Mutations at the mitochondrial DNA polymerase (POLG) locus associated with male infertility.

Human mitochondrial DNA polymerase, encoded by POLG, contains a polyglutamine tract encoded by a CAG microsatellite repeat. Analysis of POLG genotypes in different populations identified an association between absence of the common, ten-repeat allele and male infertility typified by a range of sperm quality defects but excluding azoospermia.

Assisted reproductive technology treatment outcomes.

Information on the outcomes of ART treatments in Ireland is not readily available to Irish practitioners. The data for hospital affiliated clinics has been made available for many years and is included in the hospital reports. We present a 10-year analysis of the Irish ART results voluntarily reported by six out of seven IVF clinics. The data was collected from published ESHRE reports and from results (2007-8) not yet published. Data collected included: number of clinics and ART cycles, female age, clinical and multiple pregnancy rates and treatment complications. The clinical pregnancy rate per embryo transfer was 31.7% for IVF and 29.8% for ICSI. The proportion of singleton, twin and triplet deliveries for IVF and ICSI combined was 75%, 23.35% and 1.64%. The rate of ovarian hyperstimulation was 0.8%. ART practice in Ireland is safe, effective and responsible. Financial and societal savings could result from the introduction of state funded IVF with compulsory eSET where recommended.

Free triiodothyronine has a distinct circadian rhythm that is delayed but parallels thyrotropin levels.

CONTEXT: TSH is known to have a circadian rhythm, but the relationship between this and any rhythm in T(4) and T(3) has not been clearly demonstrated. OBJECTIVE: With a view to optimizing thyroid hormone replacement therapy, we have used modern assays for free T(4) (FT4) and free T(3) (FT3) to investigate circadian rhythmicity. SETTING: The study was performed at a university hospital. DESIGN AND SUBJECTS: This was a cross-sectional study in 33 healthy individuals with 24-h blood sampling (TSH in 33 and FT4 and FT3 in 29 individuals) and cosinor analysis. RESULTS: Of the individuals, 100% showed a sinusoidal signal in TSH, for FT4 76%, and for FT3 86% (P < 0.05). For FT4 and FT3, the amplitude was low. For TSH the acrophase occurred at a clock time of 0240 h, and for FT3 approximately 90 minutes later at 0404 h. The group cosinor model predicts that TSH hormone levels remain above the mesor between 2020 and 0820 h, and for FT3 from 2200-1000 h. Cross correlation of FT3 with TSH showed that the peak correlation occurred with a delay of 0.5-2.5 h. When time-adjusted profiles of TSH and FT3 were compared, there was a strong correlation between FT3 and TSH levels (rho = 0.80; P < 0.0001). In contrast, cross correlation revealed no temporal relationship between FT4 and TSH. CONCLUSIONS: FT3 shows a circadian rhythm with a periodicity that lags behind TSH, suggesting that the periodic rhythm of FT3 is due to the proportion of T(3) derived from the thyroid. Optimizing thyroid hormone replacement may need to take these rhythms into account.

Frozen-thawed transfer cycles: are they comparable with fresh?

Cryopreservation of zygotes and subsequent thaw and transfer is an established ART treatment. We assessed if success rates frozen-thawed (day 2) zygotes are comparable with the outcome in fresh cycles of treatment. We performed a prospective follow-up and analysis of all frozen (FZT) and fresh cycles of treatment during a 12 months period. One hundred and nineteen patients in the frozen-thawed and 652 in the fresh group had a transfer. The overall thaw-survival rate was 71.7%. Clinical pregnancy rates per thaw and transfer were respectively 15.1% and 21% in the frozen and 29.1% (per transfer) in the fresh group. Implantation rates in fresh and frozen cycles were 16% and 12.3% respectively. The pregnancy loss rate was higher in the FZT group (29% vs. 18.3%). Cryopreservation of good quality zygotes, after fresh transfer offers optimal success rates in subsequent frozen treatment. It also encourages consideration of elective single zygote transfers.

