Factors associated with embryo mosaicism: a systematic review and meta-analysis.

PURPOSE: Evaluate which factors are involved in the increased rate of mosaicism in embryos. METHODS: A systematic review and meta-analysis was performed. After an exhaustive search of the literature, a total of seven papers were included in the analysis. In addition, data collected from IVF cycles performed in our fertility clinic were also analysed. Day of biopsy, embryo quality, maternal and paternal age and seminal quality were the chosen factors to be studied. RESULTS: The results of the meta-analysis show that neither embryo quality nor seminal quality were related to mosaic embryo rate (OR: 1.09; 95% CI: 0.94-1.28 and OR: 1.10; 95% CI: 0.87-1.37, respectively). A positive association was observed for the variable "biopsy day" with embryos biopsied at day 6 or 7 having the highest rate of mosaicism (OR: 1.06; 95% CI: 1.01-1.11). In opposite to what happens with aneuploidy rate, which increases with maternal age, embryo mosaicism is higher in younger women (<34 years) rather than in older ones (≥34 years) (OR: 0.95; 95% CI: 0.92-0.98). However, for the "paternal age" factor, no association with mosaicism was found (OR: 1.04; 95% CI: 0.90-1.21). CONCLUSIONS: With the present study, we can conclude that the factors related to the presence of mosaicism in embryos are the embryo biopsy day and maternal age. The rest of the studied factors showed no significant relationship with mosaicism. These results are of great importance as knowing the possible causes leading to mosaicism helps to improve the clinical results of reproductive treatments.

Identification of novel candidate genes associated with meiotic aneuploidy in human embryos by whole-exome sequencing.

PURPOSE: To identify novel genetic variants responsible for meiotic embryonic aneuploidy. METHODS: A prospective observational cohort study that included 29 couples who underwent trophectoderm biopsies from 127 embryos and performed whole-exome sequencing (WES) between November 2019 and March 2022. Patients were divided into two groups according to the expected embryo aneuploidy rate based on maternal age. RESULTS: After variant filtering in the WES analysis of 58 patients/donors, five heterozygous variants were identified in female partners from the study group that had an impact on embryo aneuploidy. Additionally, a slowdown in embryo development and a decrease in the number of blastocysts available for biopsy were observed in the study group embryos. CONCLUSION: This study has identified new candidate genes and variants not previously associated with meiotic embryo aneuploidy, but which are involved in important biological processes related to cell division and chromosome segregation. WES may be an efficient tool to identify patients with a higher-than-expected risk of embryo aneuploidy based on maternal age and allow for individualized genetic counselling prior to treatment.

Identification of new variants and candidate genes in women with familial premature ovarian insufficiency using whole-exome sequencing.

PURPOSE: To identify candidate variants in genes possibly associated with premature ovarian insufficiency (POI). METHODS: Fourteen women, from 7 families, affected by idiopathic POI were included. Additionally, 98 oocyte donors of the same ethnicity were enrolled as a control group. Whole-exome sequencing (WES) was performed in 14 women with POI to identify possibly pathogenic variants in genes potentially associated with the ovarian function. The candidate genes selected in POI patients were analysed within the exome results of oocyte donors. RESULTS: After the variant filtering in the WES analysis of 7 POI families, 23 possibly damaging genetic variants were identified in 22 genes related to POI or linked to ovarian physiology. All variants were heterozygous and five of the seven families carried two or more variants in different genes. We have described genes that have never been associated to POI pathology; however, they are involved in important biological processes for ovarian function. In the 98 oocyte donors of the control group, we found no potentially pathogenic variants among the 22 candidate genes. CONCLUSION: WES has previously shown as an efficient tool to identify causative genes for ovarian failure. Although some studies have focused on it, and many genes are identified, this study proposes new candidate genes and variants, having potentially moderate/strong functional effects, associated with POI, and argues for a polygenic etiology of POI in some cases.

Platelet-Rich Plasma for the Treatment of Photodamage of the Skin of the Hands. / Plasma rico en plaquetas en el tratamiento del fotodaño cutáneo en las manos.

