Loss of hamster Leydig cells during regression after exposure to a short photoperiod.

The aim of the present study was to evaluate the changes that occur in hamster Leydig cells during regression. Animals were divided into control, mild regression (MR), strong regression (SR) and total regression (TR) groups. Leydig cells were characterised by light and electron microscopy. Terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labelling (TUNEL) and proliferating cell nuclear antigen (PCNA) antibodies were used to detect apoptosis and proliferation respectively. Three types of Leydig cells (A, B and C) could be differentiated. Type A cells were small in size compared with Leydig cells from animals exposed to a long photoperiod, which was a result of a decreased cytoplasm and nucleus. Type B cells were even smaller than Type A cells in regression groups. Type C exhibited cytoplasm vacuolisation. The percentage of Type C cells from the control group was much lower than in the MR, SR and TR groups. (P<0.05). In the SR and TR groups, there was a significant decrease in the percentage of Type B cells compared with the control and MR groups (P<0.05). The total number of Leydig cells decreased during testicular regression (P<0.05). The total number of Type A and B cells was significantly lower in the MR, SR and TR groups compared with the control group (P<0.05). There were no significant differences in the proliferation and apoptosis index in the groups studied. The findings of the present study indicate that there are three types of Leydig cells (A, B and C) in all hamsters studied and that regression causes an increase in the number of Type C cells, so that the reduction in the number Leydig cells during the phases of regression studied must be the result of necrosis and/or necroptosis.

Cellular changes in the hamster testicular interstitium with ageing and after exposure to short photoperiod.

The aim of this study was to evaluate the cellular changes that occur in the hamster testicular interstitium in two very different physiological situations involving testicular involution: ageing and exposure to a short photoperiod. The animals were divided into an 'age group' with three subgroups - young, adult and old animals - and a 'regressed group' with animals subjected to a short photoperiod. The testicular interstitium was characterised by light and electron microscopy. Interstitial cells were studied histochemically with regard to their proliferation, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP in situ nick end labelling (TUNEL+) and testosterone synthetic activity. We identified two types of Leydig cell: Type A cells showed a normal morphology, while Type B cells appeared necrotic. With ageing, pericyte proliferation decreased but there was no variation in the index of TUNEL-positive Leydig cells. In the regressed group, pericyte proliferation was greater and TUNEL-positive cells were not observed in the interstitium. The testicular interstitium suffered few ultrastructural changes during ageing and necrotic Leydig cells were observed. In contrast, an ultrastructural involution of Leydig cells with no necrosis was observed in the regressed group. In conclusion, the testicular interstitium of Mesocricetus auratus showed different cellular changes in the two groups (age and regressed), probably due to the irreversible nature of ageing and the reversible character of changes induced by short photoperiod.

Changes in Testicular Interstitial Connective Tissue of Hamsters (Mesocricetus auratus) During Ageing and After Exposure to Short Photoperiod.

The testicular interstitium of Syrian hamster (Mesocricetus auratus) was studied during ageing and in testicular regression after exposure to a short photoperiod, in relation to the interstitial cells and their connective tissue. This tissue was assessed histochemically using Masson's trichrome technique and the expression of Heat Shock Protein 47 (HSP-47) and collagen IV (α5) was assessed in Leydig cells. Finally, an ultrastructural analysis of some cells of the testicular interstitium was made. Leydig cells were positive for HSP-47 and collagen IV (α5). Ageing did not change the parameters studied while the short photoperiod altered the synthetic activity of Leydig cells. The positivity index of these cells for HSP-47 was significantly higher in the regressed testis, but was lower for collagen IV (α5). During ageing no change were observed. Ultrastructural Leydig cells showed a discontinuous basal lamina that did not change during ageing. The basal lamina was not identified in Leydig cells regressed by exposure to a short photoperiod. In conclusion; the intertubular connective tissue suffers little change with age. By contrast, in the testis regressed after exposure to a short photoperiod the studied parameters related to the intertubular connective tissue were altered. These changes are probably related with the low synthetic activity of regressed Leydig cell.

