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Reproductive Biology and Endocrinology
Attenuation of sleep deprivation dependent deterioration in male fertility parameters by vitamin C
Nermin I. Rizk1,
Mohamed S. Rizk2,
Asmaa S. Mohamed3 &
Yahya M. Naguib ORCID: orcid.org/0000-0001-5851-72381
Reproductive Biology and Endocrinology volume 18, Article number: 2 (2020) Cite this article
Male fertility is multifaceted and its integrity is as well multifactorial. Normal spermatogenesis is dependent on competent testicular function; namely normal anatomy, histology, physiology and hormonal regulation. Lifestyle stressors, including sleep interruption and even deprivation, have been shown to seriously impact male fertility. We studied here both the effects and the possible underlying mechanisms of vitamin C on male fertility in sleep deprived rats.
Thirty male Wistar albino rats were used in the present study. Rats were divided (10/group) into: control (remained in their cages with free access to food and water), sleep deprivation (SD) group (subjected to paradoxical sleep deprivation for 5 consequent days, rats received intra-peritoneal injections of vehicle daily throughout the sleep deprivation), and sleep deprivation vitamin C-treated (SDC) group (subjected to sleep deprivation for 5 consequent days with concomitant intra-peritoneal injections of 100 mg/kg/day vitamin C). Sperm analysis, hormonal assay, and measurement of serum oxidative stress and inflammatory markers were performed. Testicular gene expression of Nrf2 and NF-κβ was assessed. Structural changes were evaluated by testicular histopathology, while PCNA immunostaining was conducted to assess spermatogenesis.
Sleep deprivation had significantly altered sperm motility, viability, morphology and count. Serum levels of cortisol, corticosterone, IL-6, IL-17, MDA were increased, while testosterone and TAC levels were decreased. Testicular gene expression of Nrf2 was decreased, while NF-κβ was increased. Sleep deprivation caused structural changes in the testes, and PCNA immunostaining showed defective spermatogenesis. Administration of vitamin C significantly countered sleep deprivation induced deterioration in male fertility parameters.
Treatment with vitamin C enhanced booth testicular structure and function in sleep deprived rats. Vitamin C could be a potential fertility enhancer against lifestyle stressors.
Infertility is a reasonably common condition with medical, psychological, and financial consequences. Infertility can be defined as the inability of a couple to conceive after 1 year of attempting conception. Infertility affects an estimated 15% of couples worldwide, of which, male are considered to be solely responsible for 20–30% of infertility cases [1]. Accumulating data suggest that there is a progressive reduction in human sperm quality and 50–60% reduction in sperm counts in men in recent decades [2]. Male infertility can be influenced by environmental, occupational, and modifiable lifestyle factors such as psychological stress, obesity, smoking, mobile phone radiations and lack of sleep [3, 4].
Sleep is a physiological periodic state of rest. Sleep is a bio-vital phenomenon that is associated with neuro-endocrin and immunity changes [5]. Adequate sleep is a basic inquiry for healthy life and proper fertility; there is a strong correlation between adequate sleep and gonadotropin releasing hormone (GnRH) secretion which plays a basic role in the productive functions [6]. Furthermore, adequate sleep positively affects the sexual behaviour. It was reported that increasing the night sleep 1 h enhances the sexual activity by 14% [7]. Sleep deprivation (SD) is a common social stress that affects a wide range of population. According to the National Sleep Foundation, there is a marked increase in the incidence of sleep deprivation in the last few years. Night shift workers and patient suffering psycho-social disturbances are the most vulnerable populations. SD involves a wide range of disorders such as; behaviourally induced insufficient sleep syndrome, sleep apnea and insomnia [8].
SD has serious adverse effects on different body functions resulting in cardiovascular diseases, immune disturbances and neuro-endocrinal changes [9]. Furthermore, SD and psychological stress alter the activity of the hypothalamic-pituitary-adrenal (HPA) axis and the sympathetic nervous system with negative impact on both sexuality and fertility [10]. Inadequate sleep has been reported to decrease semen quality [11]. Previously published studies have shown that the immune function could be impaired by sleep deprivation [12]. The level of the immunoglobulins G, A and M was enhanced in a sleep deprived cohort study, suggesting that the serum humoral immunity parameters could be altered following insufficient sleep [13]. It was reported that short sleep duration, long sleep duration and late bedtime impair semen quality partly via the increased production of seminal anti-sperm antibody [14]. One could produce antibodies to his own sperms in certain conditions such as varicocele [15], intercourse [16], as well as testicular inflammation [17]. Spermatogenesis is an active replecative process generating about 1000 sperm/second. The high rate of cell division requires rationally higher mitochondrial oxygen consumption [18]. Under stressful conditions, spermatozoa generate small amounts of reactive oxygen species (ROS). Minimal amounts of ROS are essential for acrosomal reaction and fertilization, however, excessive production of ROS can cause damage of normal spermatozoa through lipid peroxidation and DNA damage [19]. Testicular membrane is rich in polyunsaturated fatty acids (PUFA) rendering the testes vulnerable to lipid peroxidation and eventually oxidative stress injury [18].
