HSP47 expression in the hamster Sertoli cell: An immunohistochemical study.

HSP47, a chaperone whose main function is the maturation of collagen molecules, is considered a marker of fibrotic diseases. Increased collagen synthesis in the testis has been associated with various pathologies leading to testicular seminiferous tubule regression. Our aim was to study whether HSP47 is expressed in hamster Sertoli cells both in the adult and in two physiological situations of seminiferous tubule atrophy: irreversible testicular ageing and testicular regression due to short (reversible) photoperiod. Eighteen animals were divided as follows: a group of 6 young animals aged 6 months, a group of 6 animals aged 24 months, which were exposed to a long photoperiod, and a final group of 6 young animals subjected to a short photoperiod. Testicular samples were fixed in methacarn and an immunohistochemical technique was used to detect HSP47. A semiquantitative study of the expression of this protein was performed between tubular sections of aged animals with complete spermatogenesis and arrested spermatogenesis with tubular sections with arrest spermatogenesis of photoinhibited testes. Sertoli cells were positive for HSP47, the intensity being higher in tubular sections with arrested spermatogenesis in both aged and photoinhibited animals. Sertoli cells were positive for HSP47, the intensity being greater in tubular sections with arrested spermatogenesis in both aged and photoinhibited animals. Semiquantitative analysis corroborated this observation in the sense that the expression of this protein differed according to the functional state of the seminiferous tubules. Thus, the intensity index of immunoreactivity was significantly higher in tubular sections with arrested spermatogenesis in aged animals compared with regressed animals, and in the latter compared with those whose tubular sections showed complete spermatogenesis. In conclusion, HSP47 expression in Sertoli cells was found for the first time in mammals. Moreover, increased expression seemed to be related to the degree of atrophy of the seminiferous epithelium and to the reversible or non-reversible physiological state of the seminiferous epithelium.

Extracellular vesicles would be involved in the release and delivery of seminal TGF-ß isoforms in pigs.

Introduction: Pig seminal plasma (SP) is rich in active forms of all three isoforms (1-3) of transforming growth factor ß (TGF-ß), a chemokine modulatory of the immune environment in the female genital tract once semen is delivered during mating or artificial insemination (AI). The present study aimed to examine how TGF-ßs are secreted by the epithelium of the male reproductive tract and how they are transported in semen, emphasizing the interplay with seminal extracellular vesicles (sEVs). Methods: Source of TGF-ßs was examined by immunohistochemistry in testis, epididymis, and accessory sex glands, by immunocytochemistry in ejaculated spermatozoa, and by Luminex xMAP® technology in SP and sEVs retrieved from healthy, fertile male pigs used as breeders in AI programs. Results: All three TGF-ß isoforms were expressed in all reproductive tissues explored and would be released into ductal lumen either in soluble form or associated with sEVs. Ejaculated spermatozoa expressed all three TGF-ß isoforms, both inside and outside, probably the outer one associated with membrane-bound sEVs. The results confirmed that pig SP contains all three TGF-ß isoforms and demonstrated that a substantial portion of them is associated with sEVs. Discussion: Seminal EVs would be involved in the cellular secretion of the active forms of seminal TGF-ß isoforms and in their safe transport from the male to the female reproductive tract.

Cellular Modifications in Spermatogenesis during Seasonal Testicular Regression: An Update Review in Mammals.

Testicular regression occurs during the non-breeding season in many mammals. This affects spermatogenesis, resulting in decreased or arrested activity. Both lead to a decrease or cessation in sperm production. In recent years, the cellular mechanisms that lead to infertility in males in non-reproductive periods have been studied in very different species of mammals. At the start of the present century, the main mechanism involved was considered as an increase in the apoptotic activity of germ cells during the regression period. The loss of spermatogonia and spermatocytes causes not only a decrease in spermatogenesis, but an arrest of the seminiferous epithelium activity at the end of regression. Recently, in some mammal species, it was found that apoptosis is the usual mechanism involved in epithelium activity arrest, although it is firstly atrophied by massive desquamation of the germ cells that are released from their binding with the Sertoli cells, and which are shed into the lumen of the seminiferous tubule. In other species, it has been shown that not only germ cell apoptosis, but also Sertoli cell apoptosis, including decreased proliferative activity, spermatophagy or autophagy, are involved in testicular regression. Furthermore, the most recent studies indicate that there are multiple patterns of seminiferous epithelium regression in seasonally breeding animals, which may not only be used by different species, but also by the same ones to reproduce in the best conditions, ensuring their survival. In conclusion, at this time, it is not possible to consider the existence of a paradigmatic cellular mechanism in the involution of the seminiferous epithelium applicable to all male mammals with seasonal reproduction, rather the existence of several mechanisms which participate to a greater or lesser extent in each of the species that have been studied to date.