Influence of histological degree of seminiferous tubular degeneration and stage of seminiferous cycle on the proliferation of spermatogonia in aged Syrian hamster (Mesocricetus auratus).

The ageing testis is associated with germ loss in the seminiferous epithelium and a decrease in spermatogonia proliferation. In this work, we study whether the stages of the seminiferous epithelium cycle and/or the degree of histological tubular degeneration resulting from ageing is related with this decrease in spermatogonia proliferation. Eleven hamsters were used, five aged 6 months and six aged 24 months. In both groups, the proliferative activity was studied by BrdU immunostaining. The number of BrdU-positive and BrdU-negative cells was measured, providing the overall proliferation index in adult and aged testes. The mean number of BrdU-positive cells was also determined for each degree of histological degeneration of seminiferous epithelium, and a spermatogonia proliferation index was obtained for each stage of the seminiferous cycle. Ageing caused an overall decrease in the BrdU-positive cell percentage and a decrease in the number of BrdU-positive cells in the tubular sections with hypospermatogenesis, the sloughing of germ cells and maturation arrest, these changes being similar in both young and old animals. The spermatogonia proliferation index was only seen to be significantly lower in ageing hamster in stages VII-VIII of the seminiferous epithelium cycle. In conclusion, the overall decrease in proliferation observed in aged seminiferous epithelium is correlated with an increase in the number of degenerated sections of the seminiferous tubules, and this decrease is a phenomenon which occurs in specific stages of the seminiferous cycle.

Lectin histochemistry as a tool to identify apoptotic cells in the seminiferous epithelium of Syrian hamster (Mesocricetus auratus) subjected to short photoperiod.

Lectins have been widely used to study the pattern of cellular glycoconjugates in numerous species. In the process of cellular apoptosis, it has been observed that changes occur in the membrane sugar sequences of these apoptotic cells. The aim of our work was to identify which lectins, out of an extensive battery of the same (PNA, SBA, HPA, LTA, Con-A, UEA-I, WGA, DBA, MAA, GNA, AAA, SNA), show affinity for germinal cells in apoptosis, at what stage of cell death they do so and in which germinal cell types they can be detected. For this, we studied testis sections during testicular regression in Syrian hamster (Mesocricetus auratus) subjected to short photoperiod. Several lectins showed an affinity for the glycoconjugate residues of germ cells in apoptosis: Gal ß1,3-GalNAcα1, α-d-mannose, N-acetylgalactosamine and l-fucose. Furthermore, lectin specificity was observed for some specific germinal cells and in certain stages of apoptosis. It was also observed that one of these lectins (PNA) showed affinity for Sertoli cells undergoing apoptosis. Therefore, we conclude that the use of lectin histochemistry could be a very useful tool for studying apoptosis in the seminiferous epithelium because of the specificity shown towards germinal cells in pathological or experimentally induced epithelial depletion models.

Corrosion behaviour and biocompatibility of a novel Ni-free intermetallic coating growth on austenitic steel by hot dipping in an Al-12.6%Si alloy.

Commercial 316 LVM austenitic stainless steel samples have been coated by immersion in a bath of molten Al-12.6%Si alloy for 120 s. The coating consists of the Al(12)(Fe,Cr)(3)Si(2) intermetallic. In vitro corrosion behaviour has been evaluated in the Ringer's solution by means of potentiodynamic curves and electrochemical impedance spectroscopy. The results reveal that the coated specimens exhibit lower susceptibility to localised corrosion with respect to the substrate. XPS analysis suggests that the ennoblement of the pitting potential is due to the formation of a chromium oxyhydroxide containing passive layer. The intermetallic coating shows a good biocompatibility, as demonstrated by culturing human mesenchymal stem cells isolated from bone marrow which attached, grew and differentiated to the osteoblastic lineage to a similar extent on coated and bare steels. In summary, this study proposes a method that generates Ni-free coatings of the stainless steel with useful properties for biomedical applications.

