Seminal plasma metabolomics profiles following long (4-7 days) and short (2 h) sexual abstinence periods.

OBJECTIVE: Metabolomic profiling of seminal plasma has been suggested as a possible approach for a fast and non-invasive male infertility evaluation diagnosis. However, metabolomics profiles in normozoospermic men have not been thoroughly investigated, and the influence of ejaculation-abstinence has not been described. To provide interim reference values and find associations between the metabolomics profiles of human seminal plasma and length of ejaculation-abstinence period in normozoospermic men. STUDY DESIGN: Semen samples collected after long (4-7 days) and short abstinence (2 h) from 31 normozoospermic males were assessed for routine quality parameters before the seminal plasma was separated by centrifugation. Metabolomics profiles of the seminal plasma were then determined using untargeted Nuclear Magnetic Resonance Spectroscopy. RESULTS: In total, 30 metabolites were identified. Pyruvate showed a higher concentration, while fructose, acetate, choline, methanol, N-acetylglucosamine, O-acetylcarnitine, uridine, and sn-glycero-3-phosphocoline showed lower concentrations in samples collected after short abstinence (vs. long). All metabolites showed lower absolute amounts (volume × concentration) following shorter abstinence. However, the lower sperm concentration in samples collected after short abstinence resulted in higher absolute amounts of pyruvate and taurine per spermatozoa: pyruvate 1.92 (1.12-3.87) vs. 1.29 (0.83-2.62) (P < 0.001) and taurine 0.58 (0.36-0.92) vs. 0.43 (0.28-0.95) (P < 0.05) ng/106 spermatozoa. Simultaneously, there was a higher percentage of progressively motile spermatozoa in samples collected after the short abstinence. CONCLUSION: The generally lower concentrations of seminal metabolites after short abstinence periods may be related to the shorter time available for secretion and collection of these metabolites by the accessory glands and the epididymides. The concomitant lower number of spermatozoa in the second ejaculate resulted in increased absolute amounts of pyruvate and taurine per spermatozoa, accompanied by increased spermatozoa motility in these samples. The simultaneous increase in percentages of motile spermatozoa and absolute amounts of pyruvate and taurine per spermatozoa after shorter abstinence might indicate that these two metabolites play a more critical role in sperm motility, which should be further investigated in future studies.

Improved sperm kinematics in semen samples collected after 2 h versus 4-7 days of ejaculation abstinence.

STUDY QUESTION: Does a short abstinence period of only 2 h yield spermatozoa with better motility characteristics than samples collected after 4-7 days? SUMMARY ANSWER: Despite lower semen volume, sperm concentration, total sperm counts and total motile counts, higher percentages of motile spermatozoa with higher velocity and progressiveness were detected in samples obtained after 2 h. WHAT IS KNOWN ALREADY: Most studies that have assessed the effect of abstinence periods on sperm motility parameters in men with a sperm concentration below 15 million/ml have detected a higher percentage of motile spermatozoa in samples obtained after short abstinence periods. Studies of men with sperm concentrations above 15 million/ml have reported significantly decreased motile sperm counts after 24 h of abstinence compared with longer abstinence periods. STUDY DESIGN, SIZE, DURATION: This study had a controlled repeated-measures design based on semen samples from 43 male partners, in couples attending for IVF treatment, who had a sperm concentration above 15 million/ml. Data were collected between June 2014 and December 2015 in the Fertility Unit of Aalborg University Hospital (Aalborg, Denmark). PARTICIPANTS/MATERIALS, SETTING, METHODS: Participants provided a semen sample after 4-7 days of abstinence followed by another sample after only 2 h. For both ejaculates, sperm concentration, total sperm counts, motility groups and detailed kinematic parameters were assessed and compared by using the Sperm Class Analyzer (SCA) computer-aided sperm analysis system before and after density gradient selection. The laboratory's local manual method (Makler chamber) was used for comparison. MAIN RESULTS AND THE ROLE OF CHANCE: The second raw ejaculate demonstrated lower semen volume (P < 0.0001), sperm concentration (P = 0.003) and sperm counts in all motility sub-groups (P < 0.001) but higher percentages of spermatozoa with higher velocity (P < 0.01), progressiveness (P < 0.001) and hyperactivation (P < 0.001), compared with the first raw ejaculate. LIMITATIONS, REASONS FOR CAUTION: The first ejaculate in this study was also used for the IVF/ICSI treatments and therefore only patients with a semen volume ≥2 ml and concentration ≥15 million/ml were included. Further validation in large prospective randomized controlled trials, more purposely directed at normozoospermic males with partners having problems conceiving when there appears to be no female factor, is needed to confirm the potential advantage of using a second semen sample in improving fertilization and pregnancy rates in assisted reproduction. WIDER IMPLICATIONS OF THE FINDINGS: Despite the significantly lower semen volume, sperm concentration and total sperm counts in all motility sub-groups, the significantly higher percentage of spermatozoa with better motility characteristics (velocity, progressiveness and hyperactivation) in the second ejaculate, may provide and allow for a simpler and more effective selection of higher quality spermatozoa. This could prove to be an advantage for ART procedures such as intracytoplasmic sperm injection where a large number of spermatozoa is not needed. It can also be speculated that pooling two consecutive ejaculates obtained after 4-7 days and after 2 h, could be an advantage for intrauterine insemination where a large number of motile spermatozoa are needed. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by internal grants from the Department of Health Science and Technology, Faculty of Medicine, Aalborg University (Aalborg, Denmark). The SCA® was provided by a grant from 'Ferring Pharmaceuticals' to Aalborg University Hospital (H.I.N). G.V.D.H. is an external senior scientific consultant to Microptic S/L (Barcelona, Spain). H.A. has provided scientific input and presentations for Microptic S/L (Barcelona, Spain) on several occasions. All other authors declare no conflict of interest. TRIAL REGISTRATION NUMBER: N/A.