From egg to offspring. Survival rates in in-vitro fertilisation: where does it go wrong?

This retrospective study examines the attrition that occurs from all eggs inseminated and fertilised through to babies taken home in an IVF fresh/frozen programme undertaken between 2001-4. Poor survival rates are to be found, particularly during the recognised in-vitro developmental stages, Present practice which promotes quantity from the start, increases problem potential, is wasteful and against the concept of one embryo one baby. This suggests the need to develop better criteria for egg selection for insemination thus enabling a rethink to the approach to ovarian stimulation of one of quality rather than quantity.

Maternal folate status and neural tube defects in Ireland: the need for a national food fortification program.

To determine the present risk of a Neural Tube Defect [NTD] pregnancy in the caucasian primigravid population in Dublin by comparing the serum folate and red cell folate status of primigravid patients attending the first prenatal booking clinic with data from the late 80's. This Cross-sectional population study looking at blood folate status of over 400 sequential primigravid caucasian women with a singleton pregnancy, booking at less than or equal to 20 weeks gestation. All patients were attending a prenatal booking clinic at the Rotunda Hospital in Dublin during 2003-2004. Comparing serum and red cell folate values in 454 primigravid patients in 2003-4 to values to in a large case-control study based on over 56,000 women attending maternity hospitals in Dublin from 1986 to 1990. Just 13.9% of our patients took periconceptual folate, 33.5% of patients took folate in the first 20 weeks of pregnancy and 58.8% of mothers were taking no folate supplement. Overall, 30% of mothers had RCF levels below 400 ug/L--a level recommended as the minimum value required for protection. NTD risk occurred most frequently amongst patients with RCF levels between 300 and 400 ng/mL.

Using the NANA toolkit at home to predict older adults' future depression.

BACKGROUND: Depression is currently underdiagnosed among older adults. As part of the Novel Assessment of Nutrition and Aging (NANA) validation study, 40 older adults self-reported their mood using a touchscreen computer over three, one-week periods. Here, we demonstrate the potential of these data to predict future depression status. METHODS: We analysed data from the NANA validation study using a machine learning approach. We applied the least absolute shrinkage and selection operator with a logistic model to averages of six measures of mood, with depression status according to the Geriatric Depression Scale 10 weeks later as the outcome variable. We tested multiple values of the selection parameter in order to produce a model with low deviance. We used a cross-validation framework to avoid overspecialisation, and receiver operating characteristic (ROC) curve analysis to determine the quality of the fitted model. RESULTS: The model we report contained coefficients for two variables: sadness and tiredness, as well as a constant. The cross-validated area under the ROC curve for this model was 0.88 (CI: 0.69-0.97). LIMITATIONS: While results are based on a small sample, the methodology for the selection of variables appears suitable for the problem at hand, suggesting promise for a wider study and ultimate deployment with older adults at increased risk of depression. CONCLUSIONS: We have identified self-reported scales of sadness and tiredness as sensitive measures which have the potential to predict future depression status in older adults, partially addressing the problem of underdiagnosis.

Expression of receptor activator of nuclear factor kappabeta ligand (RANKL) and tumour necrosis factor related, apoptosis inducing ligand (TRAIL) in breast cancer, and their relations with osteoprotegerin, oestrogen receptor, and clinicopathological variables.