Proliferation and apoptosis in aged and photoregressed mammalian seminiferous epithelium, with particular attention to rodents and humans.

Imbalances in the proliferation and apoptosis processes are involved in numerous epithelial alterations. In the seminiferous epithelium, normal spermatogenesis is regulated by spermatogonia proliferation and germ cell apoptosis, and both processes are involved in diverse pathological alterations of the seminiferous epithelium. Other physiological phenomena including aging and short photoperiod, in which apoptosis and proliferation seem to play important roles, cause testicular changes. Aging is accompanied by diminished proliferation and increased apoptosis, the latter occurring in specific states of the seminiferous cycle and considered the cause of epithelium involution. However, there is no clear evidence concerning whether proliferation decreases in the spermatogonia themselves or is due to an alteration in the cell microenvironment that surrounds them. As regards the factors that regulate the process, the data are scant, but it is considered that the diminution of c-kit expression in the spermatagonia, together with the diminution in antiapoptotic factors (Bcl-x(L))) of the intrinsic molecular pathway of apoptosis play a part in epithelial regression. A short photoperiod, especially in rodents, produces a gradual involution of the seminiferous epithelium, which is related with increased apoptosis during the regression phase and a diminution of apoptosis during recrudescence. Proliferative activity varies, especially during the total regression phase, when it usually increases in the undifferentiated spermatogonia. In other species showing seasonal reproduction, however, decreased proliferation is considered the main factor in the regression of the seminiferous epithelium. Little is known about how both phenomena are regulated, although data in rodents suggest that both the intrinsic and extrinsic pathways of apoptosis contribute to the increase in this process. In conclusion, regression of the seminiferous epithelium in physiological situations, as in many pathological situations, is a result of alterations in equilibrium between the proliferation and apoptosis of germinal cell types. However, both physiological phenomena showed important differences as regard proliferation/apoptosis and their regulation pathways, probably as a result of their irreversible or reversible character.

Differences in the response in the dermis of the tails of young and old SD rats to treatment with bipolar RF.

BACKGROUND: The passing of the years is marked by intrinsic (chronological) and extrinsic aging, caused by photoaging, which is characterized by a decrease in collagen and the deposition of abnormal elastic fibers in the dermis. The use of bipolar radiofrequency (RF) increases fibroblast proliferation and differentiation, accompanied by collagen synthesis and a subsequent increase in connective tissue, and it is not known whether the biological effects of this type of radiofrequency on the dermis are similar regardless of the age of the individual or whether such effects are altered by the aging process itself. AIMS: The objective was to perform a histological study of the changes in the tail dermis of young and old rats after submitting them to bipolar RF, to determine cell proliferation and volume of connective tissue. METHODS: One part of the rat tail was fixed in formol and processed for light microscopy and another part processed for electron microscopy. RESULTS: The number of fibroblasts/unit area and cells positive to nuclear proliferation antigen was higher in young animals. Significant differences were observed regarding expression of HSP-47 protein, and the value was always lower in old rats. No significant differences were observed in the percentage of connective tissue. No histological alterations were observed in any rats. CONCLUSION: Treatment with RF increased the number of fibroblasts located in the connective tissue of the young rats. In addition, the effect of a single treatment on the population of fibroblasts in young animals was sufficient to activate the synthesis of new collagen.

[Frequency of the IVS4+66A-G polymorphism in the alpha-synuclein gene in patients with Parkinson's disease in north-western Mexico]. / Frecuencia del polimorfismo IVS4+66A-G en el gen alfa-sinucleína en pacientes con enfermedad de Parkinson del noroccidente de México.

