An algorithm to predict pregnancy in assisted reproduction

BACKGROUND: Male fertility potential cannot be measured by conventional parameters for the assisted reproduction technique; ICSI. This study determines the relationship between testicular and ejaculated sperm mitochondrial (mt) DNA deletions, nuclear (n) DNA fragmentation, and fertilization and pregnancy rates in ICSI. METHODS: Ejaculated sperm were obtained from 77 men and testicular sperm from 28 men with obstructive azoospermia undergoing ICSI. Testicular sperm were retrieved using a Trucut needle. mtDNA was analysed using a long PCR. The alkaline Comet assay determined nDNA fragmentation. RESULTS: Of subjects who achieved a pregnancy (50%) using testicular sperm, only 26% had partners' sperm with wild-type (WT) mtDNA. Of pregnant subjects (38%) using ejaculated sperm, only 8% had partner sperm with WT mtDNA. In each, the successful group had less mtDNA deletions and less nDNA fragmentation. There were inverse relationships between pregnancy and mtDNA deletion numbers, size and nDNA fragmentation for both testicular and ejaculated sperm. No relationships were observed with fertilization rates. An algorithm for the prediction of pregnancy is presented based on the quality of sperm nDNA and mtDNA. CONCLUSION: In both testicular and ejaculated sperm, mtDNA deletions and nDNA fragmentation are closely associated with pregnancy in ICSI.     

Screening assays for immunologic infertility: a comparison study.

Because the immunobead antisperm antibody test (IBT) is difficult to learn and results are heavily operator-dependent, we compared findings on IBT with those of SpermMAR and SpermCheck in 58 patients to determine whether the latter two tests could demonstrate improved screening ability. Assays performed on both sperm and serum (i.e., direct and indirect) yielded 100% positive predictive values when IBT was used as the standard. Both SpermCheck and SpermMAR are easier to perform, and, as the former has a slightly greater sensitivity, we recommend SpermCheck for the detection of immunologic infertility.     

Detection of Pseudomonas aeruginosa producing metallo-beta-lactamases in a large centralized laboratory

Metallo-beta-lactamases (MBLs) have been increasingly recognized from clinical isolates worldwide, but the laboratory detection of these strains is not well defined. We report a study that developed an EDTA disk screen test and a molecular diagnostic assay for the detection of MBL-producing Pseudomonas aeruginosa. Using NCCLS disk methodology, inhibition zone diameters were determined in tests with imipenem (IPM) and meropenem (MEM) disks alone and in combination with 930 microg of EDTA. This test was compared with the MBL Etest. The duplex PCR assay showed 100% sensitivity and specificity for detecting MBL-producing control strains. Of the 241 clinical strains of IPM-nonsusceptible P. aeruginosa from the Calgary Health Region isolated from 2002 to 2004, 110/241 (46%) were MBL positive using phenotypic methods while 107/241 (45%) were PCR positive for MBL genes: 103/241 (43%) for bla(VIM) and 4/241 (2%) for bla(IMP). The EDTA disk screen test using MEM showed 100% sensitivity and 97% specificity for detecting MBLs in control and clinical strains. The EDTA disk screen test is simple to perform and to interpret and can easily be introduced into the workflow of a clinical laboratory. We recommend that all IPM-nonsusceptible P. aeruginosa isolates be routinely screened for MBL production using the EDTA disk screen test and that PCR confirmation be performed at a regional laboratory.     

Analysis of cell division among subpopulations of lymphoid cells in sheep. I. Thymocytes

The number, distribution and surface phenotype of dividing cells in the thymus, and differences between the cell cycle status of thymocyte subpopulations, were studied in fetal and post-natal lambs using double-labelling techniques. Dividing cells were labelled in vivo for various periods with 5-bromo-2-deoxyuridine (BrdU). The proportions of constituent thymocyte subpopulations that had synthesized DNA during the labelling period were measured by flow cytometry or immunohistochemistry using a panel of monoclonal antibodies (mAbs) specific for sheep lymphocyte differentiation antigens and MHC class I and class II antigens in conjunction with an anti-BrdU mAb. The proportion of thymocytes that incorporated BrdU during a 1-hr labelling period varied with age, and levels of 30%, 13% and 9% were measured, respectively, in 40- and 125-day-old fetuses and 8-week-old lambs. Eight percent of the thymocytes in lambs were synthesizing DNA, with 4% entering the G2 phase per hour, and a substantial number of thymocytes (21%) had a G2 + M phase DNA content. A small subset of thymocytes (1-3%) recognized by mAb E-79 localized to the subcapsular region of the cortex and displayed the highest level of BrdU incorporation. Cortical-type thymocytes (CD1+) comprised 50-70% of thymocytes; however, few of these incorporated BrdU and the proportion in the G2 + M phase of the cell cycle was higher than for other thymocyte subpopulations. The 197+CD4-CD8- T cells also showed no evidence of in vivo division.     