BACKGROUND: Receptor activator of nuclear factor kappabeta ligand (RANKL) has an important role in bone remodelling, and tumour necrosis factor related, apoptosis inducing ligand (TRAIL) can induce apoptosis in cancer cells. Their functions are linked by their interactions with osteoprotegerin (OPG). OBJECTIVE: To investigate the expression of RANKL and TRAIL in a large series of unselected breast cancers and to analyse the relations between these expressions and the expression of OPG, oestrogen receptor, and clinicopathological variables. METHODS: 395 breast cancers were sampled into tissue microarrays and immunohistochemistry undertaken for RANKL and TRAIL. RESULTS: There was strong expression of RANKL in 14% of the cancers and strong expression of TRAIL in 30%. Expression of RANKL had a negative association with expression of oestrogen receptor (p = 0.036). Expression of TRAIL had a negative association with the Nottingham Prognostic Index (p = 0.021). There was a significant negative relation between expression of RANKL and TRAIL (p<0.005). Unsupervised cluster analysis produced a dendrogram that showed a clear division into two groups, and the expression of oestrogen receptor was significantly higher in one of those groups (p = 0.012). CONCLUSIONS: There is apparent loss of expression of RANKL in 86% of breast cancers; those tumours that retain expression tend to be oestrogen receptor negative and of a high histological grade. There is strong expression of TRAIL in 30% of breast cancers and these tend to be of better prognostic type. These results may be important in the processes of metastasis to bone and the apoptotic cell death pathway in cancer.

The hidden costs of IVF.

Whether the payment is personal or funded via a third party, IVF is expensive. Clinical and drug costs are known and if necessary can be budgeted for. But, indirect expenses involved in participation also need to be considered. In order to find out what these latter might be, women attending Human Assisted Reproduction Ireland at the Rotunda Hospital Dublin completed prospectively a detailed questionnaire on all likely areas of extra expense during a single IVF cycle from down-regulation to zygote transfer. Four groups participated, divided according to their distance from the clinic. Group A were within Dublin and Group B within a 50 mile radius. Group C were between a 50-130 mile radius and Group D a 130-200 mile radius. Mean levels of costs increased with distance:- Group A 104 euros. to Group D 703 euros. Food, meals and accommodation were the main contributors. Travel time and time-off work also increased with distance. These were respectively, Group A to Group D 15hrs to 139hrs: 35hrs to 75hrs. The study demonstrates that significant extra expense in time and money can be spent in an IVF cycle over and above medical and drug costs. This must be advised prior to commencement. All other issues being equal, geographical location should be part of the decision as to which clinic to attend.

Clinical uses of estrogens.

PIP: Estrogens, whether natural or synthetic, have a wide range of clinical uses in the human female. In prepubertal females, estrogens are used in treating gonadal dysgenesis, excessive height, and genital infections. During the reproductive years, estrogens are used in managing 1) menstrual disorders (amenorrhea, menorrhagia, dysmenorrhea); 2) infertility (poor cervical mucus and anovulation); 3) pregnancy (abortion, lactation suppression); 4) dermatological disorders (acne vulgaris, hirsutism); 5) combined estrogen/progestogen usage for contraception; and 6) postcoital contraception. During the climacteric/postmenopausal years, estrogens are used in treating menopausal syndrome and breast cancer as well as various genital problems (infection, atropic vaginitis, genital prolapse). In the human male, estrogens are used in treating prostatic carcinoma and sexual problems. Estrogen therapy should be used with caution, and benefits should be weighed against the hazards. Possible side effects and alternative forms of treatment should be considered.^ieng

Preservation of fertility in women affected by cancer. An audit of the cryopreservation services provided for women at Human Assisted Reproduction Ireland.

There has been a remarkable improvement in survival rates for adult and childhood cancers, due to progress in cancer treatments. However multiagent chemotherapy and radiotherapy are associated with gonadal failure. Whereas sperm banking is commonly performed, female gametes are not so amenable to cryopreservation. Options available to women for fertility preservation include, embryo and oocyte cryopreservation. Our unit established an oocyte/embryo cryopreservation service for women affected by cancer in October 2003. Ninteen women have been referred and five (26%) women have had oocytes cryopreserved. The mean age of women in the group cryopreserved was 34 years (range 21-41). Their mean day 3 FSH was 6.2 IU (range 5.5-6.8). The mean dose of rFSH used was 1870 units (range 1500-2600). The mean number of oocytes cryopreserved was 8 (range 3-16). Preservation of fertility is of major concern to women affected by cancer. This service offers them hope for their future fertility.