INTRODUCTION: Parkinson disease (PD) is the second most common neurodegenerative disease of adult onset. Is a progressive movement disorder including tremor, bradykinesia, rigidity and postural instability, with an age onset between 43 and 66 years. Histopathologically, is characterized by a severe loss of dopaminergic neurons in the substantia nigra and inclusions consisting of insoluble protein aggregates called Lewy bodies, this are comprised in part of alpha-synuclein. The etiology of PD is still not fully understood, but genetic analyses, epidemiologic studies and experimental models of PD are providing important new insights into the pathogenesis of PD. AIM: To determine allelic and genotypic frequencies of polymorphism IVS4+66A-G in the alpha-synuclein gene and to demonstrate its association with PD in northwest Mexican population. SUBJECTS AND METHODS: Genomic desoxyribonucleic acid (DNA) from 51 PD patients and 121 persons without PD were achieved by polymerase chain reaction and analyzed the allelic and genotypic distribution in IVS4+66A-G polymorphism of alpha-synuclein gene. RESULTS: The genotypic frequency of IVS4+66AA was 43.1% in PD patients and 38.8% in control group; IVS4+66GG was 2% in PD patients and 4.1% in control group, whereas 54.9% in PD patients and 57.1% in control group were heterozygous. Statistical differences were not observed between groups (p<0.05). CONCLUSIONS: Association was not observed between the IVS4+66A-G polymorphism and PD.

Assessment of mechanical behavior of PLA composites reinforced with Mg micro-particles through depth-sensing indentations analysis.

This work deals with the mechanical characterization by depth-sensing indentation (DSI) of PLLA and PLDA composites reinforced with micro-particles of Mg (up to 15wt%), which is a challenging task since the indented volume must provide information of the bulk composite, i.e. contain enough reinforcement particles. The composites were fabricated by combining hot extrusion and compression moulding. Physico-chemical characterization by TGA and DSC indicates that Mg anticipates the thermal degradation of the polymers but does not compromise their stability during processing. Especial emphasis is devoted to determine the effect of strain rate and Mg content on mechanical behavior, thus important information about the visco-elastic behavior and time-dependent response of the composites is obtained. Relevant for the intended application is that Mg addition increases the elastic modulus and hardness of the polymeric matrices and induces a higher resistance to flow. The elastic modulus obtained by DSI experiments shows good agreement with that obtained by uniaxial compression tests. The results indicate that DSI experiments are a reliable method to calculate the modulus of polymeric composites reinforced with micro-particles. Taking into consideration the mechanical properties results, PLA/Mg composite could be used as substitute for biodegradable monolithic polymeric implants already in the market for orthopedics (freeform meshes, mini plates, screws, pins, …), craniomaxillofacial, or spine.

Nanomechanical characterization of alumina coatings grown on FeCrAl alloy by thermal oxidation.

This work studies the feasibility of using repetitive-nano-impact tests with a cube-corner tip and low loads for obtaining quantitative fracture toughness values in thin and brittle coatings. For this purpose, it will be assumed that the impacts are able to produce a cracking, similar to the pattern developed for the classical fracture toughness tests in bulk materials, and therefore, from the crack developed in the repetitive impacts it will be possible to evaluate the suitability of the classical indentation models (Anstins and Laugier) for measuring fracture toughness. However, the length of this crack has to be lower than 10% of the total coating thickness to avoid substrate contributions. For this reason, and in order to ensure a small plastic region localized at the origin of the crack tip, low load values (or small distance between the indenter tip and the surface) have to be used. In order to demonstrate the validity of this technique, repetitive-nano-impact will be done in a fine and dense oxide layer (α-Al2O3), which has been developed on the top of oxide dispersion strengthened (ODS) FeCrAl alloys (PM 2000) by thermal oxidation at elevated temperatures. Moreover, it will be shown how it is possible to know with each new impact the crack geometry evolution from Palmqvist crack to half-penny crack, being able to study the proper evolution of the different values of fracture toughness in terms of both indentation models and as a function of the strain rate, ε̇, decreasing. Thereby, fracture toughness values for α-Al2O3 layer decrease from ~4.40MPam , for high ϵ̇ value (10(3)s(-1)), to ~3.21MPam, for quasi-static ϵ̇ value (10(-3)s(-1)). On the other hand, ϵ̇ a new process to obtain fracture toughness values will be analysed, when the classical indentation models are not met. These values are typically found in the literature for bulk α-Al2O3, demonstrating the use of repetitive-nano-impact tests which not only provide qualitative information about fracture resistance of the materials but it also can be used to obtain quantitative information as fracture toughness values in the case of brittle materials.

