Immunohistochemical expression of CD117 in borderline, low- and high-grade ovarian surface epithelial tumours: A clinicopathological study.

INTRODUCTION: Ovarian cancer is one of leading causes of cancer related death in gynecology. CD117 is a tyrosine kinase receptor that plays an important role in regulation of apoptosis, cell proliferation and adhesion by binding to its ligand-stem cell factor. Recent studies demonstrated its aberrant overexpression in various malignancies and concluded that it may play a pivotal role in carcinogenesis. AIM: To evaluate CD117 expression in ovarian surface epithelial tumours. MATERIALS AND METHODS: This retrospective study included 30 ovarian epithelial borderline, low and highly malignant tumours' formalin-fixed paraffin-blocks (FFPE) tissue blocks. Tissue sections were subjected to the routine haematoxylin-eosin stain and with the anti-CD117 immunohistochemically. RESULTS: There is a high significant difference in CD117 expression between borderline and malignant groups (P = 0.001). Additionally, there was significant difference in expression in relation to histopathological type (serous versus non-serous) in low-grade and the high-grade ovarian surface epithelial tumours (p=0.04, p=0.035 respectively). Tumour grade and stage strongly correlates with CD117 expression (p=0.014, p=0.019 respectively). CONCLUSION: We concluded that CD117 expression was significantly correlated with higher ovarian tumour grade and stage.

Spectral characterization and biological evaluation of biomolecules from the peels of three orange fruits: a comparative study.

The present work was designed to investigate the presence of bioactive chemicals in the reaction mixtures (RMs) of peels of Valencia, Mandarin, and African navel oranges, through GC-MS and FT-IR studies. Limonene, a unique compound, is present in the RMs of the three orange peels. Moreover, hexadecanoic acid 2-hydroxy-1-(hydroxymethyl) ethyl ester was identified in the RMs of all the three-orange peels. The RM of Mandarin orange exhibited potent cytotoxic effect against MCF-7 ATCC human breast cancer cells (HBC). All the three RMs exhibited moderate antibacterial activity against the human pathogenic bacteria Staphylococcus aureus (ATCC 25923), Staphylococcus epidermidis (ATCC 12228), Enterococcus faecalis (ATCC 29212), Escherichia coli (ATCC 25922), Klebsiella pneumoniae (ATCC 700603), Salmonella choleraesis (ATCC 10708), Pseudomonas aeruginosa (ATCC 27853), and Proteus mirabilis (ATCC 299).

Molecular and Seroprevalence of Toxoplasmosis in Goats' Blood and Milk in Iraq.

Toxoplasmosis is one of the most acute parasitic and zoonotic infections, which causes severe economic losses in animals due to abortion and reproductive problems worldwide. Therefore, this study was conducted in Baghdad province to detect the prevalence of Toxoplasmosis in blood and milk samples of 384 adult female goats using the serological indirect-enzyme-linked immunosorbent assay (iELISA) and the molecular polymerase chain reaction (PCR) test. The positive iELISA results were 20.57% in sera and 5.99% in milk samples. Regarding cross-classification results, the iELISA results revealed that 5.73% of goats were positive by testing both sera and milk samples, 14.32% and 0.26% were positive for testing sera and milk only, respectively, and 79.69% were negative by testing sera and milk. Targeting the B1 gene, the total positive results of the PCR assay showed that 13.92% and 30.43% of blood and milk samples, respectively, were positive at 546bp. Concerning cross-classification results, the total positive goats by testing of both sera and milk was 8.86%, while 5.06% of goats were positive only for testing of blood, and 86.08% were negative for testing of both samples. At the same time, no positive PCR results were detected in milk samples. In conclusion, there is a wide prevalence and incidence of Toxoplasmosis among goats in study areas. Furthermore, studies are essential to detect the parasite in different ages and sexes of goats and other domestic and wild animals using ELISA as a reliable, automated, and rapid test and PCR as a highly confirmative test.

Checkpoint kinase1 (CHK1) is an important biomarker in breast cancer having a role in chemotherapy response.