BACKGROUND: Platelet-rich plasma (PRP) provides growth factors that stimulate fibroblast activation and induce the synthesis of collagen and other components of the extracellular matrix. The objective of this study was to evaluate the effect of PRP in the treatment of photodamage of the skin of the hands. MATERIAL AND METHODS: Experimental study enrolling persons with photoaged skin on the dorsum of the hands (Glogau photoaging scale, type III, or Fitzpatrick wrinkle classification, type II) were included between August 2012 and January 2013. A histological comparison was made of skin biopsies taken before and after the application of PRP to the skin of the dorsum of the hands. RESULTS: The mean (SD) age of the 18 women enrolled was 47.9 (4.3) years. Histological analysis showed an increase in the number of fibroblasts (P<.001), number of vessels (P<.001), and collagen density (P=.27). These changes produced significant improvements in the Fitzpatrick wrinkle and elastosis scale (P<.001) and in the Glogau photoaging scale (P=.01). CONCLUSIONS: PRP induced a reduction in the manifestations of skin aging, including an improvement in wrinkles and elastosis.

The Effect of Oxidative Stress on Thawed Bulk-Sorted Red Deer Sperm.

The aims of this study were to assess the effects of the sex-sorting process on post-thaw sperm quality as well as on induced oxidative stress damage (H2 O2 0 mm = H000; H2 O2 50 mm = H050; H2 O2 100 mm = H100) and the protective action of reduced glutathione (GSH) and Trolox, when comparing sorted (BSS) and non-sorted (NS) red deer spermatozoa incubated at 37°C. Sperm samples from three stags were collected by electroejaculation and frozen. Immediately after thawing, sperm motility was higher (p < 0.05) for NS (59% ± 3.3) than BSS (36.9% ± 5.8) sperm. Furthermore, the percentage of apoptotic sperm was higher (p < 0.05) for BSS (21.6% ± 5.0) than NS sperm (14.6% ± 1.2). The presence of H2 O2 increased DNA damage in NS (H000 = 4.1% ± 0.9; H050 = 9.3% ± 0.7; and H100 = 10.9% ± 2.3), but not in BSS sperm. However, in the presence of oxidant, GSH addition improved (p < 0.05) sperm motility in both groups of sperm samples as compared to their controls (NS: 44.5 ± 4.8 vs 21.1 ± 3.9 and BSS: 33.3 ± 8.1 vs 8.9 ± 1.8). These results demonstrate that the sperm-sorting process induces sublethal effects, albeit selecting a sperm population with a chromatin more resistant to oxidative stress than that in non-sorted sperm. Moreover, addition of GSH at 1 mm may be a good choice for maintaining the quality of stressed sperm samples, unlike Trolox, which inhibited sperm motility.

Incidence of hip fracture in Barranquilla, Colombia, and the development of a Colombian FRAX model.

A FRAX model for Colombia was released June 30, 2010. This article describes the data used to develop the Colombian FRAX model and illustrates its features compared to other countries. Hip fracture cases aged 50 years or more who were referred to all hospitals serving the city of Barranquilla were identified prospectively over a 3-year period (2004-2006). Age- and sex-stratified hip fracture incidence rates were computed using the 2005 census. Present and future numbers of hip fracture cases in Colombia were calculated from the age- and sex-specific incidence and the national population demography. Mortality rates for 1999 were extracted from nationwide databases and used to estimate hip fracture probabilities. For other major fractures (clinical vertebral, forearm, and humerus), incidence rates were imputed, using Swedish ratios for hip to other major osteoporotic fracture, and used to construct the FRAX model. Incidence of hip fracture increased with age, more markedly in women than in men. Over all ages, the female to male ratio was 1.7. By extrapolation, there were estimated to be 7,902 new hip fracture cases (2,673 men, 5,229 women) in Colombia in 2010, which was predicted to increase to 22,720 cases (7,568 men, 15,152 women) in 2035. The 10-year probability of hip or major fracture was increased in patients with a clinical risk factor, lower BMI, female gender, a higher age, and a decreased BMD T score. The remaining lifetime probability of hip fracture at the age of 50 years was 2.5 and 4.7 % in men and women, respectively, which were lower than rates in a Mexican population (3.8 and 8.5 %, respectively) and comparable with estimates for Venezuela (2.4 and 7.5 %, respectively). The FRAX tool is the first country-specific fracture-prediction model available in Colombia. It is based on the original FRAX methodology, which has been externally validated in several independent cohorts. Despite some limitations, the strengths make the Colombian FRAX tool a good candidate for implementation into clinical practice.