[The application of technoscience to man: ethical discernment in relation to the transhumanist-posthumanist proposal]. / La aplicación de la tecnociencia al hombre: discernimiento ético en relación con la propuesta transhumanista-posthumanista.

Transhumanism-posthumanism is a current of thought that appears closely linked to the development of technoscience and its application to man. At the same time that this current must be subject to criticism, an anthropological and ethical paradigm must be illuminated that allows accepting or not the new technoscientific advances, making a discernment between them. Such discernment should lead us to weigh the goodness of these advances, rejecting only those that represent a degradation of the human being, and accepting those that help man to be more fully man. To do this, the article proposes starting the discernment from an ethical principle such as respect for the integrity of man. Together with him, it is necessary to act with caution regarding human health, considered in relation to his psychosomatic unity. It will also be necessary to avoid deriving the ethics of the advances from the same technoscience. Finally, discernment requires, ultimately, starting from an idea about what man is, proposing the need to do so from a dual rather than dualistic conception of the human person. Based on all of the above, various ethical criteria are indicated in the work that complete the principle of respect for human integrity indicated above: respect and promote human life in all its dimensions, use of technology at the service of human beings in a controlled manner and that report social benefit or value by each technique, not only from a therapeutic perspective, but also from the improvement of the human psychosomatic unit. In conclusion, it is necessary to recognize in man the uniqueness of him as he is a bodily being who knows and loves in freedom, whose ends are not limited to material or sensible things, but which are only achievable in and from his own material condition. Consequently, any techno-scientific intervention that substantially alters his body condition is inhuman, not instead when it repairs or enhances -without abolishing them- his own qualities.

Proliferation, apoptosis, and number of Sertoli cells in the Syrian hamster during recrudescence after exposure to short photoperiod†‡.

The Sertoli cell (Sc) has been described as a quiescent cell once the animal has reached sexual maturity. Syrian hamster is an animal that displays testicular regression due to short photoperiod, during which process germ cells and Sc are removed through apoptosis. The aim of this work was to investigate histochemically whether the spontaneous testicular recrudescence processes after exposure to a short photoperiod lead to an increase in Sc proliferative activity in order to restore the normal population. Three spontaneous recrudescence groups were established: initial (IR), advanced (AR), and total (TR) recrudescence, which were compared with animal undergoing the regression process (mild: MRg, strong: SRg, and total: TRg) and animals in long photoperiod (Controls). Histological sections were submitted to histochemical techniques for detecting apoptotic and proliferative Sc with bright-field and fluorescence microscopy. For each group, the proliferative Sc index (PScI) and apoptotic Sc index (AScI), and the total number of Sc were obtained. The results revealed the existence of Vimentin+/TUNEL+ as well as Vimentin+/PCNA+ cells. The PScI was significantly higher in TRg and IR than in the other groups. The AScI was only significantly higher in MRg and SRg with respect to the other groups. The total number of Sc increased among TRg, IR, and AR, reaching values similar to those of the Controls. In conclusion, the increase in Sc proliferation from final regression and recrudescence, accompanied by a similar rate of apoptosis to the Control group, is the cause of the restoration of the Sc population during spontaneous recrudescence.

[Ethical character and prudence: analysis of the human act in clinical-ethical decisions]. / Carácter ético y prudencia: analisis del acto humano en las decisiones clínico-éticas.

