Culture of human corpus cavernosum endothelium

Summary A method for culturing endothelial cells (HCC-EC) from surgical specimens of human corpus cavernosum has been developed. The approach involves selective endothelial outgrowth from explants and may be generally applicable to tissue whose endothelium is not amenable to isolation by routine mechanical or enzymatic methods. The tissue is minced into pieces which are placed onto gelatin-or fibronectin-coated tissue culture plastic, and grown in medium suitable for microvascular endothelial cell growth (Carson and Haudenschild, In Vitro 22:344–354, 1986). By Days 5 to 7 EC colonies are found. Within a day or two after the appearance of the EC colonies, a non-EC cell type appears and, if undisturbed, quickly overgrows the EC. An exploitable temporal separation between the emergence of EC and non-EC is obtained when both conditioned medium (from bovine aortic endothelium) and retinal extract are present during the outgrowth period. Explants are removed by pipetting at the first sign of the emergence of the non-EC cell type. Once isolated, HCC-EC do not require conditioned medium but do require either retinal extract or acidic fibroblast growth factor for survival and growth. Approximately 60% of the first passage cultures are at least 80% EC as judged by DiI-Ac-LDL labeling. One corpus (0.3×0.3×0.5 cm) usually produces 120 cm² of primary culture within 2 wk. These EC form contact-inhibited monolayers and stain positively for Factor VIII. They have a doubling time at 6th passage of 48 h and a plateau density of 5 to 7×10⁴ cells/cm². The availability of such cultures should facilitate the study of endothelium-mediated responses which play an important role in the erectile function of human penile corpus cavernosum. Supported by NIH R01 HL23567-09, DK-39080-01, DK40025-01, DK40487-01.     

Cave insects with sex‐reversed genitalia had their most recent common ancestor in West Gondwana (Psocodea: Prionoglarididae: Speleketorinae)

The divergence date and ancestral distributional area of the psocid subfamily Speleketorinae, which includes taxa with reversed genitalia (female penis and male vagina of Afrotrogla and Neotrogla, tribe Sensitibillini), were estimated. The most basal divergence of the subfamily (between the North American Speleketor and the tribe Sensitibillini) was estimated to have occurred according to the separation between the North American continent and Gondwana, ca. 175 Ma. The most basal divergence of Sensitibillini (between African Afrotrogla + Sensitibilla and Brazilian Neotrogla) was estimated to have occurred according to the split of West Gondwana (separation between the African and South American continents), ca. 127 Ma. The biome of the ancestral distributional area of Sensitibillini (inland of West Gondwana) is believed to be arid to semi‐arid, which might strengthen the reversed sexual selection and then facilitate the origin of preadaptive features related to the evolution of a female penis. All extant Sensitibillini species inhabit carbonatic caves, but geological evidence suggested independent shifts of these genera to the carbonatic cave habitat in the Tertiary/Quaternary.     

Detecting Concealed Knowledge Using a Novel Attentional Blink Paradigm

Traditional concealed information paradigms rely on the idea that stimuli that are meaningful to a person (critical items) will draw attentional resources disproportionately, relative to stimuli that are not (irrelevant items), generating detectable differences on a suitable dependent variable (behavioral, psychophysiological, or neural). Here, we introduce a behavioral paradigm that could be used to reveal concealed information by exploiting the link between concealed information and attentional processes more directly. This novel paradigm is based on the attentional blink phenomenon in which detection of a stimulus reduces detection accuracy rates of subsequent target stimuli within a 200–500 ms time window. We hypothesized that a well-known face used as a critical item could capture attentional resources automatically, making it harder to detect the occurrence of a subsequently presented target face. The results confirmed this hypothesis, and showed that concealed knowledge of a famous face could be detected in 9 out of 12 individuals by looking for a relative dip in target detection accuracy after the presentation of a critical item.     

