Presence and role of stem cells in ovarian cancer

Ovarian cancer is the deadliest gynecological malignancy.It is typically diagnosed at advanced stages of the disease,with metastatic sites disseminated widely within the abdominal cavity.Ovarian cancer treatment is challenging due to high disease recurrence and further complicated pursuant to acquired chemoresistance.Cancer stem cell(CSC)theory proposes that both tumor development and progression are driven by undifferentiated stem cells capable of self-renewal and tumor-initiation.The most recent evidence revealed that CSCs in terms of ovarian cancer are not only responsible for primary tumor growth,metastasis and relapse of disease,but also for the development of chemoresistance.As the elimination of this cell population is critical for increasing treatment success,a deeper understanding of ovarian CSCs pathobiology,including epithelial-mesenchymal transition,signaling pathways and tumor microenvironment,is needed.Finally,before introducing new therapeutic agents for ovarian cancer,targeting CSCs,accurate identification of different ovarian stem cell subpopulations,including the very small embryoniclike stem cells suggested as progenitors,is necessary.To these ends,reliable markers of ovarian CSCs should be identified.In this review,we present the current knowledge and a critical discussion concerning ovarian CSCs and their clinical role.

Expression and localization of prostaglandin receptors and stromal factors in human cervix—Variations in pregnant and non-pregnant states

Prostaglandins (PGs) mediate cervical ripening leading to parturition. PGs are used successfully to induce cervical ripening. However, the cell type specific expression of PG receptor subtypes and various stromal factors important for cervical ripening in human cervix is not known. Our objective was to investigate the expression and localization of PG receptors EP1-4 and FP and localization of stromal factors CTGF (connective tissue growth factor), furin, calgranulin ?B, ALOX12 (arachidonate 12-lipooxy-genase) and ALOX15 in human cervical tissue from pregnant and non-pregnant women. Cervical biopsies were obtained from non-pregnant (NP), term pregnant (TP) and post-partum (PP) women. The mRNA expression was determined with real-time PCR, protein expression and localization with immunohistochemistry. Our results show that the EP2 mRNA level was higher in the PP group as compared to TP, whereas the EP4 mRNA level was lower in the TP group as compared to NP. Concomitantly stromal EP2 and epithelial EP3 immunoreactivity was higher in the TP as compared to the NP group, while the EP4 immunostaining in glands was lower in the TP as compared to the PP group. Immunostaining of endothelial CTGF, smooth muscle furin and ALOX12, were all lower in the TP group as compared to NP, for CTGF also the PP group was lower than NP. Endothelial calgranulin B immunoreactivity was higher in the PP group than the NP group. PG receptors and stromal factors exhibit differential expression in the cervix from women in non-pregnant and pregnant states, implying their involvement in the process of cervical ripening.

Improved cryopreservation of human hepatocytes using a new xeno free cryoprotectant solution

AIM:To optimize a xeno-free cryopreservation protocol for primary human hepatocytes.METHODS:The demand for cryopreserved hepatocytes is increasing for both clinical and research purposes.Despite several hepatocyte cryopreservation protocols being available,improvements are urgently needed.We first compared controlled rate freezing to polystyrene box freezing and did not find any significant change between the groups.Using the polystyrene box freezing,we compared two xeno-free freezing solutions for freezing of primary human hepatocytes:a new medium(STEM-CELLBANKER,CB),which contains dimethylsulphoxide(DMSO) and anhydrous dextrose,both permeating and non-permeating cryoprotectants,and the frequently used DMSO-University of Wisconsin(DMSOUW) medium.The viability of the hepatocytes was assessed by the trypan blue exclusion method as well as a calcein-esterase based live-dead assay before and after cryopreservation.The function of the hepatocytes was evaluated before and after cryopreservation by assessing enzymatic activity of 6 major cytochrome P450 isoforms(CYPs):CYP1A2,CYP2C9,CYP2C19,CYP2D6,CYP3A4 and CYP3A7.RESULTS:The new cryoprotectant combination preserved hepatocyte viability significantly better than the standard DMSO-UW protocol(P < 0.01).There was no significant difference in viability estimation between both the trypan blue(TB) and the Live-Dead Assay methods.There was a correlation between viability of fresh hepatocytes and the difference in cell viability between CB and DMSO protocols(r 2 = 0.69) using the TB method.However,due to high within-group variability in the activities of the major CYPs,any statistical between-group differences were precluded.Cryopreservation of human hepatocytes using the cryoprotectant combination was a simple and xeno-free procedure yielding better hepatocyte viability.Thus,it may be a better alternative to the standard DMSO-UW protocol.Estimating CYP activities did not seem to be a relevant way to compare hepatocyte function between different groups due to high normal variability between different liver samples.CONCLUSION:The cryoprotectant combination may be a better alternative to the standard DMSO-UW protocol in primary human hepatocyte cryopreservation.