IL-6/IL-10 and IL-1ß/IL-4 ratios associated with poor ovarian response in women undergoing in-vitro fertilization.

The aim of this work was to evaluate whether serum cytokines levels are associated with ovarian response in IVF. 149 patients were included in a retrospective study. Cytokines IL-2, IL-4, IL- 6, IL-8, IL-10, VEGF, IFNγ, TNFα, IL-1α, IL-1ß, MCP-1 and EGF were measured by sandwich immunoassay previously to ovarian stimulation. Performing hierarchical cluster analysis, we observed that the antral follicle count, the total oocytes recovered and the MII recovered are grouped in the same cluster as the cytokines IL-2-4-6-10-1α-1ß, IFNγ y TNFα. Then, we found that the ratio between IL and 6 and IL-10 was higher in low responder women (2.15 versus 1.55; p = 0.035). If we establish 0.9 as a cut-off for the IL-6/IL-10, we observed that above this value the risk of having a low response to ovarian stimulation was more than 3 times greater than below this value (22.9 % versus 6.0 %; p = 0.007). Also, the ratio IL-1ß/IL-4 was higher in patients with normal or suboptimal response (0.62 versus 0.34; p = 0.034) and any patient with low response had a value greater than 0.7 (p = 0.003). As a conclusion, the IL-6/IL-10 and IL-1ß/IL-4 ratios showed differences between normoresponder women and patients with low ovarian response.

Intermediate and normal sized CGG repeat on the FMR1 gene does not negatively affect donor ovarian response.

BACKGROUND: Fragile X syndrome is associated with low ovarian reserve and poor ovarian response. The aim of this study was to investigate whether CGG repeats on the fragile X mental retardation 1 (FMR1) gene have predictive value for ovarian response to stimulation with gonadotrophins and for clinical outcome in our oocyte donation program. METHODS: Oocyte donor candidates were selected according to Instituto Bernabeu oocyte donation program requirements. Fragile X genetic screening was performed in 204 oocyte donors, defining 141 controls and 63 cases: 35-39 repeats (n = 34), 40-45 (n = 12) and >45 (n = 17). All the patients underwent ovarian stimulation using a GnRH antagonist protocol and received a GnRH agonist trigger. The main factors used to measure outcome were oocyte yields, days of stimulation, gonadotrophin dosages, biochemical pregnancy, ongoing pregnancy and miscarriage rates. RESULTS: No differences between the study group and controls were reported in oocyte yields (17.5 versus 18.9) or days of stimulation (11.40 versus 9.82). The control group used significantly more gonadotrophin (2212 versus 1850 IU) than the study group. Clinical outcome was not affected by the CGG repeats on the FMR1 gene in oocyte donors. CONCLUSIONS: No negative effect was observed for intermediate-sized CGG repeats on ovarian stimulation and clinical outcome using a non-confounding model of oocyte donation. These results disagree with previous studies performed on infertility patients. Owing to the present study, fragile X genetic screening should not be considered for prediction of response to ovarian stimulation.

Circulation of zoonotic flaviviruses in wild passerine birds in Western Spain.

We explore the presence of zoonotic flaviviruses (West Nile virus (WNV) and Usutu virus (USUV)) neutralizing antibodies in rarely studied passerine bird species. We report, for the first time in Europe, WNV-specific antibodies in red avadavat and cetti's warbler, and USUV in yellow-crowned bishop. The evidence of WNV and USUV circulating in resident and migratory species has implications for both animal and public health. Future outbreaks in avian reservoir hosts may occur and passerines should be considered as priority target species in flavivirus surveillance programmes.