Dynamic nanomechanical properties of novel Si-rich intermetallic coatings growth on a medical 316 LVM steel by hot dipping in a hypereutectic Al-25Si alloy.

This aim of this study is to determine the elastoplastic properties of Ni-free Al3FeSi2 intermetallic coatings grown on medical stainless steel under different experimental conditions. Elastoplastic properties are defined by the plasticity index (PI), which correlates the hardness and the Young's modulus. Special emphasis is devoted to correlate the PI with the wear resistance under sliding contact, determined by scratch testing, and fracture toughness, determined by using a novel method based on successive impacts with small loads. With regard to the substrate, the developed coatings are harder and exhibit a lower Young's reduced modulus, irrespective of the experimental conditions. It has been shown that preheating of the samples prior to hot dipping and immersion influences the type and volume fraction of precipitates, which in turn also affect the nanomechanical properties. The higher the preheating temperature is, the greater the Young's reduced modulus is. For a given preheating condition, an increase of the immersion time yields a decrease in hardness. Although apparent friction coefficients of coated specimens are smaller than those obtained on AISI 316 LVM, they increase when using preheating or higher immersion times during processing, which correlates with the PI. The presence of precipitates produces an increase in fracture toughness, with values greater than those presented by samples processed on melted AlSi alloys with lower Si content (12 wt%). Therefore, these intermetallic coatings could be considered "hard but tough", suitable to enhance the wear resistance, especially when using short periods of immersion.

Testicular histomorphometry and the proliferative and apoptotic activities of the seminiferous epithelium in Syrian hamster (Mesocricetus auratus) during regression owing to short photoperiod.

During the non-breeding season some animals exhibit testicular atrophy, decreased testicular weight and reduced seminiferous tubule diameter accompanied by depletion of the seminiferous epithelium. Some cellular factors involved in this depletion are changes in germ cell proliferation and apoptosis. In the Syrian hamster this depletion has been studied histologically and in terms of the involvement of proliferation and apoptosis in the seminiferous epithelium of fully regressed testes. The objectives of this study included the histomorphometrical characterization of the testis and the determination of the proliferative and apoptotic activity of germ cells in the seminiferous epithelium during testicular regression owing to short photoperiod. The study was performed using conventional light microscopy (hematoxylin and eosin), 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 established regression groups: mild regression (MR), strong regression (SR), and total regression (TR). Morphometrically a gradual decrease in total tubular area and in the testicular, tubular, and epithelial volumes was observed during testicular regression. Interstitial and luminal volumes decreased from the MR group onwards. The tubular length decreased from MR to SR. As regards spermatogonial proliferation, only an initial decrease in proliferative activity was observed, whereas apoptotic germ cell activity increased throughout regression. The number of germ cells studied decreased throughout the process of testicular regression. In conclusion, testicular regression in Syrian hamster comprises two histomorphometrical phases, the first involving a decrease in seminiferous tubular diameter and volume and the second involving shortening of the seminiferous tubule and a decrease in interstitial volume. At the cellular level, there is an initial decrease in proliferation and increase in apoptosis involving all germ cells. At the end of regression, the proliferative and apoptotic activities of the spermatogonia recover the values observed prior to regression in preparation for recrudescence.

Cell proliferation in the seminiferous and epididymal epithelia of Sus domesticus.

