One of the most recognizable cases of preimplantation genetic diagnosis (PGD) is X-linked diseases. Diagnosis of fetal sex is essential for couples who are known to be at risk of some X-linked disorders. The objective of this study was to discriminate between female (XX) and male (XY) embryos by detecting sex chromosomes-specific sequences in spent culture medium and comparing these results to PGD/CGH array results. It may open new window for the development of a non-invasive PGD method. 120 Embryo’s spent media from Day 3 and Day 5 embryos were collected. Modified phenol-chloroform solution was used for DNA extraction from spent media. Sex determination was performed using SRY, TSPY and AMELOGENIN evaluation through quantitative polymerase chain reaction (q-PCR) method. IBM SPSS and MedCalc were used for statistical analyses to compare sex determination of embryos by spent medium with PGD/CGH array results. Culture time was demonstrated to increase the DNA amount among day 5 embryos culture medium samples. Non-invasive PGD by means of spent culture medium gave a sensitivity, specificity, positive predictive value and negative predictive value of 100% for sex determination. Results of sex determination using spent medium by q-PCR were consistent with the results of PGD/CGH array. Improvements in cell-free DNA extraction and PCR amplification procedures provide us an effective method to perform a PGD test without biopsy in the future, especially about X-linked diseases.

Waterhemp [Amaranthus tuberculatus (Moq.) J. D. Sauer] and Palmer amaranth (Amaranthus palmeri S. Watson) are troublesome weeds of row-crop production in the United States. Their dioecious reproductive systems ensure outcrossing, facilitating rapid evolution and distribution of resistances to multiple herbicides. Little is known, however, about the genetic basis of dioecy in Amaranthus species. In this work, we use restriction site–associated DNA sequencing (RAD-Seq) to investigate the genetic basis of sex determination in A. tuberculatus and A. palmeri. For each species, approximately 200 plants of each sex were sampled and used to create RAD-Seq libraries. The resulting libraries were separately bar-coded and then pooled for sequencing with the Illumina platform, yielding millions of 64-bp reads. These reads were analyzed to identify sex-specific and sex-biased sequences. We identified 345 male-specific sequences from the A. palmeri data set and 2,754 male-specific sequences in A. tuberculatus. An unexpected 723 female-specific sequences were identified in a subset of the A. tuberculatus females; subsequent research, however, indicated female specificity of these markers was limited to the population from which they were identified. Primer sets designed to specifically amplify male-specific sequences were tested for accuracy on multiple, geographically distinct populations of A. tuberculatus and A. palmeri, as well as other Amaranthus species. Two primer sets for A. palmeri and four primer sets for A. tuberculatus were each able to distinguish between male and female plants with at least 95% accuracy. In the near term, sex-specific markers will be useful to the A. tuberculatus and A. palmeri research communities (e.g., to predict sex for crossing experiments). In the long-term, this research will provide the foundational tools for detailed investigations into the molecular biology and evolution of dioecy in weedy Amaranthus species.

Several populations of different Amaranthus species have been reported resistant to single or multiple herbicides. Interspecific hybridization among amaranths is hypothesized to contribute to the evolution of herbicide resistance. Although other studies have shown the occurrence of interspecific Amaranthus hybrids, little has been done to establish the likelihood of hybridization under field conditions. The main objective of this study was to determine potential field frequencies of hybridization between tall waterhemp females and smooth pigweed. Field hybridization plots were established during each of two growing seasons. Individuals of the two species were transplanted to field plots and arranged at varying distances from each other. Hybrid progeny were detected using the acetolactate synthase (ALS) gene as a marker. Smooth pigweed parents were homozygous for a herbicide-resistance ALS allele, whereas maternal tall waterhemps were homozygous for a herbicide-sensitive ALS form. Heterozygous interspecific progeny were thus detected by means of herbicide selection. Molecular and cytogenetic tools were used to verify the validity of the data obtained. Averaged among female waterhemp plants and across the two field seasons, hybridization occurred at a frequency of 33%. A single tall waterhemp plant was capable of producing more than 200,000 hybrids, suggesting little if any gametic incompatibility. All flowering hybrids obtained from tall waterhemp females were of dioecious condition and female sex. Observed sexual segregation was consistent with previously postulated chromosomal XY-type system in tall waterhemp sex determination, where males are the heterogametic sex.