Operative start times and complications after liver transplantation.

The recent national focus on patient safety has led to a re-examination of the risks and benefits of nighttime surgery. In liver transplantation, the hypothetical risks of nighttime operation must be weighed against either the well-established risks of prolonging cold ischemia or the potential risks of strategies to manipulate operative start times. A retrospective review was conducted of 578 liver transplants performed at a single institution between 1995 and 2008 to determine whether the incidence of postoperative complications correlated with operative start times. We hypothesized that no correlation would be observed between complication rates and operative start times. No consistent trends in relative risk of postoperative wound, vascular, biliary, or other complications were observed when eight 3-h time strata were compared. When two 12-h time strata (night, 3 p.m.-3 a.m., and day, 3 a.m.-3 p.m.) were compared, complications were not significantly different, but nighttime operations were longer in duration, and were associated a twofold greater risk of early death compared to daytime operations (adjusted OR 2.9, 95% CI 1.16-7.00, p = 0.023), though long-term survival did not differ significantly between the subgroups. This observation warrants further evaluation and underscores the need to explore and identify institution-specific practices that ensure safe operations regardless of time of day.

The effect of dentine-bonding agents on substance P release in human dental pulp.

AIM: To quantify the effect of dentine-bonding agents on Substance P (SP) release in healthy human dental pulp tissue. METHODOLOGY: Forty pulp samples were obtained from healthy pre-molars where extraction was indicated for orthodontic reasons. In thirty of these pre-molars, a standardized Class V cavity preparation was performed, and teeth were divided equally into three groups: (i) Unetched-cavity control group: Class V cavities only; (ii) Experimental Group I: 'One-step' self-etch bonding agent was placed in the cavity; and (iii) Experimental Group II: 'Two-step' total-etch bonding agent was placed in the cavity. The remaining ten healthy pre-molars where extracted without treatment and served as an intact-teeth control group. SP was measured by radioimmunoassay. RESULTS: Greater SP release was found in the 'one-step' bonding agent group, followed by the 'two-step' bonding agent group and the unetched-cavity control group. The lower SP values were for the intact-teeth control group. anova showed statistically significant differences between groups (P = 0.0001). Tukey HSD post hoc tests showed statistically significant differences in SP release between the intact-teeth control group and the three other groups (P < 0.01) and between the unetched-cavity control group and the 'one-step' bonding agent group (P < 0.05). No significant difference was found between the 'two-step' bonding agent and the unetched-cavity control group. CONCLUSION: Dentine-bonding agents placed over Class V cavity preparations increased SP release. One-step dentine-bonding agents increased SP release most.

[Stevens-Johnson syndrome plus intrahepatic cholestasis caused by clindamycin or chlorpheniramine]. / Síndrome de Stevens Johnson más colestasis intrahepática inducido por clindamicina o clorfeniramina.

A 48-year-old woman was hospitalized with the diagnosis of hepatitis. She presented with symptoms of jaundice, headache, elevated bilirubin, and elevated hepatic enzymes. She related a recent episode of a bronchial infection that was treated during the previous eight days with paracetamol (500mg, 2 doses only), chlorpheniramine, betamethasone and clindamycin. After an initial clinical and laboratorial improvement, she began to complain of pruritus of the palms and soles. Thereafter, vesicles evolving to blisters developed and a deterioration of her general health ensued. Serologies for hepatitis A, B, and C viruses were negative. Intrahepatic cholestasis and Stevens Johnson Syndrome (SJS) were the final diagnosis. The association of the Stevens Johnson Syndrome and intrahepatic cholestasis simultaneously, related to adverse drug reactions, is very rare. The drugs reportedly involved are mainly antibiotics, such as ampicillin, vancomycin, amoxicillin/clavulinic acid and erythromycin. Other drugs involved are non-steroidal anti-inflamatory drugs, such as mefenamic acid, ibuprofen, and sulindac. The reactions can be minor or severe and can even cause death, an outcome that has been reported in patients of all races and ethnic groups, but appears to be more rare in patients of Latin origin. We present a discussion of this case and review the main characteristics of the Stevens Johnson Syndrome.