The effects of communication skills training on patients' participation during medical interviews

Recent models of physician-patient communication emphasize information exchange in promoting partnership. Although considerable attention has been given to physicians' information exchange, little research has examined patients' communication contributions. The purpose of this research was to test the effectiveness of a training booklet designed to enhance patients' communication skills in information exchange. A nested design was used, such that 25 physicians each saw six patients, two patients in each of three communication skills interventions (i.e. trained, informed, control). The dependent variables included several discourse categories designed to assess patients' information seeking, provision, and verifying. Results indicate that trained patients engaged in more effective and efficient information seeking, provided physicians with more detailed information about their medical condition, and used more summarizing utterances to verify information they received from physicians. Additionally, dyads consisting of trained patients demonstrated a more patient-controlled style of communication than did dyads consisting of informed or untrained patients.     

Localisation of aluminium by histochemical and electron probe x-ray microanalytical techniques in bone tissue of cases of renal osteodystrophy.

Histochemical studies and electron probe x-ray microanalysis for aluminium have been performed on 16 samples of undecalcified bone from cases of renal osteodystrophy associated with a syndrome suggestive of dialysis encephalopathy (five cases), age and sex matched controls for these and a group of patients with chronic renal failure (six cases) who have never been on haemodialysis. Aluminium was detected only in the patients with a dialysis encephalopathy-like syndrome. This group had significant histological bone disease the features of which were broadly consistent with the so-called atypical renal osteomalacia which is thought to be due to a metal toxin. Aluminium was demonstrated at the interface between osteoid and mineralised tissue--that is, at the site of the calcification front, where it could interfere with the mineralisation process. In the group of patients who had never been subjected to haemodialysis there was also significant histological bone disease but no evidence of aluminium accumulation. In this group the bone disease was of a more typical pattern of osteomalacic changes coupled with those of hyperparathyroidism.     

Semen parameters and testicular pathology in men with testicular cancer and contralateral carcinoma in situ or bilateral testicular malignancies

We evaluated 14 patients with bilateral testicular tumour, one-sided tumour and contralateral carcinoma in situ (CIS) of the testis or testis tumour in single testis with respect to their fertility. We analysed semen parameters, serum hormones [follicle-stimulating hormone (FSH), luteinizing hormone (LH) and testosterone], testicular sonography, testicular volumes and testicular histology prior to further anti-cancer treatment. Ten out of 14 patients showed normal or reduced sperm concentrations, while 4/14 patients were azoospermic. Serum FSH levels showed a significant negative correlation with sperm concentrations in patients with testicular malignancies (r = -0.64, P = 0.025). Testicular volumes revealed a significant positive correlation with semen parameters in patients with testes that were affected by CIS (r = 0.733, P = 0.038). We conclude that even bilateral testicular cancer and/or CIS do not preclude fertility and, therefore, patients should be offered andrological investigation and therapy, including possibly surveillance strategy or the chance for cryopreservation of the semen prior to further treatment in order to preserve their chances for paternity.      

Studies using a viral challenge and CD8 T cell depletions on the roles of cellular and humoral immunity in the control of an SHIV-89.6P challenge in DNA/MVA-vaccinated macaques

Here, we study immune responses in four DNA/MVA-vaccinated macaques following an SHIV-89.6P challenge and a subsequent CD8 cell depletion. Both post-challenge and post-depletion peaks of viremia contracted with the expansion, or re-emergence, of CD8 T cells. Post-depletion, CD8 cells expanded in the presence of higher levels of neutralizing Ab and CD4 help than post-challenge and had superior maturational characteristics as measured by expression of the anti-apoptotic protein Bcl-2, the IL-7 receptor CD127 and co-production of IFN-gamma and IL-2. Pre-challenge and pre-depletion titers of neutralizing Ab correlated inversely with peaks of viremia and directly with peaks for anti-viral CD4 cells. Thus, our results reveal CD8 cells playing a central role, and neutralizing Ab, a supporting role in SHIV-89.6P control. They also suggest a dynamic relationship between neutralizing Ab, antigen load and anti-viral CD4 cells in the maturation of high-quality anti-viral CD8 T cells.