First, the mutual interiority that exists in every human action between its ethical and technical dimension is analyzed, more specifically in relation to health practice. It is established that by means of technical or ethical habits, or applying certain deliberative guidelines, alone it is not possible to discern between the goodness or not of a certain action. For this to be possible, it is necessary to adopt a personal perspective, both as regards oneself and when judging the actions of others. Secondly, then, the virtue of prudence is essential to choose the most best path in a specific context. This ethical habit is greatly influenced by the overall ethical character of the person and by their deliberative capacity and knowledge of ethical principles. With regard to the second, it is necessary to follow the usual ways by which the person makes decisions and considers a certain action as good or bad. Therefore, it cannot be pretend using technical decision-making methods health professionals will deliberate and make ethical decisions. In conclusion, it is necessary that the prudence of professionals be based both on the personal ethical character and on an analysis of ethical cases in consonance with the natural way in which human reason acts.

Lectin-binding pattern of glycoconjugates during spontaneous testicular recrudescence in Syrian hamster (Mesocricetus auratus) after exposure to short photoperiod.

Lectin histochemistry was used to characterise glycoconjugates and cellular apoptosis in the seminiferous epithelium and interstitium of hamster testis during spontaneous recrudescence. An increase in the LTA lectin affinity was observed in spermatids in the Golgi phase. An increase in labelling of PNA and Con-A lectin in acrosome of spermatids (acrosome phase) as well as increased labelling with Con-A in spermatids (cap phase) was observed. Spermatocytes showed decreased affinity with PNA and AAA lectins and an increase in positivity for LTA and GNA lectins. Spermatogonia showed a slight decrease in positivity to WGA and an increase in labelling with Con-A and a decreased affinity for the AAA lectin. At the end of recrudescence, all these germinal cells showed a similar pattern to the control. The Sertoli cells showed a gradual decrease in labelling with the GNA lectin and the Leydig cells an increase in labelling with Con-A and GNA. Particularly unusual was the observation of apoptotic spermatocytes and spermatids positive for PNA, GNA, AAA and Con-A, together with spermatocytes positive to LTA. In conclusion, the normal lectin pattern is recovered during testis recrudescence and germ cell apoptotic activity is low, as is observed by specific lectins for germ cells in apoptosis.

Testicular histomorphometry and the proliferative and apoptotic activities of the seminiferous epithelium in Syrian hamster during spontaneous recrudescence after exposure to short photoperiod.

Syrian hamsters are photoperiodic rodents in which reproduction, including testicular function, is stimulated by long photoperiod exposure and curtailed by exposure to a short photoperiod. The objectives of this study were to characterize the testis histomorphometrically and to determine the role of the proliferation and apoptosis phenomena in the recovery of the seminiferous epithelium during spontaneous recrudescence after exposure to short photoperiod. The study was performed using conventional light microscopy, proliferating cell nuclear antigen and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP in situ nick end labelling staining, image analysis software, and transmission electron microscopy in three recrudescence groups: initial recrudescence (IR), advanced recrudescence (AR) and total recrudescence (TR). The results morphometrically pointed to the gradual recovery of the testicular and tubular volumes, as well as of the seminiferous epithelium. Among the IR and AR groups, the increase in testicular and tubular volumes was accompanied by an increase in tubular diameter and length, with an increase in interstitial volume. From AR to TR, there was an increase in the tubular and total volumes, but, in this case, with a gradual increase in tubular diameter. Recovery of the seminiferous epithelium was accompanied by changes in apoptosis and proliferation activities. The first decreased halfway through the process, and the second remained higher than the control levels throughout the recrudescence stage. Alterations in the spermatozoa were ultrastructurally observed, which indicated that spermiogenesis was not yet completely normal. In conclusion, spontaneous testicular recrudescence in Syrian hamster comprises two histomorphometrical phases: the first related to an increase in tubular length and diameter and interstitial volume and the second depending principally on the gradual increase in tubular diameter. The restoration of the seminiferous epithelium is due to apoptosis reaching normal values in the AR group accompanied by higher proliferative activity than that observed in the Control group.

Identification of Proliferative and Apoptotic Sertoli Cells Using Fluorescence and Confocal Microscopy.