Great attention has been given to molecules with potentially polymodal protective effects. Vitamin C, ascorbic acid, is present in the testes presumably playing a pivotal role in the testicular antioxidant defence system and, therefore, supporting spermatogenesis. However, in order to function effectively as an antioxidant, vitamin C must be maintained at high levels in the body [13]. In addition, vitamin C has potential anti-inflammatory properties; vitamin C has been reported to alleviate the inflammatory status by reducing hsCRP and IL-6 in hypertensive and/or diabetic obese patients [20]. On the basis of these considerations, the aim of the present study was to test the hypothesis that vitamin C could counteract the detrimental effects of SD on male fertility. To achieve that, we examined the effect of vitamin C administration on semen quality, reproductive hormones, oxidative and inflammatory markers, testicular structure, and testicular expression of genes contributing to oxidative and inflammatory homeostasis in sleep-deprived adult male rats.
Thirty male Wistar albino rats were used in the present study. The experimental procedures were conducted in adherence to the Guiding Principles in the Use and Care of Animals published by the National Institutes of Health (NIH Publication No 85–23, Revised 1996). Animal care and use was approved by the Menoufia University Ethics Committee. Animals were kept for 10 days prior to the start of the study to allow proper acclimatization. The animals were fed standard laboratory chow and allowed free access to water in an air-conditioned room with a 12 h light-dark cycle.
Following acclimatization, rats were assigned randomly into three experimental groups of 10 rats each:
Control group (C): rats remained in their cages with free access to water and balanced diet.
Sleep deprivation group (SD): rats were subjected to paradoxical sleep deprivation for 5 consequent days. Rats had water and food ad libitum during the sleep deprivation period. Rats received intra-peritoneal injections of vehicle daily throughout the sleep deprivation phase.
Sleep deprivation + vitamin C-treated group (SDC): rats were subjected to sleep deprivation for 5 consequent days with concomitant intra-peritoneal injections of 100 mg/kg/day vitamin C (20% vials, Global Cosmetic Solutions, SL, Spain). Rats had water and food ad libitum during the sleep deprivation period.
Sleep deprivation was induced according to the method of Choi et al., 2016 with slight modifications [9]. Rats were kept in a custom-made glass tank (120 × 40 × 40 cm) containing 10 platforms. The platforms were carefully designed to allow alert standing of each rat, but do not allow them to sleep. When rats tend to fall asleep, they lose their balance; hence they fall in water and awaken. Animals could move only by jumping from one platform to another. Before filling the glass tank with water, rats were left in the glass tank 1 h/day for 3 consequent days for acclimatization. After the acclimatization period, the glass tank was filled with water 3 cm beneath the surface of the platforms.
Blood sample collection
At the end of the study, all rats were fasted overnight. Blood was drawn from each rat via cardiac puncture. The blood was allowed to coagulate for 30 min at room temperature. Blood samples were then centrifuged at 4000 rpm for 15 min to separate serum samples. Serum samples were stored at − 20 °C. Finally, all rats were scarified by cervical dislocation.
Biochemical assessment
Serum levels of cortisol (BioVision, USA), testosterone (CUSABIO, Shanghai, Chaina), interleukin 17 (IL-17, Abcam, USA), and interleukin 6 (IL-6, Abcam, USA) were determined by quantitative sandwich enzyme immunoassay technique using an automatic optical reader (SUNRISE Touchscreen, TECHAN, Salzburg, Austria). Malondialdehyde (MDA) and total antioxidant capacity (TAC) (Abcam, USA) were determined by routine kinetic and fixed rate colorimetric methods on a Jenway Genova autoanalyser (UK).
Evaluation of testicular GSH, MDA and GPx
Rats were sacrificed by cervical dislocation. Both testes were dissected, weighed and then washed with cold saline. The left testes were homogenized in lysis buffer solution (abcam, USA, 1:5 w/v). The homogenate was centrifuged and the supernatant was used for colorimetric estimation of glutathione (GSH, QuantiChrom™, BioAssay Systems, USA), glutathione peroxidase (GSH-Px, EnzyChrom™, BioAssay Systems, USA) and MDA tissue levels using fixed rate colorimetric method.