Classical and reverse genetics remain invaluable tools for the scientific investigation of model organisms. Genetic analysis of endoparasites is generally difficult because the sexual adults required for crossing and other manipulations are usually hidden within their host. Strongyloides spp. and Parastrongyloides spp. are notable exceptions to this and their free-living adults offer unique opportunities to manipulate these parasites experimentally. Here I review the modes of inheritance in the two generations of Strongyloides/Parastrongyloides and I discuss the opportunities and the limitations of the currently available methodology for the genetic analysis of these two genera.

Parasitic lifestyles evolved many times independently. Just within the phylum Nematoda animal parasitism must have arisen at least four times. Switching to a parasitic lifestyle is expected to lead to changes in various life history traits including reproductive strategies. Parasitic nematode worms of the genus Strongyloides represent an interesting example to study these processes because they are still capable of forming facultative free-living generations in between parasitic ones. The parasitic generation consists of females only, which reproduce parthenogenetically. The sex in the progeny of the parasitic worms is determined by environmental cues, which control a, presumably ancestral, XX/XO chromosomal sex determining system. In some species the X chromosome is fused with an autosome and one copy of the X-derived sequences is removed by sex-specific chromatin diminution in males. Here I propose a hypothesis for how today's Strongyloides sp. might have evolved from a sexual free-living ancestor through dauer larvae forming free-living and facultative parasitic intermediate stages.

To develop a philosophical framework to address the specification of sex/gender in humans, I will begin by summarizing what we know about the biology of sex determination in human beings. Basically, sex/gender is specified by several networks of genes anchored in the genetic interaction between the two genes, Sry and Sox9. Next, I will propose that this biological mechanism is best understood within a philosophical anthropology that embraces insights taken from systems biology to articulate a hylomorphism that explains the integrity, dynamism, and teleology of the human organism. The systems perspective described here represents one attempt to reformulate the received philosophical framework of classical Aristotelian-Thomistic hylomorphism so that it incorporates the insights of modern biology. Finally, I will use the systems perspective and key principles articulated by St. Thomas Aquinas in his philosophy of nature to identify criteria that could be used to specify the sex/gender of a particular human being: The most certain criterion for maleness would be the capacity to produce sperm while the most certain criterion for femaleness would be the complementary capacity to produce eggs. Deviations from this criterion would decrease the certitude of our judgment regarding the sex/gender of the individual.

The parasitic roundworms Strongyloides stercoralis (in man) and Strongyloides ratti (in rats) employ environmentally controlled XX/XO sex determination with a pair of X chromosomes and two pairs of autosomes. Strongyloides papillosus (in sheep) has only two pairs of chromosomes, one of which combines the genetic material homologous to the S. ratti chromosomes X and I. This species creates males through the elimination of one copy of the portion related to the X chromosome (chromatin diminution). It is not clear which one of these two sex-determining mechanisms is ancestral. We demonstrate that Strongyloides vituli (in cattle) has two pairs of chromosomes like its very close relative S. papillosus whereas Parastrongyloides trichosuri, a closely related out-group to Strongyloides spp. in Australian brushtail possums, has three chromosome pairs and employs XX/XO sex determination. The X chromosome of P. trichosuri is homologous to the X chromosome of S. ratti. Our data strongly suggest that the last common ancestor of Strongyloides spp. and Parastrongyloides spp. had two pairs of autosomes along with two or one X chromosome in females and males, respectively. The situation with two pairs of chromosomes is likely derived and occurred through the fusion of the X chromosome with an autosome.

Sex ratios in farmed European sea bass are highly biased towards males (75 to 95%), which is problematic for aquaculture. In this mini-review, we re-analyse fisheries literature data about sex ratios in wild sea bass from 13 population samples, representing altogether 4889 individuals covering the major part of the distribution range of the species. We find that as a whole, the sex ratio of wild populations is biased towards females (59.4% females, p < 0.001), but that the sex ratio of the younger fish (<30 cm total length) is balanced (52.0% females, p = 0.15), while the sex ratio of the older fish is heavily biased towards females (69.5% females, p < 0.01). Possible causes of these differences (differential longevity, biased sampling) are discussed. When age-group sex ratios are available (three population samples out of 13), significant variation between age groups appears, part of which is most likely of environmental origin. This study shows that the excess of males in culture is not a characteristic of the species, but rather a consequence of the environments used in culture, interacting with a complex system where both environmental and genetic influences govern sex determination in sea bass.