The role of dynamic and morphological studies in the assessment of potential liver donors.

In a prospective study, 66 donor livers were evaluated by monoethylglycinexylidide (MEGX) dynamic clearance and semiquantitative scoring of pathological changes in liver biopsies. The median MEGX level in 63 donors was 89 mcg/L (range 16-250 mcg/L); fifteen had MEGX levels < 50 mcg/L, 17 between 50 and 90 mcg/L, and 31 > 90 mcg/L. There were no cases of primary nonfunction, and no deaths were related to poor graft function. There was no statistically significant difference in peak aspartate aminotransferase (AST), day 5 AST, peak bilirubin, or lowest prothrombin time among the 3 groups. Liver biopsies were assessed in 61 donors: 33 (54%) were normal and 17 (28%) showed mild, 8 (13%) showed moderate, and 3 (5%) showed severe steatosis. Postperfusion biopsy assessing the extent of preservation injury was essentially normal or showed minimal change in 16 (26%), mild change in 29 (48%), moderate in 13 (21%) and severe abnormalities in 3 (5%). The latter 3 biopsies all had severe steatosis. There was no significant difference in early graft function or outcome between moderate/severe groups and normal/minimal groups, although the former had a higher peak AST (P < 0.02) and peak bilirubin (P < 0.004). This detailed prospective analysis suggests that MEGX and the morphological studies may assist in the assessment of potential liver donors but they do not provide a basis on which grafts should be discarded.

Retinal hemorrhage thresholds for Q-switched neodymium-Yag laser exposures.

Thresholds for retinal vitreal and contained hemorrhages were determined for 1064 nm laser light at 30-nsec and 4-nsec pulsewidths. Rhesus monkeys received graded exposures from a neodymium-yag laser onto either the macular or extramacular region of the retina. Contained hemorrhages appeared as concentric ring structures with white punctate centers. The vitreal hemorrhage was characterized by the presence of choroidal blood in the vitreal chamber at the exposure site. The 30-nsec contained hemorrhage threshold (ED50) was 1.7 mJ on the macula and 2.1 mJ for an extramacular exposure. The 30-nsec vitreal hemorrhage macular threshold was 2.3 mJ, and the extramacular threshold was 6.6 mJ. The threshold for the 4-nsec pulsewidths to produce a hemorrhage (vitreal or contained) on the retina (macula or extramacular) was 340 microJ.

Bone density changes during two years treatment with tibolone or conjugated estrogens and norgestrel, compared with untreated controls in postmenopausal women.

OBJECTIVE: To quantify and compare the effect of two different hormone replacement therapy regimens on bone mineral density (BMD) and serum alkaline phosphate levels against a group of no treatment control volunteers. METHODS: An open 104-week comparative study of 113 early postmenopausal women. Fifty volunteered for the no treatment arm; the remainder were randomized to tibolone (T) (n = 32) or conjugated estrogens plus sequential norgestrel (CEEP) (n = 31). BMD was measured at baseline and after 48 and 96 weeks by dual photon absorptiometry. RESULTS: Baseline BMD was lower in nontherapy controls and with women assigned to CEEP compared with the T group. Statistical significance was not reached. By 96 weeks, increased BMD was observed with both therapies at all sites, whereas controls had lost bone compared with baseline. Changes from baseline with both preparations were significantly different (p < 0.05) from changes in controls for all sites, except femoral neck where the change was significant only with CEEP (p < 0.05). Alkaline phosphatase was significantly reduced in both treatment groups compared with controls (p < 0.05) at 48 and 96 weeks. CONCLUSIONS: The significant increases in BMD with T and CEEP confirm both regimens are effective in preventing osteoporosis. Without treatment, BMD declines postmenopausally.

Portal lymphadenopathy associated with lipofuscin in chronic cholestatic liver disease.