Two intromittent organs in Zorotypus caudelli (Insecta,Zoraptera): the paradoxical coexistence of an extremely long tube and a large spermatophore

Very unusual genitalia of the species Zorotypus caudelli are described. It contains the unique configuration of two different intromittent organs, one of them strongly elongated. Hyper elongated genitalia are known in different groups of insects. Males have to accommodate these unwieldy structures in the limited spaces of the abdomen and manipulate them acutely during copulation. A crucial question is how do species with elongated genitalia cope with these requirements? To investigate this, we studied key features enabling storage, insertion, and withdrawal of the elongated genitalia. The co‐existence of an elongated narrow tube and a bulky spermatophore is a highly unusual and apparently paradoxical condition. However, we demonstrate that the tube is not involved in sperm transmission, whereas the large spermatophore is transferred to females by a membranous fold of the genitalia. The movement of the spermatophore is caused by haemolymph pressure, which likely also promotes the insertion of both intromittent organs. A comparison with the genital anatomy and reproductive mode in related groups suggests that the elongated tube and its accommodating pouch is a de novo structure, and that the ancestral sperm transport via spermatophore is a preadaptive condition for the acquisition of this unusual structure. © 2014 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 112 , 40–54.     

Why hide? Concealed sex in dominant Arabian babblers (Turdoides squamiceps) in the wild

Striking uniformity exists in humans' preference to conceal sexual activity from conspecifics' view. Yet, little is known about the selective pressures acting upon its evolution. To investigate this question, we studied the cooperatively breeding Arabian babbler (Turdoides squamiceps), which has been suggested being the only other species where dominant individuals conceal sex regularly. We examined whether birds indeed conceal sex and tested different hypotheses postulating that sex concealment functions to avoid predators, signal dominance status, or to avoid social interference. The results showed that the birds concealed sex in all observed cases of copulation, did not prefer to copulate under shelters and concealed mating solicitations from adult conspecifics. In addition, subordinates did not attack dominants who courted the respective female. Hence, none of the tested hypotheses explains these results satisfactorily. We postulate that dominant Arabian babblers conceal sex to maintain cooperation with those helpers they prevent from mating. Empirical desiderata for testing this ‘Cooperation-Maintenance’ hypothesis are discussed.     

Vascular endothelial growth factor induces IP-10 chemokine expression

We have previously shown that intracavernous injection of vascular endothelial growth factor (VEGF) improved the recovery of erectile function in an arteriogenic impotence rat mode. We wished to identify genes that are affected by VEGF treatment in the penis. Specifically we examined the induction of IP-10 chemokine. Male rats were subjected to pudendal arterial ligation or sham operation. They were then treated with intracavernous injection of 4 microg of VEGF in phosphate-buffered saline (PBS) or PBS alone. At 6 and 24 h after treatment, the erectile tissue was then harvested for RNA isolation. The isolated RNA was used for microarray and RT-PCR analyses. Cultured rat cavernous smooth muscle cells (CSMC) were treated with VEGF and then subjected to RT-PCR analysis. Cultured human CSMC were treated with VEGF and then subjected to ELISA analysis. Microarray analysis detected IP-10 as an abundantly induced message in 6-h VEGF-treated tissues. This was further confirmed by RT-PCR analysis. Using cultured rat CSMC, induction of IP-10 mRNA was detectable in 1 and 2 h, but not 24 h, VEGF-treated cells. Induction of IP-10 at the protein level was observed with cultured human CSMC. Secretion of IP-10 into culture medium peaked at 4 h after treatment of human CSMC with 10 ng/ml of VEGF. Optimal VEGF dosage for IP-10 induction was 50 ng/ml when assayed with cells that were treated for 8 h.     