It is important to understand the proliferative activity of the different structures of the male reproductive apparatus in livestock species, such as Sus domesticus, to ensure reproductive efficiency. The main aims of this study were (a) to evaluate the proliferative activity of the spermatogonia in the different stages of the seminiferous cycle and (b) to study the cell proliferation in the epididymal epithelium in each region, identifying the different cells involved. For this, the testes and epididymis of three healthy, sexually mature Sus domesticus boars were used. The organs were processed for light microscopy, and immunohistochemical techniques were used to detect proliferating cell nuclear antigen. The cells immunostaining positively and negatively for proliferating cell nuclear antigen were counted and several parameters and indexes were calculated to evaluate the proliferation in both epithelia, taking into account the stage of the seminiferous epithelium cycle, and, in the case of the epididymal epithelium, the different regions and cells are the same. Finally, a contrast analysis of equality between pairs of means was carried out followed by a least significant differences test, in which differences were considered significant at P < 0.05. In the seminiferous epithelium, the greatest total number of spermatogonia and proliferating spermatogonia was observed in the postmeiotic stages (mainly VII and VIII). The proliferation index of the spermatogonia increased from the meiotic to postmeiotic stages. As regards the epididymal epithelium, the total proliferation index was higher in the caput. In each region, the clear and principal cells showed the highest proliferation index with respect to the total number of cells counted, whereas the proliferation index of each cell with respect to the same type was higher in the clear cells, followed by the narrow and principal cells. In conclusion, the proliferative activity of spermatogonia in the seminiferous epithelium of Sus domesticus is stage-dependent, and mainly occurs in the postmeiotic stages. In the epididymal epithelium, proliferative activity takes place in several cell types and is dependent on the anatomical region of the epididymis. We think that these results may be of importance for understanding the pathologic or reproductive processes in which cell proliferation is involved in the male reproductive system.

Characterization of the elastic-plastic behavior of intermetallic coatings growth on medical stainless steel by instrumented ultramicroindentation.

The purpose of this work is to study the elastoplastic properties of novel intermetallic coatings grown by hot dipping on medical steel 316 LVM and their correlation with the scratch resistance by means of instrumented ultramicroindentation techniques. Elastoplastic properties are defined by the plasticity index (PI), which correlates the hardness and the Young's modulus, and the yield strength that delimits the elastic-plastic transition. To avoid overestimations of the PI due to the so-called indentation size effect, macroscopic hardness values were determined. The PI of the coating increases from 0.6 to 0.8 with increasing immersion time. These values are always lower than those of the bare substrate (0.9) but higher than those found for ceramics (∼0.5). The increase of the PI is accompanied by a decrease in the yield strength from 0.73 to 0.34 GPa, which highlights the relevance of the compressive residual stresses and their relaxation with increasing immersion time. The higher plasticity is shown by higher apparent friction coefficients (0.159), which are always lower than those of the bare steel (0.264). Therefore, these intermetallic coatings could be considered "hard but tough" coatings, suitable for enhancing the wear resistance of the medical steel, especially when using short periods of immersion. The study provides evidence that the load-depth curve of indentation contains abundant information and that its analysis can be used to determine various mechanical properties of coatings that could be important for load bearing components.

Histological changes in connective tissue of rat tails after bipolar radiofrequency treatment.

Radiofrequency (RF) has been included in the techniques used in aesthetic surgery/medicine. To date, no studies have performed a histological assessment of changes in the tissue after application of bipolar radiofrequency (BRF) with low energy and frequency. The aim of this study was to examine changes that are produced in connective tissue, principally in the fibroblasts, following BRF treatment. Four groups of rats received a different number of RF sessions (1, 2, 3 and 5). The following parameters were determined: the number of fibroblasts/unit area (FA), the proliferation index (PI), the Heat shock Protein 47 index (HSPI) and the percentage of connective tissue (PC). For statistical analysis, two subgroups (A and B) were made for the variables FA, PI and PC, and another two subgroups (C and D) for the variable HSPI. Significant differences for FA, PI and PC were observed between subgroups A and B, FA and PI having higher values in A, while PC had higher values in B. The HSPI in subgroup C showed significantly higher values than in D. Low energy and frequency BRF led to an increase in the number, proliferation and biosynthetic activity of fibroblasts. The resulting stress suffered by fibroblasts as a result of heat may be associated with the phenomenon of hormesis.

Revisión crisis parainfecciosas de enero del 2012 a marzo del 2014 / Review of para-infectious seizures from January 2012 to March 2014

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