Impact of hepatitis B virus co-infection on response to highly active antiretroviral treatment and outcome in HIV-infected individuals: a nationwide cohort study.

BACKGROUND: The impact of chronic hepatitis B virus (HBV) infection on viral suppression, immune recovery and mortality in HIV-1 infected patients on highly active antiretroviral treatment (HAART) is a matter of debate. The impact of HBeAg status is unknown. METHODS: This prospective cohort study included all adult Danish HIV-1 infected patients who started HAART between 1 January 1995 and 1 December 2006 (3180 patients). Patients were classified as chronic HBV-infected (6%), HBV-negative (87%) or HBV-unknown (7%). HBV-positive patients were divided into HBeAg-positive or -negative (3.0 vs. 2.6%). Study endpoints were viral load, CD4 cell count and mortality. RESULTS: HBV co-infection had no impact on response to HAART regarding viral suppression or immune recovery. HBV co-infection was associated with several outcomes: overall mortality [mortality rate ratio (MRR) 1.5; 95% confidence interval (CI) 1.1-2.1], liver-related mortality (MRR 4.0; 95% CI 1.6-9.9) and AIDS-related deaths (MRR 1.7; 95% CI 1.0-3.0). The presence of HBeAg did not influence patients' response to HAART. CONCLUSIONS: In HIV patients, chronic HBV infection has no impact on response to HAART concerning viral load and increase in CD4 cell count. However, co-infected patients have an increased mortality compared to HIV-monoinfected patients.

IVM--the first choice for IVF in Italy.

In March 2004, a new law was introduced in Italy to regulate assisted reproduction; at present it is impossible to use more than a maximum of three oocytes per IVF cycle, nor can embryos or prezygotes (2PN cells) be selected or cryopreserved. The prohibitions introduced by the new law have, on the one hand, reduced the expectations of success of current techniques and, on the other hand, stimulated clinicians and embryologists to work on new therapeutic strategies so as to offer the highest chances of success with the lowest risks. In-vitro maturation (IVM) of oocytes fits very well with these new requirements: ovarian stimulation is avoided and the handling of spare oocytes is facilitated. The IVM protocol is an intriguing alternative to conventional IVF techniques, since it removes the side-effects of drug stimulation, especially ovarian hyperstimulation syndrome, and it also reduces the costs of the entire procedure, both in terms of 'time consumption' and 'patient/society costs for drugs'. In the authors' IVF centre the IVM technique has been used for more than a year, with significant success in terms of maturation and fertilization rates, percentage of embryo transfers, number of pregnancies and, finally, healthy babies born.

[Use of antiretroviral chemoprophylaxis after occupational HIV exposure]. / Anvendelse af antiretroviral kemoprofylakse efter erhvervsbetinget eksposition for HIV.

The study describes the use of postexposure antiretroviral prophylaxis (PEP) after occupational exposure to HIV in Denmark in the period 1/1 1997-31/12 1998. Forty-seven accidentally exposed persons received PEP in this period, of whom 14 were nurses, 13 physicians and 11 were not employed in the health care system. In 23 cases side effects to PEP were described. In 18 cases the index person was i.v. drug abuser and in 11 cases homosexual/bisexual men. In 18 cases the HIV status of the index person was unknown at the time of exposure. Ten of these index persons were subsequently tested and all found HIV-negative. It is concluded, that the frequency of occupational exposure to HIV is unacceptably high. Further it is emphasised that in cases where the HIV status of the index person is unknown, PEP should only be instituted if the index person is at risk of being HIV infected and has signs of HIV infection.

[Clinical effect of ranitidine in psoriasis. An open prospective study]. / Klinisk effekt af ranitidin ved psoriasis. Et abent prospektivt studie.

We report the results of an open, prospective study of 20 patients with moderate to severe psoriasis (median PASI score: 15.7) treated with oral ranitidine 300 mg twice daily for six months. No other medication was allowed during the study period. The median PASI score was reduced to 14.5, 9.1 and 5.7 after one, three and six months of treatment, respectively (p < 0.00001). A mild to moderate worsening was observed in 15 patients within the first month of treatment, but this was not followed by exclusion from the study, and long-term treatment improved disease status in most patients. Eight patients continued therapy with ranitidine 300 mg twice daily after the study was completed. None of these patients relapsed during a follow-up period of 12-18 months. The results of the present study suggest that ranitidine may be beneficial in the treatment of some patients with psoriasis.