Collection of semen
Cauda epididymis was dissected free in a Petri dish containing 5 ml warm saline solution (37 °C). Then, it was cut into pieces by a fine medical scissor and incubated for 5 min with frequent shaking to yield semen suspension. Semen suspension was used for further assessment of sperm motility (%), viability (%), abnormal forms (%) and total sperm count (in millions) [9].
Assessment of sperm motility
After 5–10 min of dissection, a drop of semen suspension was smeared on a glass slide and examined under light microscope (power 400X) to assess sperm motility. A minimum of three different fields were examined to determine the mean percentage of sperm motility [21].
Sperm viability
Equal volumes (100 μl) of semen suspension and eosin stain (1%) were mixed and incubated for 2 min. A drop of this mixture was smeared on a clean glass slide and examined under light microscope. Living sperms were not affected by the stain, while dead sperms stained pink by eosin [22]. The percentage of viability was determined in the field examined (number of alive sperm/total number sperm*100). In each sample, a minimum of three different fields were examined to determine the mean percentage of sperm viability.
Assessment of sperm morphology
A drop of semen suspension was smeared on a glass slide and examined by light microscope (power 400X). The percentage of abnormal forms, in each field, was determined (number of abnormal sperms/total number of sperms*100). Ten fields were examined in each slide to determine the mean percentage of abnormal forms [23].
Ten microliter of the semen suspension was smeared on the counting haemocytometer. Sperm counting was done under light microscope (200X). The results were expressed as million/ml of suspension. The sperm count was repeated at least twice and the average was taken. Total sperm count was calculated as (Count*dilution* 5*104) [24].
Histopathology examination
Specimens from the right testes were fixed in 10% formol saline for 5–7 days. The specimens were washed in tap water for 10 min and then dehydrated in graded ethanol solutions (70, 90% over night and 100% ethanol solution for three changes 1 h each). The specimens were cleared in xylene (20–30 times). After that, specimens were impregnated in soft paraffin wax at 55–60 °C for 2 h then in hard paraffin wax at room temperature in moulds. Tissue blocks were cut into section of 5 μm thickness by using rotator microtome. Tissue sections were dipped in a warm water-bath, picked up on clean slides, and placed on hot plate for 2 min. Finally, tissue sections were stained with haematoxylin and eosin stain for general architecture of the studied tissues.
PCNA immunostaining
Immunostaining staining was carried out using primary antiserum to proliferating cell nuclear antigen (PCNA) (PC10, Santa Cruz Biotechnology Inc., Heidelberg, Germany). Briefly, the primary antibody was diluted in Tris buffer with a dilution of 1:50 (as determined by the data sheet). The sections were incubated with the primary antibody overnight at + 4 °C. The binding of the primary antibody was observed using a commercial avidinbiotin- peroxidase detection system recommended by the manufacturer (DAKO, Carpenteria, USA). Finally, the slides were stained with diaminobenzene (DAB).
Analysis of gene expression by quantitative RT-PCR (qRT-PCR)
Real time quantitative reverse transcription-polymerase chain reaction (RT-PCR) assay was used to examine mRNA expression of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and nuclear factor kappa beta (NF-κβ) genes in the studied groups. To extract RNA, frozen testicular specimens were ground using a mortar and pestle and liquid nitrogen. Total RNA was extracted with TRI reagent (Sigma-Aldrich, New South Wales, Australia). To generate the template for PCR amplification, 2 μg of testicular RNA was reverse transcribed into cDNA using the high capacity RNA-to-cDNA kit (Applied Biosystems, Foster City, CA, USA). This cDNA was used to determine the mRNA expression for the genes of interest by quantitative real-time PCR as previously described using gene specific primers (Table 1), designed using Primer Express Software version 2.0 (Applied Biosystems, Victoria, Australia). GAPDH was used as the housekeeping control loading gene. SYBR green PCR assays for each target molecule and internal reference GAPDH were performed in duplicate on these cDNA samples in a 10 μL reaction using Applied Biosystems 7500 FAST 96-well PCR machine. From the amplification curves, relative expression was calculated using the comparative Ct (2 − ΔCt) method, with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) serving as the endogenous control and the expression data as a ratio (target gene/GAPDH).