Analysis of the functional relevance of a putative regulatory SNP of PDCD1, PD1.3, associated with systemic lupus erythematosus.

This study aimed to test the functional effects of the PD1.3 single nucleotide polymorphism (SNP) (rs11568821), which were proposed based on its association to systemic lupus erythematosus (SLE) susceptibility and in electrophoretic mobility shift assays (EMSA) results. We analysed transcriptional effects of the PD1.3 locus by enhancer reporter assays. Results were against the hypothesis that the PD1.3 locus acts as enhancer in transcriptional regulation of PDCD1. In addition, they excluded a differential effect of the PD1.3 alleles. EMSA results confirmed that oligonucleotides with the PD1.3 G allele bind RUNX1 but not those with the A allele. However, binding to PD1.3 G oligonucleotides was much lower than binding to positive control oligonucleotides. Criss-cross experiments showed that this was due to flanking nucleotides in the PD1.3 sequence that negatively affect RUNX1 binding. These results cast doubts on the functional relevance of the PD1.3 SNP and, together with the lack of association in several studies, put into question its role as an SLE susceptibility factor. Investigation of other PDCD1 polymorphisms is needed to uncover the possible effect of this gene on SLE susceptibility.

Mesh plug and ureteral necrosis after kidney transplantation.

CASE REPORT: We present a 59-year-old male who underwent living related kidney transplantation. During the procedure a previously placed mesh plug was found to be adherent to structures in the vicinity of the area of implantation. After revascularization, neocystouretostomy was performed with the ureter over the spermatic cord in order to avoid excessive dissection through this adherent mass. Shortly thereafter the patient was diagnosed with a ureteral leak and returned to the operating room. The intraoperative findings included adhesion of the ureter to the mesh plug with necrosis of its distal portion. A redo neocystouretostomy was performed. The patient recovered uneventfully. CONCLUSION: This case report illustrates a unique cause of a urinary fistula related to mesh plug open inguinal hernia repair and demonstrates the importance of ensuring a safe distance between vital structures and any prosthetic device.

Body condition and protein supplementation positively affect periovulatory ovarian activity by non LH-mediated pathways in goats.

Effects of rumen undegradable intake protein (UIP) supplementation on ovarian activity and serum insulin, GH, and LH were evaluated in goats having low or high body condition (BC). Goats with either low BC (n=16, 28.7+/-0.8 kg BW, BC=2.1+/-0.3) or high BC (n=16, 38.4+/-0.8 kg, BC=3.2+/-0.3) received, during 40-days, one of the two protein supplementation levels: without UIP or with UIP (120 g goat(-1)d(-1)). Oestrus was synchronized with two i.m. doses of PGF(2alpha), and jugular blood samples were collected from 36 to 42 h after the second prostaglandin injection at 15 min intervals. Serum concentrations of insulin, LH, and GH were measured The number of preovulatory follicles and the number of corpora lutea (CL) were evaluated by transrectal ultrasonography at 1 and 4 days after the second prostaglandin dose, respectively. Does with higher BC had more CL than those in the lower condition group (2.8+/-0.2 versus 1.8+/-0.2, P<0.05). Similarly, goats receiving UIP supplementation had more follicles (2.6+/-0.2 versus 1.9+/-0.2, P<0.05) and tended to have more CL (2.6+/-0.2 versus 2.0+/-0.2, P=0.05) than does not receiving UIP. Neither BCS nor UIP supplementation affected serum GH or LH concentrations, pulsatility, or area under the curve. High BC does produced more insulin (1.92+/-0.17 versus 0.81+/-0.17 ng/mL, P<0.01 ng/mL) than lower BC goats; the same for UIP-supplemented (1.69+/-0.18 versus 1.04+/-0.18, P<0.05). Results suggest that the increased ovarian activity observed in both UIP-supplemented and higher BC goats was not the result of changes in LH or GH, suggesting effects at a local level, through changes in insulin in a non-GnRH-gonadotrophin dependent manner.

Comprehensive mitochondrial DNA analysis and IVF outcome.