To determine whether portal lymphadenopathy in primary biliary cirrhosis is caused by deposition of lipofuscin pigment in sinus histiocytes and to compare primary biliary cirrhosis with other liver diseases a retrospective study on a consecutive series of 169 livers obtained at transplantation was carried out. There were grouped into eight diagnostic categories: primary biliary cirrhosis (n = 51), primary sclerosing cholangitis (n = 10), extrahepatic biliary atresia (n = 6), chronic rejection (n = 9), cirrhosis (other causes) (n = 38), primary liver neoplasia (n = 21), acute liver disease (n = 20), and retransplantation (other) (n = 14). Lymph nodes were present in 66 specimens. Fifty of these contained granules of lipofuscin pigment. The highest incidence of lymph node enlargement and the largest amounts of pigment were present in cases of primary biliary cirrhosis. A similar pattern of lymph node enlargement was also commonly observed in other chronic cholestatic conditions (primary sclerosing cholangitis, biliary atresia, chronic rejection). Much less pigment was seen in nodes draining livers with non-cholestatic cirrhosis or primary tumours. Nodes were not found in acute liver disease. It is concluded that portal lymphadenopathy associated with lipofuscin is a common finding in various chronic cholestatic liver diseases. The pathogenesis of this lesion is uncertain. Most cases are asymptomatic with enlarged nodes which may be detected only at laperotomy or necropsy and may be wrongly attributed to neoplastic disease. Diagnostically, the finding of large amounts of lipofuscin in enlarged portal lymph nodes is a good indicator of underlying chronic cholestatic liver disease.

Discriminant histological features in the diagnosis of chronic idiopathic inflammatory bowel disease: analysis of a large dataset by a novel data visualisation technique.

BACKGROUND/AIMS: The histopathological assessment of endoscopic colorectal biopsies is important in the distinction between normality and chronic idiopathic inflammatory bowel disease, and between ulcerative colitis and Crohn's disease, in subjects with symptoms of bowel dysfunction. This study aims to use carefully defined histopathological observations on a large study population to produce systems that improve classification into these diagnostic categories. METHODS: Eight hundred and nine endoscopic colorectal biopsies with verified outcomes (165 normal, 473 ulcerative colitis, 171 Crohn's disease) were examined by a single experienced histopathologist and 20 defined features were recorded for each case using a novel graphical interface with reference images of each feature. These features, together with age and sex, were used to produce and test statistical classifiers using logistic regression and a novel growing cell structure technique. RESULTS: The distinction between chronic idiopathic inflammatory bowel disease and normality was made with a good level of performance by both statistical classifiers (with areas under the receiver operating characteristic curves above 0.80). The growing cell structure system selected features as discriminant that agreed with the published literature. Logistic regression produced a more variable selection of discriminant features because of the high correlation between many features. The distinction between ulcerative colitis and Crohn's disease was performed less accurately, with areas under the receiver operating characteristic curves of about 0.70. Again the features selected as discriminant broadly agreed with those in the published literature. CONCLUSIONS: Histopathological examination of endoscopic colorectal biopsies is an effective method of distinguishing between subjects with chronic idiopathic inflammatory bowel disease and normality, but less good at distinguishing between ulcerative colitis and Crohn's disease. The features selected as discriminant in this large statistical analysis broadly agree with those published in the literature from more qualitative studies.

Cytokeratin immunostaining for detection of biliary epithelium: its use in counting bile ducts in cases of liver allograft rejection.