BACKGROUND: Checkpoint kinase1 (CHK1), which is a key component of DNA-damage-activated checkpoint signalling response, may have a role in breast cancer (BC) pathogenesis and influence response to chemotherapy. This study investigated the clinicopathological significance of phosphorylated CHK1 (pCHK1) protein in BC. METHOD: pCHK1 protein expression was assessed using immunohistochemistry in a large, well-characterized annotated series of early-stage primary operable invasive BC prepared as tissue microarray (n=1200). RESULT: pCHK1 showed nuclear and/or cytoplasmic expression. Tumours with nuclear expression showed positive associations with favourable prognostic features such as lower grade, lower mitotic activity, expression of hormone receptor and lack of expression of KI67 and PI3K (P<0.001). On the other hand, cytoplasmic expression was associated with features of poor prognosis such as higher grade, triple-negative phenotype and expression of KI67, p53, AKT and PI3K. pCHK1 expression showed an association with DNA damage response (ATM, RAD51, BRCA1, KU70/KU80, DNA-PKCα and BARD1) and sumoylation (UBC9 and PIASγ) biomarkers. Subcellular localisation of pCHK1 was associated with the expression of the nuclear transport protein KPNA2. Positive nuclear expression predicted better survival outcome in patients who did not receive chemotherapy in the whole series and in ER-positive tumours. In ER-negative and triple-negative subgroups, nuclear pCHK1 predicted shorter survival in patients who received cyclophosphamide, methotrexate and 5-florouracil chemotherapy. CONCLUSIONS: Our data suggest that pCHK1 may have prognostic and predictive significance in BC. Subcellular localisation of pCHK1 protein is related to its function.

Design and "in vivo" evaluation of two adapted catheters for intrauterine transcervical insemination in sheep.

In order to obtain better fertility, we evaluated two ovine artificial insemination (AI) catheters that were manufactured according to the anatomical structure of the ewe cervix. Morphometric data of the cervix in Churra and Assaf breeds were used to design two types of curved catheters: CAT06 with one curvature and ZIGZAG with five curvatures in a zigzag shape. Two commercial catheters (IMV(®) and Minitüb(®)) were used as controls. In experiment 1, cervical penetration and the degree of reflux were measured in a Cervical AI simulated assay both Churra (n=28) and Assaf ewes (n=28). In experiment 2, a fertility study was performed with three catheters (only one commercial control catheter - IMV) in 465 inseminations (Assaf); and a second study analyzed only the top two catheters (IMV and CAT06) in 428 inseminations (210 Assaf and 218 Churra). The ewes were synchronized using intravaginal sponges (40 FGA mg during 14 days) and 500 IU of eCG. Deeper penetration of the cervix was obtained with the new catheters compared with the commercial ones (1.5, 1.3, 3.5 and 3.2 cm for the IMV, Minitüb, CAT06 and ZIGZAG catheters, respectively). The cervical penetration and the reflux grade of each catheter showed no differences between breeds. In experiment 2, the degree of penetration had no correlation with fertility of different catheters. The best percentage of lambing ewes was obtained with the IMV and CAT06 catheters (39.5 and 48.1%, respectively. vs 27.2% for ZIGZAG catheter, in the Assaf breed). Regarding effect of breed, Assaf (39.3% and 49.5 for IMV and CAT06, respectively) showed better lambing rates than Churra (29.0% and 39.0%, respectively), and the CAT06 catheter showed significantly higher rates for each breed.

Rising bacterial resistance to common antibiotics in Al Ain, United Arab Emirates.

There is a dearth of local information in Al Amin, United Arab Emirates about antibiotic resistance patterns. In this retrospective study in a tertiary referral hospital, antibiotic susceptibility results were analysed over the 5-year period 2004-08 and compared with a previous study in the same hospital during 1999-2002. Staphylococcus aureus showed a significant decrease in sensitivity to oxacillin from 95.0% in the period 1999-2002 to 84.4% in 2008. Sensitivity of Acinetobacter spp. to imipenem dropped from 99.0% in 2004 to only 32.5% in 2008. During the same period, almost half of Escherichia coli isolates developed resistance to cefotoxime. Significant reductions in sensitivity to Pseudomonas aeruginosa between 1999 and 2008 were found for almost all the antibiotics tested. Klebsiella spp. did not show any significant change in resistance to any of the tested antibiotics. Serious efforts are needed to reduce the risk of the spread of resistant strains of bacteria.

