Dysregulated Apoptosis Through the Intrinsic Pathway in the Internal Spermatic Vein of Patients With Varicocele

Background/PurposeApoptosis plays a critical role in various physiological processes. Varicocele is the most common cause of male infertility in adults. The dilated and thickened wall of the internal spermatic vein (ISV) in varicocele is considered similar to that in varicose veins. We investigated apoptotic protein expression in the ISV of varicocele, including Bcl-2, Fas, caspase-8 and caspase-9, to determine the intrinsic or extrinsic pathway.MethodsThe study group consisted of 10 patients with grade 3 left varicocele. The control group consisted of 10 patients with left indirect inguinal hernia. A 1-cm section of ISV was resected, using left inguinal incision, from each patient in both groups. The ISV sections were used to detect the mediators that regulate the intrinsic (Bcl-2 and caspase-9) and extrinsic (Fas and caspase-8) apoptotic pathways, by immunoblotting and immunohistochemical staining. Results were analyzed using Student's t tests.ResultsBcl-2, Fas, caspase-8 and caspase-9 immunoblots from both groups revealed a single band. The relative intensities of the Bcl-2 and caspase-9 protein bands differed significantly between the varicocele and control groups. Thickening of the smooth muscle layer of the ISV was found in patients with varicocele compared with the control group. Bcl-2 overexpression and downregulation of caspase-9 expression were noted in the varicocele group. There was no significant difference in Fas or caspase-8 expression in either group.ConclusionWe showed overexpression of Bcl-2 and downregulation of caspase-9 expression in the ISV under hypoxic stress. This indicated dysregulated apoptosis through the intrinsic pathway in the ISV of patients with varicocele. To the best of our knowledge, this is the first study of the apoptotic pathway in the human ISV. Additional studies are needed to establish whether adjunctive hyperbaric oxygen therapy reduces the recurrence rate after varicocelectomy

On the Infra-Red Spectra of Solutions of O-Chlorophenol and Phenol

<p><b>a–d Immunoblotting of different proteins in control and study groups of gastric adenocarcinoma.</b> (a) Representative immunoblots of different target proteins in gastric epithelium of the control and study groups (from patients 1 to 4) with GAPDH as a loading control. M, marker. Relative protein abundance of NKA α1 (b), NKA β1 (c), and E-cadherin (d) in gastric epithelium of the control and study groups. The asterisks indicate a significant difference between the control and study groups. Values were expressed as the means ± SEM. A.u., arbitrary unit. ***, P < 0.001; ****, P < 0.0001.</p

A Review of Metallothionein Isoforms and their Role in Pathophysiology

The Metallothionein (MT) is a protein which has several interesting biological effects and has been demonstrated increase focus on the role of MT in various biological systems in the past three decades. The studies on the role of MT were limited with few areas like apoptosis and antioxidants in selected organs even fifty years after its discovery. Now acknowledge the exploration of various isoforms of MT such as MT-I, MT-II, MT-III and MT-IV and other isoforms in various biological systems

Physiological Correlates of Volunteering

We review research on physiological correlates of volunteering, a neglected but promising research field. Some of these correlates seem to be causal factors influencing volunteering. Volunteers tend to have better physical health, both self-reported and expert-assessed, better mental health, and perform better on cognitive tasks. Research thus far has rarely examined neurological, neurochemical, hormonal, and genetic correlates of volunteering to any significant extent, especially controlling for other factors as potential confounds. Evolutionary theory and behavioral genetic research suggest the importance of such physiological factors in humans. Basically, many aspects of social relationships and social activities have effects on health (e.g., Newman and Roberts 2013; Uchino 2004), as the widely used biopsychosocial (BPS) model suggests (Institute of Medicine 2001). Studies of formal volunteering (FV), charitable giving, and altruistic behavior suggest that physiological characteristics are related to volunteering, including specific genes (such as oxytocin receptor [OXTR] genes, Arginine vasopressin receptor [AVPR] genes, dopamine D4 receptor [DRD4] genes, and 5-HTTLPR). We recommend that future research on physiological factors be extended to non-Western populations, focusing specifically on volunteering, and differentiating between different forms and types of volunteering and civic participation