Sertoli cells, the testicular somatic cells of the seminiferous epithelium, are vital for the survival of the epithelium. They undergo proliferation and apoptosis during fetal, neonatal, and prepubertal development. Apoptosis is increased in certain situations such as exposure to many substances, for example, toxics, or short photoperiod in the non-breeding season of some mammals. Therefore, it has always been considered that Sertoli cells that reach adulthood are quiescent cells, that is to say, nonproliferative, do not die, are terminally differentiated, and whose numbers remain constant. Recently, a degree of both proliferation and apoptosis has been observed in normal adult conditions, suggesting that consideration of this cell as quiescent may be subject to change. All this make it necessary to use histochemical techniques to demonstrate whether Sertoli cells are undergoing proliferation or apoptosis in histological sections and to allow the qualitative and quantitative study of these. In this chapter, we present two double-staining techniques that can be used for identifying Sertoli cells in proliferation or apoptosis by fluorescence microscopy. In both, the Sertoli cells are identified by an immunohistochemistry for vimentin followed by an immunohistochemistry for PCNA or a TUNEL histochemistry.

[Use of the Term ″Pre-Embryo″ In the Biomedical Literature From Its Origin to the Present]. / Uso del Término ″Preembrión″ en la Literatura Biomédica Desde Su Origen Hasta la Actualidad.

In this article, we present a review about the use of the term ″preembryo″ in the scientific literature as well as Bioethics from its origin, back in June 1979, to the end of the year 2014. We analyze their evolution along the time comparing with other terms commonly used in embryology. Also, we show the relevant journals in which this term appears, (offering the impact index within their specific field), and the authors who use it the most. The term ″preembryo″ arose in the scientific literature in 1979, remaining for about 6 years without reappearing in the scientific literature. Then, after an increase of articles, that covers the 90's decade, began to decrease its use without having disappeared altogether. Our study also shows that the use of the word ″preembryo″ has not increased with the passage of time; on the contrary, it is becoming less used in the biomedical literature. This does not occur with other terms that name the embryo before implantation, which have not ceased to increase during these years, both referring to the human species and other animal species. Finally, the term ″pre-embryo″ appears very little in journals related to the reproduction biology area, but it does so in obstetrics and gynecology, where many articles on assisted reproduction are published. Our findings suggest that, instead of replacing classical embryological terms, the word ″preembryo″ does not seem to affect the use of them. Likewise, this word has a hypertrophied use in humans without an apparent reason, which would support the artificial nature of this term. Finally, the term ″pre-embryo″ appears very little in journals of reproduction biology area, but it does so in obstetrics and gynecology, where many articles on assisted reproduction are published. In conclusion, instead of substituting classical embryological terms, the word ″pre-embryo″ does not seem to affect the use of them and in the current discussions about the human condition of the embryo this term is no longer used.

The Use of Lectin Histochemistry for Detecting Apoptotic Cells in the Seminiferous Epithelium.

Lectin histochemistry is commonly used to characterize the pattern of glycoconjugates in cells and tissues. Recent studies show that alterations in these glycoconjugates are associated with the entry of cells into apoptosis. A widely used technique for the detection of apoptotic cell death is TUNEL. In this chapter, we study the sensitivity of both techniques to identify apoptotic cells in the testis of photo-inhibited Syrian hamster.

[Informed Consent in the Humanization of the Cesarean: A Preliminary Study]. / Consentimiento Informado en la Humanización de la Cesárea: Un Estudio Preliminar.

The informed consent (IC) is a process based on dialogue between the professional and the patient in which he freely decides on possible interventions in their health. This is applicable to caesarean delivery and if it meets a number of conditions will help to improve the process of ″humanization″ of birth. The overall objective of this study is to analyze preliminarily in several hospitals in the Region of Murcia the IC in caesarean delivery. To this end, we have revised the documents of IC and we studied who, where, when and how the IC process is done. The results show that all hospitals are based on the same document, and although the documents take into account all the elements of a IC, do not indicate the date of their design or subsequent revisions. It does not contemplate the risks and complications that caesarean section can have on the newborn, mother, and mother-child relationship later. It is noted that the document of IC normally is delivers by gynecologist in the consultation, when intervention is programmed, although it are sometimes nurses, who after admission to the hospital give it to sign the patient. In urgent caesarean sections, there are some hospitals that in life-threatening situation, do not offer the document of IC to women. In others, it is offered hastily by the gynecologist or midwife. In conclusion, the IC is a process which used correctly, favors the relationship between women and health professionals in the intervention of cesarean section. Although this process and the documents of IC examined in our study, have presented many positive aspects, the humanization of caesarean could be increased improving with the preparation and updating of these documents and coordinating the various professionals.