Novelties of conception in insectivorous mammals (Lipotyphla), particularly shrews

In the order Lipotyphla (Insectivora), certain reproductive features differ quite distinctly from the eutherian norms, and are of interest with regard to the evolution of mammalian gamete function and perhaps for questions of lipotyphlan phylogeny. As seen in one or more members of five lipotyphlan families (shrews, moles, hedgehogs, golden moles, tenrecs), these features can involve the configuration of the male tract including the penis, the morphology of the sperm head, the anatomy of the oviduct and the patterns of sperm transport within it, the character of the cumulus oophorus, and the way in which fertilising spermatozoa interact with the eggs. However, the picture is by no means uniform within the order. Reproductive idiosyncrasies occur variously in the different lipotyphlan families, and appear consistently and strikingly in shrews--the group studied most extensively. Compared to the patterns in most Eutheria, the most interesting anomalies in soricids include (a) the regulation of sperm transport to the site of fertilisation by oviduct crypts, whose arrangement can vary even according to species, (b) a circumscribed matrix-free cumulus oophorus that appears essential for fertilisation as the inducer of the acrosome reaction, (c) barbs on the acrosome-reacted sperm head by which it may attach to the zona pellucida. With regard to the bearing such reproductive traits might have on lipotyphlan systematics, the African mouse shrew (Myosorex varius) displays a mix of traits that characterize either crocidurine or soricine shrews, consistent with the proposal that it belongs in a more primitive tribe, Myosoricinae, or subfamily, the Crocidosoricinae, from which the crocidurine and soricine lines probably evolved. Moreover, although elephant shrews are assigned now to a separate order (Macroscelidea), they display several of the unusual reproductive features seen in lipotyphlans, particularly in chrysochlorids and tenrecs. On the other hand, if used as a phylogenetic yardstick, none of the reproductive features described serves to define the Lipotyphla as classically constituted within one order, nor necessarily all the relationships suggested by recent sequencing studies of nuclear and mitochondrial genes.     

Countering Countermeasures in the Concealed Information Test Using Covert Respiration Measures

The effects of physical and mental countermeasures on the accuracy of the concealed information test (CIT) were examined in a mock crime experiment with 64 participants. To combat countermeasures, two covert respiration measures, hidden in the seat and back of the examination chair, were used in addition to the standard physiological measures (SCR, FPWL, RLL). Some guilty participants were trained to use either physical or mental countermeasures and apply them to distort the outcomes of the CIT. In the second phase of the experiment participants were detached from the standard polygraph devices and examined solely with the two covert measures. Results indicated that physical countermeasures lowered SCR accuracy but had a relatively small effect on the other standard measures. On the other hand, SCR was relatively resistant to mental countermeasures. Both covert measures were resistant to physical countermeasures in the polygraph phase. When the standard devices were removed, the covert seat measure was effective in the no countermeasure and in the mental countermeasure conditions but not when physical countermeasures were applied. The back measure was entirely ineffective.

Structural specialties,curiosities, and record‐breaking features of crustacean reproduction

Crustaceans are a morphologically, physiologically, and ecologically highly diverse animal group and correspondingly diverse are their reproductive characteristics. They have evolved structural specialties with respect to penis construction, sperm form, sperm storage, fertilization, and brood care. Unique in the animal kingdom are safety lines that safeguard hatching and first molting. Further curiosities are dwarf males in parasitic and sessile crustaceans and bacteria‐induced feminization and gigantism of crustacean hosts. Record‐breaking features are relative penis length, sperm size, clutch size, chromosome number, viability of dormant eggs, and fossil ages of penis, sperm, and brooded embryos. These examples from a single invertebrate subphylum and a single life history aspect illustrate that morphological solutions to functional requirements can be as spectacular as behavioral adaptations. They may provide valuable sources for comparative morphologists, ecologists, evolutionary biologists, and applied biologists to advance topical issues such as sperm competition, posthumous paternity, evolution of brood care, adaptation to freshwater, infectious feminization, sustainable male‐based fishery, maintenance of genetic diversity under conditions of limited mating opportunity, and long‐term impact of pollution on genotype and phenotype. J. Morphol. 277:1399–1422, 2016. © 2016 Wiley Periodicals, Inc.