STUDY QUESTION: Do mitochondrial DNA (mtDNA) copy number and heteroplasmy in human embryos affect the ongoing pregnancy rate? SUMMARY ANSWER: Our study suggests that mtDNA copy number above a specific threshold is associated with the ongoing pregnancy rate. WHAT IS KNOWN ALREADY: Mitochondria play a vital role in cell function. Recently, there has been increasing research on mtDNA as a biomarker of embryo implantation. Although reports showed that high levels of mtDNA in the blastocyst are associated with low implantation potential, other publications were unable to confirm this. Confounding factors may influence the mtDNA copy number in euploid embryos. On the other hand it has been speculated that both mtDNA heteroplasmy and copy number contribute to mitochondrial function. Next generation sequencing (NGS) allows us to study in depth mtDNA heteroplasmy and copy number simultaneously. STUDY DESIGN SIZE DURATION: A prospective non-selection study was performed. We included 159 blastocyst biopsies from 142 couples who attended our clinic for preimplantation genetic testing for aneuploidies (PGT-A), from January 2017 to December 2017. All embryos were biopsied on Day 5 or Day 6. The aneuploid testing was performed by NGS. All blastocysts were diagnosed as euploid non-mosaic and were transferred. The mtDNA analysis was performed once the embryo diagnosis was known. PARTICIPANTS/MATERIALS SETTING METHODS: Sequencing reads mapping to the mtDNA genome were extracted from indexed bam files to identify copy number and heteroplasmy. The relative measure of mtDNA copy number was calculated by dividing the mtDNA reads by the nuclear DNA value to normalize for technical variants and the number of cells collected at the biopsy. All the results were subjected to a mathematical correction factor according to the embryo genome. Heteroplasmy was assigned by MitoSeek. MAIN RESULTS AND THE ROLE OF CHANCE: The mean average copy number and SD of mtDNA per genome was 0.0016 ± 0.0012. Regarding heteroplasmy, 40 embryos were heteroplasmy carriers (26.32%). MtDNA variants were detected in coding and non-coding regions and the highest number of variants in an embryo was eight. With respect to IVF outcome for mtDNA copy number analysis, we set a threshold of 0.003 for the following analysis. The vast majority of the embryos were below the threshold (142/159, 89.31%) and 17 embryos were classified as having higher mtDNA levels. We showed a reduction in ongoing pregnancy rate associated with elevated mtDNA copy number (42.96% versus 17.65%, P < 0.05). This result was independent of maternal age and day of the biopsy: these factors were included as confounding factors because mtDNA copy number was negatively correlated with female age (25 -30 y: 0.0017 ± 0.0011, 30 -35 y: 0.0012 ± 0.0007, 35 -40 y: 0.0016 ± 0.0009, over 40 y: 0.0024 + 0.0017, P < 0.05). Embryos biopsied on Day 5 were more likely to have higher quantities of mtDNA compared with those biopsied on Day 6 (0.0017 versus 0.0009, P < 0.001). According to IVF outcome and heteroplasmy, a lower ongoing pregnancy rate was reported for embryos that carried more than two variants. However, this did not reach statistical significance when we compared embryos with a number of variants lower or higher than two (39.15 versus 20.0, P = 0.188). Finally, a clear positive association between the mtDNA variants and copy number was reported when we compare embryos with or without heteroplasmy (0.0013 ± 0.0009 versus 0.0025 ± 0.0014, P < 0.001) and among different numbers of variants (0:0.0013 ± 0.0009, 1-2:0.0023 ± 0.0012, >2:0.0043 ± 0.0014, P < 0.05). LIMITATIONS REASONS FOR CAUTION: A limitation may be the size of the sample and the high-throughput sequencing technology that might not have detected heteroplasmy levels below 2% which requires high sequence depth A clinical randomized trial comparing the clinical outcome after the transfer of embryos selected according to mtDNA levels or only by morphological evaluation will be necessary. More research into the impact of mtDNA heteroplasmy and copy number on IVF outcome is needed. WIDER IMPLICATIONS OF THE FINDINGS: Our results demonstrate that embryos with elevated mtDNA copy number have a lower chance of producing an ongoing pregnancy. MtDNA copy number is higher in older women and is dependent upon the number of cell divisions that preceded biopsy. Moreover, our data suggest that mitochondrial activity could be a balance between functional capacity and relative mtDNA copy number. STUDY FUNDING/COMPETING INTERESTS: There are no conflicts of interest or sources of funding to declare. TRIAL REGISTRATION NUMBER: Not applicable.