Role of Surgery in the Elderly Patients Affected from Advanced Stage Ovarian Cancer

The aim of this study is to compare morbidity, surgical treatment and post-operative complications in elderly patients underwent surgery for advanced stage ovarian cancer, comparing to younger patients. Data of patients underwent surgery at the Department of Obstetrics and Gynecology of Cannizzaro Hospital (Catania) for advanced stage (IIIC-IV) ovarian cancer were collected from January 2000 to December 2013. Patients were stratified by age in two groups (I > 65 years and II < 65 years old). Following variables were collected: stage of the tumor, associated diseases, previous chemotherapy, the type of surgical procedures, blood transfusions, intraoperative and postoperative morbidity, mortality, and hospital stay. Median values between the two groups were compared using Mann-Whitney test and frequency data using?χ2. Statistical significance was defined as P < 0.05. A total of 179 patients were identified, they were divided into 2 groups: 64 patients were age 65 years or older (group I) and 115 patients were younger than age 65 (group II). In the whole series, 157 patients (87%) did not experience any complication. Overall, postoperative complications occurred in 10 (15%) patients in the group I and in 12 (10%) in the group II (p =?NS). In conclusion, elderly patients may tolerate well surgical procedures within acceptable postoperative morbidity, a length of hospital stay and a need for intensive care quite similar to that of younger patients.

Female urinary incontinence during pregnancy and after delivery: Clinical impact and contributing factors

Urinary incontinence(UI) is a common condition affecting adult women of all ages and it could have a negative infl uence on quality of life. The etiology of UI is multifactorial, but some of the most important risk factors are obesity and ageing, as well as adverse obstetric events. Pregnancy and delivery per se have been implicated in the etiology of UI. Although several studies have demonstrated a direct association between UI and vaginal delivery in short, medium and long-term, the role of childbirth on the risk of UI remains controversial. The mechanical strain during delivery may induce injuries to the muscle, connective and neural structures. Vaginal birth can be associated with relaxation or disruption of fascial and ligamentous supports of pelvic organs. Parity, instrumental delivery, prolonged labor and increased birth weights have always been considered risk factors for pelvic floor injury. Also genetic factors have been recently raised up but still there are not appropriate guidelines or measures to reduce signifi cantly the incidence of UI. The role of pelvic fl oor muscle training(PFMT) in the prevention and treatment of UI is still unclear. However, PFMT seems to be useful when supervised training is conducted and it could be incorporated as a routine part of women's exercise programmes during pregnancy and after childbirth.

Survival of HIV Infected Children Born to Mothers Enrolled in a PMTCT Program in a Resource Poor Setting

Background: Pediatric HIV is a leading cause of morbidity and mortality worldwide. The substantial expansion in PMTCT has generated information on rates of transmission and associated factors, but there are limited studies on disease progression and mortality in vertically infected children, especially from resource poor settings. Methods: A birth cohort study was initiated in 2002 to focus on the role of a single dose of nevirapine in HIV transmission before Highly Active Antiretroviral Therapy (HAART) was readily available. The enrolment of women and subsequent follow up of the children occurred at 3 peri urban clinics around Harare. Findings: 479 women were HIV infected. From these, 93 (19%) children became HIV infected, 182 (38.0%) uninfected and 204 (43%) lost to follow up before HIV diagnosis. Of the HIV infected children, 40 (43%) died before the fifth birthday, 26 (28%) were lost to follow up and 27 (29%) were alive five years after maternal enrolment prior to availability of cART. Conclusion: In this setting, there was unacceptable high mortality from HIV infected children and loss to follow up prior to availability of HAART. A small proportion of HIV vertically infected children is surviving in resource poor settings without antiretroviral therapy.