The development and validation of a scoring tool to predict the operative duration of elective laparoscopic cholecystectomy

Background: The ability to accurately predict operative duration has the potential to optimise theatre efficiency and utilisation, thus reducing costs and increasing staff and patient satisfaction. With laparoscopic cholecystectomy being one of the most commonly performed procedures worldwide, a tool to predict operative duration could be extremely beneficial to healthcare organisations. Methods: Data collected from the CholeS study on patients undergoing cholecystectomy in UK and Irish hospitals between 04/2014 and 05/2014 were used to study operative duration. A multivariable binary logistic regression model was produced in order to identify significant independent predictors of long (> 90 min) operations. The resulting model was converted to a risk score, which was subsequently validated on second cohort of patients using ROC curves. Results: After exclusions, data were available for 7227 patients in the derivation (CholeS) cohort. The median operative duration was 60 min (interquartile range 45–85), with 17.7% of operations lasting longer than 90 min. Ten factors were found to be significant independent predictors of operative durations > 90 min, including ASA, age, previous surgical admissions, BMI, gallbladder wall thickness and CBD diameter. A risk score was then produced from these factors, and applied to a cohort of 2405 patients from a tertiary centre for external validation. This returned an area under the ROC curve of 0.708 (SE = 0.013, p  90 min increasing more than eightfold from 5.1 to 41.8% in the extremes of the score. Conclusion: The scoring tool produced in this study was found to be significantly predictive of long operative durations on validation in an external cohort. As such, the tool may have the potential to enable organisations to better organise theatre lists and deliver greater efficiencies in care

Surgical site infection after gastrointestinal surgery in high-income, middle-income, and low-income countries: a prospective, international, multicentre cohort study

Background: Surgical site infection (SSI) is one of the most common infections associated with health care, but its importance as a global health priority is not fully understood. We quantified the burden of SSI after gastrointestinal surgery in countries in all parts of the world. Methods: This international, prospective, multicentre cohort study included consecutive patients undergoing elective or emergency gastrointestinal resection within 2-week time periods at any health-care facility in any country. Countries with participating centres were stratified into high-income, middle-income, and low-income groups according to the UN's Human Development Index (HDI). Data variables from the GlobalSurg 1 study and other studies that have been found to affect the likelihood of SSI were entered into risk adjustment models. The primary outcome measure was the 30-day SSI incidence (defined by US Centers for Disease Control and Prevention criteria for superficial and deep incisional SSI). Relationships with explanatory variables were examined using Bayesian multilevel logistic regression models. This trial is registered with ClinicalTrials.gov, number NCT02662231. Findings: Between Jan 4, 2016, and July 31, 2016, 13 265 records were submitted for analysis. 12 539 patients from 343 hospitals in 66 countries were included. 7339 (58·5%) patient were from high-HDI countries (193 hospitals in 30 countries), 3918 (31·2%) patients were from middle-HDI countries (82 hospitals in 18 countries), and 1282 (10·2%) patients were from low-HDI countries (68 hospitals in 18 countries). In total, 1538 (12·3%) patients had SSI within 30 days of surgery. The incidence of SSI varied between countries with high (691 [9·4%] of 7339 patients), middle (549 [14·0%] of 3918 patients), and low (298 [23·2%] of 1282) HDI (p < 0·001). The highest SSI incidence in each HDI group was after dirty surgery (102 [17·8%] of 574 patients in high-HDI countries; 74 [31·4%] of 236 patients in middle-HDI countries; 72 [39·8%] of 181 patients in low-HDI countries). Following risk factor adjustment, patients in low-HDI countries were at greatest risk of SSI (adjusted odds ratio 1·60, 95% credible interval 1·05–2·37; p=0·030). 132 (21·6%) of 610 patients with an SSI and a microbiology culture result had an infection that was resistant to the prophylactic antibiotic used. Resistant infections were detected in 49 (16·6%) of 295 patients in high-HDI countries, in 37 (19·8%) of 187 patients in middle-HDI countries, and in 46 (35·9%) of 128 patients in low-HDI countries (p < 0·001). Interpretation: Countries with a low HDI carry a disproportionately greater burden of SSI than countries with a middle or high HDI and might have higher rates of antibiotic resistance. In view of WHO recommendations on SSI prevention that highlight the absence of high-quality interventional research, urgent, pragmatic, randomised trials based in LMICs are needed to assess measures aiming to reduce this preventable complication