Rising bacterial resistance to common antibiotics in Al Ain, United Arab Emirates

There is a dearth of local information in Al Ain, United Arab Emirates about antibiotic resistance patterns. In this retrospective study in a tertiary referral hospital, antibiotic susceptibility results were analysed over the 5-year period 2004-08 and compared with a previous study in the same hospital during 1999-2002. Staphylococcus aureus showed a significant decrease in sensitivity to oxacillin from 95.0% in the period 1999-2002 to 84.4% in 2008. Sensitivity of Acinetobacter spp. to imipenem dropped from 99.0% in 2004 to only 32.5% in 2008. During the same period, almost half of Escherichia coli isolates developed resistance to cefotoxime. Significant reductions in sensitivity to Pseudomonas aeruginosa between 1999 and 2008 were found for almost all the antibiotics tested. Klebsiella spp. did not show any significant change in resistance to any of the tested antibiotics. Serious efforts are needed to reduce the risk of the spread of resistant strains of bacteria

Influence of breed and age on morphometry and depth of inseminating catheter penetration in the ewe cervix: a postmortem study.

A detailed examination of the cervical canal in the ewe was carried out. This analysis could be used to design new catheters for artificial insemination (AI) to achieve deeper cervical penetration and therefore better fertility results. Three hundred and sixty-five cervices from four sheep breeds (Churra, Assaf, Merino, Castellana) obtained postmortem were used. Cervix morphometry and depth of cervical penetration using two types of catheters were determined. A conventional straight catheter for ovine artificial insemination (IMV), and a bent catheter, ending in a stainless steel needle, 9 cm in length and with an 8 mm tip bent 45 degrees , were used. The results showed that the morphometry of the cervix depends on breed and age of the ewe. The cervices of Churra breed were shorter and narrower, and had a higher number of folds than those of other breeds. Postmortem cervical penetration was deeper when the cervices were longer and wider, and with fewer folds (Merino and Castellana breeds). In ageing ewes, the cervix tended to become longer and wider, with loose folds. This decreased structural complexity and significantly improved cervical penetration. The bent catheter allowed significantly greater cervical penetration than the straight IMV one.

Parasite populations in the brown rat Rattus norvegicus from Doha, Qatar between years: the effect of host age, sex and density.

A total of 179 urban rats were sampled in the city of Doha in Qatar across the winter seasons (February-April) of 2002 and 2003. Only two parasites were identified, with overall prevalences of 35.8% and 41.3% for the cestode Hymenolepis diminuta and the flea Xenopsylla astia respectively. The prevalence of H. diminuta was markedly influenced by both year of study and host age, being higher in 2003 and amongst older rats. The abundance of infection of H. diminuta was influenced by the year of study, host age and sex. Worm burdens in adult rats were almost twice as heavy in males compared with females and adults of both sexes harboured heavier infections than juveniles. The prevalence of X. astia was influenced by both year and host age, being higher in juvenile rats in 2002 and in adults in 2003. The abundance of X. astia was significantly higher in 2003 and both male and female rats showed similar abundances, but in 2003 females were more heavily infested. Reasons for this are discussed in relation to the differing foraging strategies shown by male and female rats. The prevalence and abundance profiles for both H. diminuta and X. astia were higher overall in 2003 due to a significant increase in the rat population density, although this did not reflect in any increase in parasite species richness. Rats that were infected with H. diminuta were almost twice as likely to be infected with X. astia than those without the cestode, but when controlled for the effects of year, host age and sex, no quantitative interactions were detected between the two parasite species.

Factors influencing the success of vaginal and laparoscopic artificial insemination in churra ewes: a field assay.

Pregnancy rate following artificial insemination (AI) in sheep is variable depending on several factors. The Churra breed (milk breed of the North-West of Spain) yields lower fertility results compared with other local and European breeds (Manchega, Latxa, Merino, Lacaune, Sarde, etc.). In this work we studied the influence of many factors on the fertility of the Churra breed (insemination technique, year, farm, age, male, number of inseminations per ewe, lambing-insemination interval and technician), analyzing lambing data obtained after 44448 inseminations (39.67% cervical AI via vagina, AIV, and 60.33% intrauterine AI using laparoscopy, AIL) in a categorical model. The most important factors influencing fertility after AI were farm, year, season, AI technique, and technician. AIL showed significantly higher fertility results than AIV (44.89% versus 31.25%). Season significantly affected fertility in both cases, but differences were more evident in AIV. Fertility dropped 1.74% (AIV) and 2.07% (AIL) per year as the ewes aged. Finally, AI fertility decreased when the lambing-insemination interval was lower than 10 weeks.

Decay of sperm obtained from epididymes of wild ruminants depending on postmortem time.