Twin gestation: Is induction of labor possible in highly selected cases?

Induction of labor has been in practice for several decades but the decision to induce labor in multifetal gestations has not been accepted by many practitioners in contemporary obstetrics. The aim of this study was to compare maternal and fetal outcomes among women with uncomplicated twin gestations who were induced at term and those who were not. It was a cross-sectional analytic study that lasted ten years, (1st January 2000 to 31st December 2009 inclusive) at the maternity of the central hospital, Yaounde, Cameroon. 158 women were enrolled, 79 were induced and 79 went into labor spontaneously. Maternal and fetal outcomes, duration of labor, the prevalence of caesarean section were compared in the two groups. The average age of the women in the groups was 26, 75 ± 3.65 years (range 15 - 41 years). For those who were induced, indication was premature rupture of membranes in 44 (56%) of cases. The characteristics of induced and none induced women were similar, except for the number of antenatal consultations (P ≤ 0.001). There was a significant difference in the duration of labor in the two groups (6 hrs versus 9, 75 hrs;P ≤ 0.001). The overall rate of vaginal delivery in the two groups was 87, 97% (n = 139), 88.87% in the group that was induced versus 86.07% in the group that labor was spontaneous (OR 1, 22;95% IC 0, 51-2, 92;p ≤ 0.90). With respect to primary outcomes, there were no statistically significant differences between the group that labor was induced and that which labor was spontaneous. The prevalence of caesarean delivery was similar in the two groups (10%, 12% vs 13%, 92%;OR 0%, 7%;95% IC 0, 27-1, 85;P ≤ 0.50). The main indications for caesarean section were acute fetal distress and failed induction. The first and fifth minute APGAR scores less than 7 in the first twins delivered in the two groups were similar. The 312 children delivered in both arms did not show any complications at birth. However, there were 3 neonatal deaths (1 vs 2). There were no cases of uterine rupture or maternal deaths among the 156 women. Induction of labor may be proposed to women with uncomplicated term twin gestations with specific indications. Induction in these highly selected cases does not impute any additional risks but close monitoring of labor is very necessary.