Large expert-curated database for benchmarking document similarity detection in biomedical literature search

Document recommendation systems for locating relevant literature have mostly relied on methods developed a decade ago. This is largely due to the lack of a large offline gold-standard benchmark of relevant documents that cover a variety of research fields such that newly developed literature search techniques can be compared, improved and translated into practice. To overcome this bottleneck, we have established the RElevant LIterature SearcH consortium consisting of more than 1500 scientists from 84 countries, who have collectively annotated the relevance of over 180 000 PubMed-listed articles with regard to their respective seed (input) article/s. The majority of annotations were contributed by highly experienced, original authors of the seed articles. The collected data cover 76% of all unique PubMed Medical Subject Headings descriptors. No systematic biases were observed across different experience levels, research fields or time spent on annotations. More importantly, annotations of the same document pairs contributed by different scientists were highly concordant. We further show that the three representative baseline methods used to generate recommended articles for evaluation (Okapi Best Matching 25, Term Frequency-Inverse Document Frequency and PubMed Related Articles) had similar overall performances. Additionally, we found that these methods each tend to produce distinct collections of recommended articles, suggesting that a hybrid method may be required to completely capture all relevant articles. The established database server located at https://relishdb.ict.griffith.edu.au is freely available for the downloading of annotation data and the blind testing of new methods. We expect that this benchmark will be useful for stimulating the development of new powerful techniques for title and title/abstract-based search engines for relevant articles in biomedical research.Peer reviewe

Decreased expression of zonula occludens-1 and occludin in the bladder urothelium of patients with interstitial cystitis/painful bladder syndrome

Unique barrier properties of the urothelial surface membrane permit urine storage without contents leak into the bloodstream. Previous reports suggested that the bladder urothelial barrier might be compromised in interstitial cystitis/painful bladder syndrome (IC/PBS). We examined the changes of tight junction proteins (zonula occludens-1 (ZO-1) and occludin) in IC/PBS patients. Methods: Bladder samples were derived from of 32 patients with IC/PBS and eight controls. We detected the tight junction proteins of ZO-1 and occludin expression by immunoblotting, immunohistochemical (IHC) staining and double immunofluorescent (IF) staining with confocal microscopy. Data were analyzed using the Mann-Whitney U-test. Results: Expression of ZO-1 and occludin in the IC/PBS group was reduced compared to the control group by immunoblotting and IHC staining. Also, the thinning and denudation of urothelium were demonstrated in the IC/PBS group by histological study. IF staining showed the interruption of bladder urothelium in IC/PBS patients under confocal microscopy. Conclusion: Our data showed that decreased expression of tight junction proteins (ZO-1 and occludin) and interruption of bladder urothelium in IC/PBS patients. Treatment to repair the discontinuous urothelium may be useful to relieve some clinical symptoms of patients with IC/PBS