We have carried out a study on the effect of postmortem time (PT) in some characteristics of epididymal sperm salvaged from hunted Iberian red deer and roe deer. Testis were collected, identified, refrigerated down to 5 degrees C, and sent to our laboratory by the wardens of the hunting reserves. This way, samples were delivered at different times postmortem. Sperm were extracted from the cauda epididymis by means of cuts. Analyzed parameters were: osmolality, pH, motility-both subjectively and with CASA, HOS test reactivity, acrosomal status and viability (assessed with propidium iodide). Osmolality and pH rose with prolonged postmortem time, possibly due to tissue decomposition. Most sperm quality parameters negatively correlated with PT. Besides, when comparing PT classes (groups of 24 h for red deer and 30 h for roe deer), we could appreciate that motility was more affected by PT than other quality variables. Progressive motility was especially impaired. We also classified the samples in high, medium and low quality for each PT group (considering progressive motility, intact acrosomes and reactivity to the HOS test), and it was clear that after 2 days the number of high quality samples was testimonial, and after several days, we almost found only low quality samples. In conclusion, epididymal sperm from Iberian red deer and roe deer undergo a decrease of quality with PT, but it could stay acceptable within many hours postmortem. There are implications for wildlife conservation programs, as epididymal sperm is a good source of germplasm. If valuable animals die and it is not possible to process their sperm immediately, it may still be possible to obtain viable spermatozoa many hours later.

Effect of the interval between estrus onset and artificial insemination on sex ratio and fertility in cattle: a field study.

We have carried out a field trial in cattle to study the effect of the interval between the onset of estrus and AI on sex ratio and fertility. Data were obtained from 716 cows that had been inseminated at different times between 8 and 44 h from the visual detection of estrus. Before analyzing the data, it was grouped in three intervals considering the time between estrus onset and AI (8-18, 18-30, and > or = 30 h). Our results show that the percentage of calved females (73.05%) is significantly superior for early inseminations (8-18 h), and it decreases 1.85% per hour from the onset of estrus. Delayed AIs (> or = 30 h) produce a significant deviation of the sex ratio towards the males (72.06%); nevertheless, fertility (percentage of successful pregnancies) diminishes significantly, from 66.19% (8-18 h) to 45.35% (> or = 30 h). In conclusion, variations in the interval between the onset of estrus and AI modify sex ratio. However, we must consider its effect on fertility.

Use of chromatin stability assay, mitochondrial stain JC-1, and fluorometric assessment of plasma membrane to evaluate frozen-thawed ram semen.

Cryopreservation of semen imposes deleterious effects on spermatozoa, either killing a certain proportion of cells or causing subtle damages on sperm function in the surviving population, changes not easily revealed by conventional assays. We have tested three functional assessment techniques in frozen-thawed ram semen from six adult rams, cryopreserved following eight different protocols (four extenders, and glycerol being added at two temperatures). Semen samples were thawed and the following analyses were carried out: motility (CASA), membrane integrity (Hoescht 33258 and fluorometry), chromatin status (chromatin stability test and fluorescence-assisted cell sorting, FACS) and mitochondrial activity (JC-1 and FACS). Fluorometry outcome did not correlate with the other parameters and showed large variation, albeit discriminating among cryopreservation techniques (P < 0.01). Mitochondrial activity correlated, but with low values, with total and progressive motility. However, good sperm motility and high velocity values were associated to high mitochondrial membrane potential. The chromatin stability assay was also successfully carried out, and had a good relationship with male factor (%COMP alpha(t) and SD alpha(t) parameters). In conclusion, fluorometric assessment of membrane integrity albeit rendering poor results, merits improvement, being a low-cost and handy technique, especially for work in the field. On the other hand, both assessments of chromatin stability and mitochondrial status (JC-1 staining), combined with FACS, are reliable techniques that can be used for the functional assessment of frozen-thawed ram semen.

Effect of epididymis handling conditions on the quality of ram spermatozoa recovered post-mortem.

Post-mortem spermatozoa recovery is an important technique for obtaining germplasm reserves from genetically valuable animals or endangered species. However, there are many factors that influence the outcome of this technique. We have studied the effect of the interval between animal's death and sperm recovery (0, 24 or 48 h) on the quality and freezability of ram spermatozoa from cauda epididymidis. Storage temperature of epididymis (room temperature or 5 degrees C) was also analysed. Spermatozoa were diluted with Tes-Tris-Fructose solution supplemented with egg yolk (10%) and glycerol (4%), and frozen using a programmable biofreezer (-20 degrees C/min). Pre-freeze and post-thaw sperm samples showed viable spermatozoa up to 48 h after the animal's death, although their quality declined significantly as post-mortem storage time increased. Epididymis sperm stored at 5 degrees C showed better motility and a lower percentage of abnormal forms than epididymis stored at room temperature after 24 and 48 h. The fertilizing ability of cauda epididymis ram spermatozoa obtained at 0 and 24h after the animal's death is similar to that of ejaculated spermatozoa. Therefore, a good protocol for post-mortem semen collection in rams when epididymal spermatozoa cannot be collected immediately, is to preserve the epididymis at 5 degrees C and process the samples in the first 24h after the animal's death.

