Moderate drinking : No impact on female fecundity

Objective: To determine the effect of moderate alcohol intake on probability of conception. Design: Prospective cohort study. Setting: Normal healthy women enrolled in an artificial donor insemination program. Patients: Women starting artificial donor insemination for the very first time were selected. Interventions: Alcohol exposure was measured through a self- administered questionnaire at intake of study. Main Outcome Measure: A first positive pregnancy test as indicative for conception. Results: Women with moderate alcohol intake had a slightly higher, though not significant, probability of conception compared with nondrinkers (Hazard Ratio 1.20; 95% confidence interval 0.90 to 1.60). Conclusion: Moderate alcohol intake has no negative impact on female fecundity

Injectable hydrogels functionalized with antibody fragments as intra-articular cytokine sinks neutralizing pro-catabolic cytokines

Purpose: Intra-articular (IA) injection with biologics like cytokine neutralizing antibodies to treat osteoarthritis (OA) have been explored, but so far success is limited. One of the key obstacle for the efficacy of intra-articular therapies is the short retention time of these biologics in the joint space due to rapid clearing by the synovial membrane. To overcome these limitations, we developed a new concept that potentially can increase the therapeutic value of pro-inflammatory cytokine neutralizing antibody fragments in the management of OA. The concept is based on the use of injectable hydrogels functionalized with the variable domain of single chain heavy chain only antibodies (VHHs) for cytokine capture. VHHs can be easily isolated using phage-display technology, recombinantly engineered and produced in yeast. These hydrogels can either be pre-crosslinked in microgels before injection or injected as polymer-VHH conjugates that cross link in a hydrogel in situ. Here, hyaluronic acid was used as model polymer after substitution with tyramine and maleimide groups. The tyramine moiety is used in an enzymatic cross-linking reaction to yield stable macromolecular hydrogel networks. The maleimide group is used for thiol-maleimide chemistry coupling a recombinantly engineered VHH with a free Cysteine in its C-terminal tail to the polymer backbone. We hypothesized that this strategy to functionalize hydrogels with the VHH can retain biological activity of neutralizing VHH. To proof the concept of these so-called cytokine sinks we have selected an anti-TNFα VHH that effectively neutralizes TNFα, which plays an essential role in cartilage degradation in inflammatory arthritis. Methods: To achieve directed conjugation of the VHH to the polymer, we have introduced an unpaired cysteine by using recombinant DNA technology in the C-terminus of the anti-TNFα VHH. Afterwards, the modified VHH was produced in yeast and purified. The conjugation of the VHH to the hyaluronic acid was performed by incubating the VHH with tyramine and maleimide functionalized hyaluronic acid. Characterization of binding affinity for both the non-conjugated VHH and conjugated VHH were measured by ELISA. hydrogels functionalized with the VHH was prepared by mixing the hyaluronic acid-VHH conjugates with enzyme peroxidase and H2O2. Biological activity of the conjugated VHH and hydrogels functionalized with the VHH was measured using an NFκb responsive luciferase reporter cell line after stimulation with TNFα. An equimolar concentration of non-conjugated VHH was used as a control. Results: We show successful conjugation of the VHH to the polymer backbone. The VHH functionalized polymer could be used for making stable hydrogels after tyramine mediated cross linking. This modification did not affect the biological activity of the VHH. Using an NFκb responsive luciferase reporter cell line we demonstrated that hydrogels functionalized with the VHH efficiently inactivated TNFα and that this inhibition was comparable to inhibition by an equimolar concentration of non-conjugated VHH. Conclusions: We successfully developed a biocompatible and efficient way to couple VHH to hyaluronic acid. These conjugates could be used for in situ generation of cytokine sinks capable of capturing different pro-catabolic cytokines in treatment of OA. Our Results demonstrate that cytokine sinks have great potential for neutralizing inflammatory pro-catabolic cytokines after intra-articular injection by increasing the retention time of neutralizing antibody fragments in the joint cavity

Fat and female fecundity: prospective study of effect of body fat distribution on conception rates

OBJECTIVES: To study the effect of body fat distribution in women of reproductive age on fecundity. DESIGN: Prospective cohort study of all women who had entered a donor insemination programme. SETTING: One fertility clinic serving a large part of the midwest of the Netherlands. SUBJECTS: Of 542 women attending the clinic for artificial insemination for the first time, 500 women were eligible for study. MAIN OUTCOME MEASURES: Probability of conception per cycle and number of insemination cycles before pregnancy or stopping treatment. RESULTS: A 0.1 unit increase in waist-hip ratio led to a 30% decrease in probability of conception per cycle (hazard ratio 0.706; 95% confidence interval 0.562 to 0.887) after adjustment for age, fatness, reasons for artificial insemination, cycle length and regularity, smoking, and parity. Increasing age was significantly related to lower fecundity (p < 0.05); very lean and obese women were less likely to conceive (p < 0.10) as were women with subfertile partners (p < 0.10). All other exposure variables were not significantly related to fecundity. CONCLUSIONS: Increasing waist-hip ratio is negatively associated with the probability of conception per cycle, before and after adjustment for confounding factors. Body fat distribution in women of reproductive age seems to have more impact on fertility than age or obesity

Delaying childbearing: effect of age on fecundity and outcome of pregnancy.

OBJECTIVES--To study the age of the start of the fall (critical age) in fecundity; the probability of a pregnancy leading to a healthy baby taking into account the age of the woman; and, combining these results, to determine the age dependent probability of getting a healthy baby. DESIGN--Cohort study of all women who had entered a donor insemination programme. SETTING--Two fertility clinics serving a large part of The Netherlands. SUBJECTS--Of 1637 women attending for artificial insemination 751 fulfilled the selection criteria, being married to an azoospermic husband and nulliparous and never having received donor insemination before. MAIN OUTCOME MEASURES--The number of cycles before pregnancy (a positive pregnancy test result) or stopping treatment; and result of the pregnancy (successful outcome). RESULTS--Of the 751 women, 555 became pregnant and 461 had healthy babies. The fall in fecundity was estimated to start at around 31 years (critical age); after 12 cycles the probability of pregnancy in a woman aged greater than 31 was 0.54 compared with 0.74 in a woman aged 20.31. After 24 cycles this difference had decreased (probability of conception 0.75 in women greater than 31 and 0.85 in women 20.31). The probability of having a healthy baby also decreased--by 3.5% a year after the age of 30. Combining both these age effects, the chance of a woman aged 35 having a healthy baby was about half that of a woman aged 25. CONCLUSION--After the age of 31 the probability of conception falls rapidly, but this can be partly compensated for by continuing insemination for more cycles. In addition, the probability of an adverse pregnancy outcome starts to increase at about the same age