New insights to the mechanisms of varicocele-induced infertility

精索靜脈曲張(Varicocele)是指睪丸迴流的蔓狀靜脈叢血液受阻，造成內精索靜脈(internal spermatic vein；ISV)血管腫脹、彎曲及變長等現象。因解剖因素，90％疾病發生在左側。約有15~20％的年輕男性患有不同程度的精索靜脈曲張；而在不孕症的男性患者中更有30~40％同時患有精索靜脈曲張。精索靜脈曲張本身的手術治療並不困難，卻仍有2~14％不等的術後復發率；而對於這種常見的男性疾病引起不孕的相關機轉，至今也不甚明瞭。本論文的研究動機，即肇因於上述理由，希望經由探討內精索靜脈曲張的致病因素，以及睪丸組織凋亡(apoptosis)之路徑，更加瞭解精索靜脈曲張造成不孕的機轉。 過去的研究提出有關精索靜脈曲張損害精子生成的假說，包括睪丸溫度的增加、腎臟或腎上腺代謝物逆流、局部睪丸組織缺氧及性腺內分泌異常等。本論文以一系列實驗針對上述假說中的缺氧(hypoxia)因素，探討其影響機制。實驗包括(1)病人的內精索靜脈(ISV)之缺氧誘發因子-1αhypoxia-inducible factor-1α, HIF-1α的表現；以〝實驗誘導左側精索靜脈曲張〞experimental left varicocele; ELV)的老鼠，探討HIF- 1α在其ISV的表現。由於Kilinc等人(2004)及Barqawi等人(2004) 的研究；分別發現ELV老鼠其睪丸的HIF-1表現量與生殖細胞凋亡增加，顯示ELV老鼠的睪丸比正常老鼠的睪丸缺氧且造成生殖細胞凋亡增加，所以本論文接續探討(3)凋亡蛋白caspase-3在ELV老鼠睪丸缺氧環境下的表現；以及(4)ELV老鼠的睪丸組織細胞凋亡路徑(apoptotic pathway)。 由上述的四項實驗發現下列結果：(1) HIF-1α蛋白質表現量在精索靜脈曲張的患者之ISV較對照組高出七倍，主要分佈在血管的平滑肌層，表示這段發病的血管在缺氧的環境下，產生病理變化以適應環境；這也是首次探討有關人類ISV的研究；(2)利用ELV老鼠，發現HIF-1α在其ISV的表現量在術後第二、四及八週均顯著高於對照組，表示動物模式的ELV老鼠與病人相同，精索靜脈曲張的ISV均有缺氧的情形；由於ELV老鼠的睪丸HIF-1α表現量也比正常組高，因此推測男性患者睪丸的組織也發生缺氧的變化；(3)術後第八週及第十二週的ELV老鼠其睪丸的activated caspase-3表現量，顯著地高於對照組，配合TUNEL染色結果顯示生殖細胞的凋亡隨著老鼠精索靜脈曲張的時間而顯著增加。由ELV老鼠的實驗結果推論患有精索靜脈曲張的男性可能因生殖細胞凋亡比例增加而致不孕；(4)進一步分析ELV老鼠睪丸中不同凋亡路徑相關的蛋白質如Bcl-2，caspase-9，caspase-8及activated caspase-3的表現，發現隨著術後時間，實驗組織抗凋亡蛋白Bcl-2的蛋白質表現量下降，而caspase-9及activated caspase-3兩者蛋白質表現量上升；caspase-8在實驗組與對照組的表現量則無明顯差異，因此推測其細胞凋亡是經由內生性路徑(intrinsic apoptotic pathway)。 綜合以上的臨床及動物實驗結果，證實精索靜脈曲張造成靜脈及睪丸組織的缺氧，導致生殖細胞經由內生性的路徑凋亡增加。此一新解(new insights)也充分顯示精索靜脈曲張這種常見的男性疾病與不孕之間的關聯性。未來個人期望能以此研究成果為基礎，進一步朝這項疾病的治療和預防術後復發做出貢獻。Varicocele, described as an abnormal tortuosity and dilatation of internal spermatic veins (ISV) within the spermatic cord, is a common anomaly in adolescent and adult males, with an incidence of 15-20% of the general population. Several anatomic features contribute to the predominance of left-side varicoceles (more than 90%). It is the most common cause of male infertility in adults. Approximately 30%～40% of men evaluated for infertility have varicocele. The mechanisms that how the varicocele causing male infertility, however, are not clear. The surgical treatment for varicocele is safe and simple but the post-operative recurrence rates are 2%～14%. Based on the above-mentioned reasons, to investigate the pathogenesis of ISV and testicular cell apoptosis could led us to more understand more about the mechanisms of varicocele causing male infertility. Several theories of varicocele effects on the testicles have been postulated: elevated intratesticular temperature, gonadal hormonal imbalance, renal or adrenal vein blood refluxes to ISV, and local testis hypoxia. This paper includes a series of experiments that focus on the hypoxic factor in varicocele disease. These studies contain (1) expression of HIF-1α in the ISV of patients with varicocele; (2) expression of HIF-1α in the ISV of rats with experimental-induced left varicocele (ELV). Kilinc et al. (2004) and Barqawi et al. (2004) found that either HIF-1α expression or germ cell apoptosis increased in the testes of rats with ELV, respectively, indicating the occurrence of hypoxia and increase of cell apoptosis in the testes of rats with ELV. Hence, (3) activated caspase-3 (apoptotic protein) expression in testicular germ cell of rats with ELV; and (4) apoptotic pathway of testicular tissues in rats with ELV were studied. The results of the series of experiments are (1) HIF-1α expression in the ISV of patients with varicocele was significantly higher (7-fold) than in the control group. Moreover, expression of HIF-1α in the smooth muscle of the ISV was predominant in patients with varicocele. These data revealed that hypoxia related pathophysiological changes occurred in the ISV of patients with varicocele and that hypoxia might also occurr in the testicular tissue. To our knowledge, this is the first report on HIF-1α expression in human ISV; (2) expression