Field and in vitro assay of three methods for freezing ram semen.

Glycerol has been the most widely used cryopreservation agent for spermatozoa and a wide range of factors affect its action on sperm viability and fertilizing capacity. We tested three methods for freezing ram semen packed in 0.25 ml straws (final cellular concentration: 100 x 10(6) spz/ml). Method M1: Two-thirds of the final volume of diluent was added as solution A (without glycerol) to the pure semen at 35 degrees C. The sample was cooled to 5 degrees C (-0.30 degrees C/min), one-third of final diluent volume was added as solution B (final concentration of glycerol 4%) and the sample was maintained at 5 degrees C for 2h. It was then frozen in a programmable biofreezer (-20 degrees C/min down to -100 degrees C). Method M2: The sample was diluted with a specific solution at 35 degrees C (final concentration of glycerol 3%), cooled to 5 degrees C (-0.20 degrees C/min) and left for 2h. After that, it was frozen in nitrogen vapours. Method M3: Semen was diluted 1:1 in a specific solution (concentration of glycerol 2%) and cooled to 5 degrees C (-0.25 degrees C/min). The sample was then diluted again in the same solution to the final cellular concentration (final concentration of glycerol 4%). It was left for 1h at 5 degrees C and then frozen in a programmable biofreezer (-20 degrees C/min down to -100 degrees C). Best total motility (TM) and progressive motility (PM) (75.8 and 55.18%) were obtained using Method M3. Methods M1 and M3 gave significantly higher values (P<0.05) for kinetic parameters: average path velocity (VAP) (81.3 and 85.2 microm/s), straight-line velocity (VSL) (72.8 and 77.3 microm/s) and linearity (LIN) (66.6 and 68.8%). Method M2 showed the lowest kinetic parameters of motility (VAP 74.4, VSL 67.3 and LIN 62.5) and the highest percentage of cells with damaged plasma membrane (53.8%). Method M1 gave the worst results in viability and acrosome status assessed using fluorescence probes (31.3%-dead cells with damaged acrosomes-versus 25.4% in M2 and 23.3% in M3). A field trial carried out on fertility showed a significantly higher percentage of pregnant or lambing ewes (P<0.05) with Method M3 (67.3% versus 51.1% for M1 and 58.8% for M2). We concluded that the use of a simple dilution medium (test-fructose-glycerol-egg yolk) with the addition of glycerol (to 2% at 35 degrees C and to 4% at 5 degrees C) in two steps together with a programmable biofreezer was a productive method for freezing ram semen.

Should doctors inform terminally ill patients? The opinions of nationals and doctors in the United Arab Emirates.

OBJECTIVES: To study the opinions of nationals (Emiratis) and doctors practising in the United Arab Emirates (UAE) with regard to informing terminally ill patients. DESIGN: Structured questionnaires administered during January 1995. SETTING: The UAE, a federation of small, rich, developing Arabian Gulf states. PARTICIPANTS: Convenience samples of 100 Emiratis (minimum age 15 years) and of 50 doctors practising in government hospitals and clinics. RESULTS: Doctors emerged as consistently less in favour of informing than the Emiratis were, whether the patient was described as almost certain to die during the next six months or as having a 50% chance of surviving, and even when it was specified that the patient was requesting information. In the latter situation, a third of doctors maintained that the patient should not be told. Increasing survival odds reduced the number of doctors selecting to inform; but it had no significant impact on Emiratis' choices. When Emiratis were asked whether they would personally want to be informed if they had only a short time to live, less than half responded in the way they had done to the in principle question. CONCLUSIONS: The doctors' responses are of concern because of the lack of reference to ethical principles or dilemmas, the disregard of patients' wishes and dependency on survival odds. The heterogeneity of Emiratis' responses calls into question the usefulness of invoking norms to explain inter-society differences. In the current study, people's in principle choices did not provide a useful guide to how they said they would personally wish to be treated.