AIMS: To see how useful the application of a bile duct specific cytokeratin antibody (AE1) was in identifying and counting bile ducts in liver allograft biopsy specimens. METHODS: Eighteen liver biopsy specimens showing acute rejection and 17 biopsy specimens plus six hepatectomy specimens showing chronic rejection were studied. Serial sections were cut and stained with haematoxylin and eosin and AE1 antibody. Two pathologists (RFH and KP) examined the sections with respect to a range of histological features. RESULTS: Similar numbers of bile ducts were identified on haematoxylin and eosin sections as on corresponding sections stained by AE1 in cases of acute rejection and end stage chronic rejection. Greater numbers of bile ducts were identified by AE1 during the early stages of chronic rejection, especially when dense portal inflammatory infiltrates were present. These were often incomplete structures or individual cells within portal tracts, and bile ducts subsequently disappeared in all cases. Ductular proliferation was clearly shown by AE1 in acute rejection and the extent seemed to correlate with the severity of rejection present. By contrast, no ductular proliferation was observed in chronic rejection. CONCLUSIONS: Haematoxylin and eosin stained sections are adequate for counting bile ducts in most biopsy specimens from patients with suspected chronic rejection. Immunostaining for biliary cytokeratins using AE1 is of limited use in occasional cases where bile ducts are obscured by inflammatory cells.

The metabolic marker tumour pyruvate kinase type M2 (tumour M2-PK) shows increased expression along the metaplasia-dysplasia-adenocarcinoma sequence in Barrett's oesophagus.

BACKGROUND: Proliferating and tumour cells express the glycolytic isoenzyme, pyruvate kinase type M2 (M2-PK). In tumours cells, M2-PK usually exists in dimeric form (tumour M2-PK), causing the accumulation of glycolytic phosphometabolites, which allows cells to invade areas with low oxygen and glucose concentrations. AIMS: To investigate the expression of tumour M2-PK during the metaplasia-dysplasia-adenocarcinoma sequence of Barrett's oesophagus, and to assess the prognostic usefulness of tumour M2-PK in oesophageal cancer. MATERIALS/METHODS: One hundred and ninety cases selected from the histopathology archives as follows: 17 reflux oesophagitis, 37 Barrett's oesophagus, 21 high grade dysplasia, 112 adenocarcinomas, and three control tumours. Sections were stained immunohistochemically with antibody to tumour M2-PK. RESULTS: Tumour M2-PK was expressed in all cases, and increased cytoplasmic expression was seen with progression along the metaplasia-dysplasia-adenocarcinoma sequence. All cases of adenocarcinoma showed 100% staining so that tumour M2-PK was not a useful prognostic marker. CONCLUSIONS: Tumour M2-PK is not a specific marker of Barrett's adenocarcinoma, but may be important as a marker of transformed and highly proliferating clones during progression along the metaplasia-dysplasia-adenocarcinoma sequence.

Can women with gonadotropin levels diagnostic of polycystic ovarian syndrome benefit from therapy with dopamine agonists?

On the basis of LH and FSH data the incidence of PCOS in a series of 33 infertile women with hyperprolactinemia due to differing causes was found to be 39%. Where the elevations of prolactin were intermittent (spikers), the incidence was 44%, compared to 33% in a control population and 23% (9% histologically confirmed) in a 1991 Dublin general infertility clinic population. Androgen levels were uninformative, as were TRH stimulation test results. All 33 infertile women were treated with dopamine agonists. Four pregnancies occurred in the PCOS group. Two were hyperprolactinemic spikers on dopamine agonists plus antiestrogens. Four of the 10 pregnancies in the non-PCOS group were also on dopamine agonists plus antiestrogens. Two of these were spikers. The use of dopaminergic drugs to lower circulating prolactin is established. The addition of an antiestrogen in a concomitant PCOS situation is rational. There appears to be little justification, however, to use them in any anovulatory situation, including PCOS in the absence of hyperprolactinemia.

Assessment of factors constituting an "inducibility profile".

A survey of 440 patients examined vaginally during the last month of pregnancy showed a correlation between quantitative assessment of the pelvic score features described by Bishop and the duration of induced labor. Cervical dilatation was as useful as the total Bishop score in this respect. Cervical dilatation and station of the head together were especially predictive for long labor and these two factors together with length of cervix were especially predictive for shorter labors. It was also apparent from this study that previous cervical surgery or vaginal termination of pregnancy was associated with a lower incidence of low Bishop scores. The changes in individual components of the Bishop score during the last month of pregnancy are described.