of HIF-1α in the ISV of rats with ELV was significantly higher than that of the control group, especially at 2, 4, and 8 weeks after varicocele creation. This finding is compatible with the same hypoxic occurrence found in the ISV of patients with varicocele; (3) this animal study also demonstrated a significant increase on expression of activated caspase-3 protein in the testicular germ cells at 8 and 12 weeks after varicocele creation. Which indicated apoptosis of germ cells in the rats with ELV increased gradually. It may be a factor of varicocele causing male infertility in human due to increase apoptosis of testicular germ cell; (4) moreover, analyses of the apoptosis-related proteins expression in the testes of rats with ELV, e.g. Bcl-2, caspase-9, caspase-8, and activated caspase-3, showed down-regulation of Bcl-2 and gradually up-regulation of caspase-9 and activated caspase-3 expressions at 8 and 12 weeks after varicocele creation. There is no significant difference of caspase-8 expression between varicocele and control groups. So it is suggested that increased apoptosis of testicular tissue of rats with ELV was through the intrinsic pathway. According to the results of human and animal experiments, varicocele was demonstrated to lead to testicular tissue hypoxia and increase of germ cell apoptosis through intrinsic pathway. The new insights reveal the mechanisms of varicocele and the relationship between varicocele and male infertility. These findings will shad light on the treatment of varicocele and prevention of recurrence in the future.Chinese Abstract……....………………………………………………...….............i English Abstract…......….………………………………………………….…..…iii 1. Introduction………….………………………………………………...…...…..01 1.1Varicocele……………..…………… ………………………………….……01 1.2 Hypoxia.......………………………………........…….……………..…….…01 1.3 Apoptotic pathways.........................................................……….…….….…03 1.4 The aims of the study…………..........………………………………….….04 2. Materials and Methods……………………………….…………...…..….….…06 2.1 Patients and tissue samples..…………………………. ………………..….06 2.2 Technique and experiments of ELV rats...……………………………..….06 2.3 Immunoblotting ..…………………………………..…………………....….08 2.4 Immunohistochemistry…………………………….………………….…... 11 2.5 TUNEL assay..................................................................................................13 2.6 Statistical analyses.........................................................................................14 3. Results………………………………………………………………….……......15 3.1 Expression of HIF-1a in the internal spermatic vein of patients with varicocele………...………...........….........…………………………….…..15 3.2 Expression of HIF-1a in the internal spermatic vein of experimental left varicocele in rats…...…………..............................……………….............15 3.3 Activated caspase-3 expression in testicular germ cells of varicocele- induced rats..............................................................................................…16 3.4 Apoptotic pathway of testicular tissues in varicocele-induced rats..…....17 4. Discussion…………………….…….………………...…………………………18 4.1 Expression of HIF-1a in the internal spermatic vein of patients with varicocele..................................................................………………………18 4.2 Expression of HIF-1a in the internal spermatic vein of experimental left varicocele in rats….……………...................................……........……......20 4.3 Activated caspase-3 expression in testicular germ cells of varicocele- induced rats…..…..................…………….................……..………...…....21 4.4 Apoptotic pathway of testicular tissues in varicocele-induced rats..........23 5. Conclusions……...……………….………………………………………….…..26 6. References………………………...………………………………………......…27 7. Tables………………………..………….………………………………….….…37 8. Figures……...……………………………………………………………..……..4