Cervical cancer screening:A never-ending developing program

With the term "oncological screening", we define the overall performances made to detect early onset of tumors. These tests are conducted on a population that does not have any signs or symptoms related to a neoplasm. The whole population above a certain age, only one sex, only subjects with a high risk of developing cancer due to genetic, professional, discretionary reasons may be involved. Screening campaigns should be associated, when risk factors that can be avoided are known, with campaigns for the prevention of cancer by means of suitable behavior. The goal of cancer screening cannot however be limited to the diagnosis of a greater number of neoplasms. Screening will be useful only if it leads to a reduction in overall mortality or at least in mortality related to the tumor. Screening should then allow the diagnosis of the disease at a stage when there is a possibility of healing, possibility that is instead difficult when the disease is diagnosed at the appearance of signs or symptoms. This is the reason why not all campaigns of cancer screening have the same effectiveness. In Italy, every year there are about 150000 deaths due to cancer. Some of these tumors can be cured with a very high percentage of success if diagnosed in time. Cervical cancer can be diagnosed with non-invasive tests. The screening test used all over the world is Papanicolaou(Pap) test. This test may be carried out over the entire healthy population potentially exposed to the risk of contracting cancer. Public health has begun the screening campaigns in the hope of saving many of the approximately 270000 new cases of cancer reported each year. Screening is done following protocols that guarantee quality at the national level: these protocols are subject to change over time to reflect new realities or to correct any errors in the system. A simplified sketch of a possible route of cancer screening is as follows:(1) after selecting the target population, for example all women between 25 and 64 years(in the case of monitoring of cervical cancer), an invitation letter with the date and time of the appointment, planned according to the acceptance capacity of the hospital, is sent to all individuals;(2) an examination, which depending on the individual and the type of cancer to be monitored, for example, canbe a Pap smear, is performed and the patient can go home;(3) once available the results of examinations, if negative, they shall be communicated to the person concerned that will be notified by mail and will be recalled for a second test at a few years of distance, in the case of non-negativity, instead, the patient is contacted by telephone and informed of the need to carry out further examinations: it is said that the patient is in the "phase two" of the screening pathway;(4) in phase two, reached by only a small portion of the interested parties(usually less than 3%-5%), more indepth tests are carried out, which, depending on the individual and the type of cancer, can be: cytological and colposcopic examinations, the removal of a fragment of tissue(biopsy) and subsequent histological examination, additional tests such as ultrasound, radiography, or others such as computerized tomography, magnetic resonance imaging, positron emission tomography, etc., in case of negativity, the concerned person will be called for new control tests at a a few years of distance, in case of non-negativity, it will be proposed instead an oncologic therapeutic plan and/or surgery to treat the diagnosed tumor; and(5) once the treatment plan is completed, the individual enters the follow-up protocol, which is monitored over time to see if the tumor has been completely removed or if instead it is still developing. Cervical cancer is undoubtedly the most successful example of a cancer screening campaign. Paradoxically, its effectiveness is one of the strongest reasons to criticize the usefulness of vaccination against human papillomavirus(HPV) in countries where the screening service with Pap test is organized in an efficient manner. Cervical cancer screening protocols are directed to sexually active women aged 25-64 years: they provide the Pap test performed by examining under a microscope or by staining with a specific "thin prep" the material taken from the cervix with a small spatula and a brush. It is recommended to repeat the test every two or three years. It is important to emphasize that women vaccinated against HPV MUST continue the screening with Pap test. Although some screening programs(e.g., Pap smears) have had remarkable success in reducing mortality from a specific cancer, any kind of screening is free from inherent limitations. The screening methods are in fact applied to large parts of the apparently healthy population. In particular, the limits for certain cancers may be as obvious as to prohibit the introduction of an organized screening program. Potential limitations of organized screenings are basically of two types: organizational and medical. The limits of organizational type relate to the ability of a program to recruit the whole target population. Although well organized, a screening program will hardly be able to exceed a coverage of 70%-80% of the target population, and in fact the results of the current programs are often much smaller. The limits of medical type are represented by the possibility of reducing the overall mortality, or specific mortality, using a specific screening campaign.

Chip-based multimodal super-resolution microscopy for histological investigations of cryopreserved tissue sections

Histology involves the observation of structural features in tissues using a microscope.While diffraction-limited optical microscopes are commonly used in histological investigations,their resolving capabilities are insufficient to visualize details at subcellular level.Although a novel set of super-resolution optical microscopy techniques can fulfill the resolution demands in such cases,the system complexity,high operating cost,lack of multi-modality,and low-throughput imaging of these methods limit their wide adoption for histological analysis.In this study,we introduce the photonic chip as a feasible high-throughput microscopy platform for super-resolution imaging of histological samples.Using cryopreserved ultrathin tissue sections of human placenta,mouse kidney,pig heart,and zebrafish eye retina prepared by the Tokuyasu method,we demonstrate diverse imaging capabilities of the photonic chip including total internal reflection fluorescence microscopy,intensity fluctuation-based optical nanoscopy,single-molecule localization microscopy,and correlative light-electron microscopy.Our results validate the photonic chip as a feasible imaging platform for tissue sections and pave the way for the adoption of super-resolution high-throughput multimodal analysis of cryopreserved tissue samples both in research and clinical settings.

Nat Biotechnol:科学家成功绘制出单细胞中短链RNA分子的图谱

近日，刊登在国际杂志Nature Biotechnology上的一项研究报告中，来自瑞典卡罗琳学院的研究人员通过研究成功在单个胚胎干细胞中测定了短链非编码RNA序列的绝对数量。当基因中的信息被使用时，比如当其编码蛋白质时，首先DNA会转录成为信使RNA来作为蛋白质制造的范本，