[The effect of ranitidine on postoperative monocyte and neutrophil granulocyte function]. / Effekten af ranitidin på den postoperative monocyt- og neutrofile granulocytfunktion.

The histamine H-2-receptor antagonist ranitidine hydrochloride has been shown to alleviate trauma-, blood transfusion- and sepsis-induced immunosuppression. We evaluated the effect of ranitidine on the postoperative impairment of monocyte and neutrophil function in 24 patients undergoing major elective abdominal surgery. The patients were randomized to receive postoperative adjuvant treatment with ranitidine hydrochloride (100 mg) administered intravenously twice daily for four days, followed by oral ranitidine hydrochloride (150 mg) administered twice daily for five days (n = 11), or no adjuvant treatment (n = 13). Blood monocyte and neutrophil chemotaxis and chemiluminescence were analyzed before the operation and on post-operative days one, three and nine. Monocyte chemotaxis to C5a in the 13 control patients was significantly decreased on day one compared to day 0. Chemotaxis in the 11 ranitidine-treated patients increased significantly from day 0 to day one (p < .01 between groups). Neutrophil chemiluminescence to zymosan and N-f-methionyl-leucylphenylalanine was significantly increased in control patients on day one compared to day 0 (p < .05), while ranitidine reduced chemiluminescence to zymosan insignificantly on day one (p < .07 between groups). Five of the 13 control patients developed postoperative infectious complications, which were related to decreased monocyte chemotaxis to C5a and increased neutrophil chemiluminescence to zymosan when compared to noninfected patients. A significant difference (P < .05) in chemiluminescence to zymosan between infected and noninfected control patients was observed on day three, before clinical signs of infectious disease could be detected. There were no infectious complications in ranitidine-treated patients. These results support previous studies on the effect of ranitidine in reducing postoperative immunosuppression.

Optimization and simplification of culture conditions in human in vitro fertilization (IVF) and preembryo replacement by serum-free media.

The results of 220 consecutive IVF treatments are presented, comparing the use of culture media supplemented with either patient serum (Group 1; n = 110), or Medi-Cult SSR 2 synthetic serum replacement with pyruvate, and human serum albumin (HSA) (GEA BioTech, Hvidovre, Denmark) (Group 2; n = 110). In both groups the Medi-Cult Hybritest was used for routine quality testing. A significantly (P less than 0.05) increased rate of deliveries/ongoing pregnancies was observed with the Group 2 medium. However, no significant differences in fertilization rate, cleavage rate, or implantation rate were observed. It is concluded that the serum-free culture medium described and the testing for absence of cytotoxicity in a sensitive bioassay (Hybritest) have yielded culture conditions capable of sustaining the development in vitro of human preembryos without impairing the fertilization process or the implantation rate, ultimately resulting in a significantly increased rate of deliveries/ongoing pregnancies and an apparently decreased abortion rate. The potential harmful effects of serum and the need for blood sampling and preparation further increase the advantages of replacing serum with the synthetic serum replacement SSR 2 in an IVF program.

Culture of normal adult human melanocytes.

Melanocytes isolated from normal adult human skin were cultured in vitro. Separation of the epidermis from the dermis by trypsin flotation proved better than collagenase treatment for providing viable cultures of melanocytes with a minimum of fibroblast contamination. Centrifugation on a discontinuous, rather than a continuous Percoll gradient, was more efficient in separating the epidermal cell types. Most of the melanocytes were usually found in one particular layer, and most of the viable keratinocytes were in the sediment. None of the layers produced a uniformly high percentage of melanocytes on routine culture, but enriched melanocyte cultures could be obtained by seeding the epidermal cells in magnesium- and calcium-free medium for 24 to 48 hours, and then transferring them to fibroblast-conditioned medium containing horse serum and polyamines. Melanocytes were identified by their dendritic morphology, ultrastructure, reaction to cholera toxin and pigment production after treatment with melanocyte stimulating hormone. Pure cultures of melanocytes have been cultivated by this method for more than 43 weeks (ten passages).

Ultrastructural changes in the dermal chromatophore unit of Hyla arborea during color change.

The structural changes in the chromatophores of Hyla arborea related to changes in skin color were studied by electron microscopy and reflectance microspectrophotometry. During a change form a light to a darker green color, the melanosomes of the melanophores disperse and finally surround the iridophores and partly the xanthophores. The iridophores change from cup-shape to a cylindrical or conical shape with a simultaneous change in the orientation of the platelets from being parallel to the upper surface of the iridophores to being more irregular. The xanthophores change from lens-shape to plate-shape. The color change from green to grey seems always to go through a transitional black-green or dark olive green to dark grey. During this change the xanthophores migrate down between the iridophores, and in grey skins they are sometimes found beneath them. The pterinosomes gather in the periphery of the cell, while the carotenoid vesicles aggregate around the nucleus. The iridophores in grey skin are almost ball-shaped with concentric layers of platelets. A lighter grey color arises from a darker grey by an aggregation of melanosomes. The chromatophore values previously defined for Hyla cinerea are applicable in Hyla arborea, and the ultrastructural studies support the assumptions previously made to explain these values.