Effect of sex-sorting and cryopreservation on the post-thaw sperm quality of Iberian red deer spermatozoa.

This study investigated the effect of sex-sorting and cryopreservation on post-thaw characteristics and fertility of red deer (Cervus elaphus) sperm for the first time. Semen was collected by electroejaculation from 10 mature stags during the breeding season, and each ejaculate split into four experimental groups: Bulk sorted spermatozoa, sorted but not sexed (BSS); sorted high purity X-spermatozoa (XSS); sorted high purity Y-spermatozoa (YSS); and, control non-sorted spermatozoa (NS). Following, all samples were frozen over liquid nitrogen. Two straws per stag and sample type were analyzed immediately post-thaw and following a 2-h incubation period at 37 °C. Post-thaw total motility (TM) as assessed by CASA was not different (P < 0.05) among NS, BSS and YSS sperm. For XSS, post-thaw TM was lower (39%, P < 0.05) than that for NS (54%) or BSS (50%), but similar (P > 0.05) to that of YSS (47%) sperm. The percentage of apoptotic spermatozoa as assessed by PI/YO-PRO-1 and flow cytometry analysis, was higher (17%, P ≤ 0.05) for XSS sperm than NS (12%), BSS (13%) and YSS (14%) sperm. Following incubation there were no differences (P > 0.05) in TM or percent apoptosis among treatments. Post-thaw chromatin stability calculated as the DNA fragmentation index (%DFI) was similar among treatments; following incubation %DFI increased in all except YSS, which displayed the lowest value (P < 0.05). Artificial insemination of synchronized hinds yielded 44, 52 and 62% delivery rates for YSS, NS and standard frozen-thawed sperm, respectively (P < 0.05). Notably, 93 and 55% of fawns born were males for the YSS and NS spermatozoa, respectively (P < 0.05). In summary, Y-sorted sperm displayed acceptable post-thaw sperm evaluation parameters and the expected offspring sex ratio. More studies are needed to understand the source of sperm damage that may compromise the fertility of Y-sorted red deer sperm.

Dietary patterns and risk of nonfatal acute myocardial infarction in Costa Rican adults.

OBJECTIVE: To investigate the association between dietary patterns and risk of a first nonfatal acute myocardial infarction (MI) in Costa Rican adults. DESIGN: Population-based case-control study. SUBJECTS: A total of 496 incident MI cases and 518 population-based randomly selected controls matched to the cases by age (+/-5 years), gender, and county of residence. Subjects were interviewed with a validated food frequency questionnaire. Dietary patterns were identified by factor analysis. Odds ratios (OR) and 95% confidence intervals (CI) were obtained using multivariate conditional logistic regression adjusted for several recognized risk factors for MI. RESULTS: Two diet patterns were identified, 'vegetable' characterized by increased intake of vegetables and fruits, and 'staple', characterized by an increased use of palm oil for cooking, and intake of refined grains (mostly white rice and white bread), legumes, coffee, added sugar, and red meat. Compared to the lowest quintile of the staple diet pattern, the highest quintile was associated with an increased risk of MI (OR: 3.70, 95% CI: 2.30-5.97). Adjusting for potential confounders did not change the results (OR: 3.53, 95% CI: 1.98-6.31). Consistently, an increasing staple pattern score was associated with lower HDL cholesterol (P for trend <0.02) and alpha-linolenic acid in adipose tissue (P for trend <0.0001). The vegetable pattern was not associated with MI. CONCLUSIONS: The staple dietary pattern of Costa Rican adults is associated with low plasma HDL cholesterol, low alpha-linolenic acid in adipose tissue, and increased risk of MI.