme activity was found low with DBS method in 12 patients (four males, eight females). Two men, but no women, were diagnosed with FD by enzymatic and molecular analysis. In consequence of genetic analysis of a case, a new mutation [hemizygote c.638C>T (p.P214S) missense mutation in exon 5] was identified, which was not described in literature. Family screening of cases identified six additional cases.

Conclusion

As a result of this initial screening study performed on hemodialysis patients for the first time with DBS method in Turkey, the prevalence of FD was detected as 0.17%. Although the prevalence seems to be low, screening studies are of great importance for detecting hidden cases as well as for identifying other effected family members.     

Prevalence of, and risk factors associated with, perinatal calf mortality in pasture-based Holstein-Friesian cows

Recent publications indicate that the prevalence of perinatal mortality has increased in some dairy industries and an increased proportion of this loss is not associated with the traditional risk factors for perinatal mortality. The objectives of this study were to establish the prevalence of perinatal mortality (calf death within 24 h of calving) in Irish dairy herds and to determine the current significance of putative risk factors in pasture-based management systems. A total of 182 026 records of full-term calvings from Holstein-Friesian dams served by artificial insemination (AI) sires of seven breeds in herds of 20 calvings or more per year were available from the Irish national breeding database over 4 years (2002 to 2005). The prevalence of perinatal mortality was 4.29% (7.7% in primiparae and 3.5% in pluriparae). The likelihood of perinatal mortality increased between 2002 and 2005 and was greatest in June and in winter. There was an interaction (P < 0.001) between the effect of calving assistance and parity with the effect of dystocia on perinatal mortality being greater in primiparae. The odds of perinatal mortality were greater in male (OR = 1.12; P < 0.001) and in twin calves (OR = 5.70-13.36; P < 0.001) and in dams that had perinatal mortality at the previous calving (OR = 4.21; P < 0.001). The logit of the probability of perinatal mortality increased by 0.099 per unit increase in sire predicted transmitting ability (PTA) for direct perinatal mortality. The probability of perinatal mortality increased at an increasing rate in primiparae as animals calved at a younger age relative to the median age at first calving. The only herd-level factor examined, herd size did not affect the odds of perinatal mortality. These data indicate that the prevalence of perinatal mortality in this cattle population is similar to that in other pasture-based dairy systems worldwide. The putative exposures and attributes traditionally associated with perinatal mortality were associated with perinatal mortality in this pasture-based dairy cow population. The practical implication of these results is that as many of the significant risk factors are largely not under management control (year of calving, month of calving, twin calving, primiparity, previous perinatal mortality and foetal gender), herd owners must focus on the significant determinants under their control (age at first calving, sire genetic merit for direct perinatal mortality and both the extent of calving supervision and the degree of assistance), in order to reduce the prevalence of perinatal mortality and improve perinatal welfare.