Table 1 Sequence of the primers used for assessing Nrf2 and NF-κβ gene expression
Kolmogorov-Smirnov test was performed on all data sets to ensure normal distribution (p > 0.5). Results are expressed as mean ± standard deviation (SD). Analyses of Variances (ANOVA) with Tukey's honesty significant difference (HSD) tests were used for statistical analysis using Origin® software and the probability of chance (p values). P values < 0.05 were considered significant.
Although there was no significant difference in body weight between the experimental groups at the start of the experiment; the body weight was significantly lower in the SD group when compared to the control group (142.31 ± 0.89 g vs. 181.66 ± 1.85 g, p < 0.05) following 5 days of sleep deprivation. Interestingly, administration of vitamin C resulted in significant increase in body weight in the in SDC group (153.99 ± 3.02 g, p < 0.05) when compared to the SD group, albeit it was still significantly lower if compared to the corresponding values in the control group, or the SDC group itself at the start of the experiment (Fig. 1a). Unsurprisingly, there was significant increase in the testicular index in the SD group when compared to the control group (0.54 ± 0.018 vs. 0.44 ± 0.012, p < 0.05), while there was significant reduction in the SDC group (0.492 ± 0.016, p < 0.05) when compared to SD group. Testicular index was significantly higher in the SDC group when compared to the control group (Fig. 1b).
Effect of vitamin C on body weight, testicular index and sperm evaluation in sleep deprived rats. a Body weight in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. b Testicular index in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. c Sperm count in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. d Sperm viability in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. e Sperm motility in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. f Abnormal forms in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. (Significant = p < 0.05, * significant when compared to the control group, • significant when compared to the sleep deprived group, # significant when compared to the same group at the start of the experiment. Number of rats = 10/group)
As shown in Fig. 1c, d, e, and f, there was significant decrease in sperm count, viability and motility, with significant increase in abnormal forms of sperms in SD group when compared to the control group (16.15 ± 1.07 × 106, 70.6 ± 1.96%, 80 ± 2.74% and 15.6 ± 1.33% vs. 49.42 ± 2.88 × 106, 93.2 ± 0.97%, 94 ± 2.45% and 7 ± 0.95% respectively, p < 0.05). Sperm count, viability and motility were significantly higher, while the abnormal forms of sperms were significantly lower in the SDC group (38.86 ± 4.34 × 106, 87.6 ± 1.12%, 88 ± 2.55% and 11.4 ± 0.68% respectively, p < 0.05) compared to the SD group. However, sperm count, viability and motility still significantly lower and abnormal forms still significantly higher in SDC group when compared to control group.
Serum cortisol and corticosterone levels were significantly higher, while testosterone level was significantly lower in the SD group when compared to the control group (212.58 ± 18.44 ng/ml, 224.6 ± 8.12 ng/ml and 1.79 ± 0.14 ng/ml vs. 72.15 ± 6.98 ng/ml, 52.4 ± 3.17 ng/ml and 3.95 ± 0.11 ng/ml respectively, p < 0.05). In the vitamin C treated sleep-deprived rats, serum cortisol and corticosterone levels were significantly lower, while testosterone level was higher (90.43 ± 9.35 ng/ml, 73.48 ± 9.36 ng/ml and 3.48 ± 0.25 ng/ml respectively, p < 0.05) when compared to the SD group. However, cortisol and corticosterone levels were significantly higher and testosterone level was significantly lower in the SDC group when compared to the control group (Fig. 2).
Effect of vitamin C on serum hormones in sleep deprived rats. a Cortisol level in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. b Corticosterone level in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. c Testosterone level in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. (Significant = p < 0.05, * significant when compared to the control group, • significant when compared to the sleep deprived group. Number of rats = 10/group)
Serum levels of IL-6 and IL-17 were significantly higher in the SD group when compared to the control group (26.6 ± 1.6 pg/ml and 46.1 ± 3.16 pg/ml vs. 13.4 ± 0.51 pg/ml and 23 ± 1.41 pg/ml respectively, p < 0.05). IL-6 and IL-17 levels were significantly lower in the SDC group (19.8 ± 0.58 and 31.4 ± 1.33 pg/ml) when compared SD groups, yet they were still significantly higher than the corresponding values in the control group (Fig. 3a and b). Sleep deprivation resulted in significant reduction in the TAC and significant elevation in MDA levels when compared to the control group (0.79 ± 0.01 mM/ml and 2.05 ± 0.09 uM/ml vs. 1.05 ± 0.05 mM/ml and 0.46 ± 0.07 uM/ml respectively, p < 0.05). TAC was significantly higher and MDA was significantly lower in the SDC group (0. 9 ± 0.05 mM/ml and 1.04 ± 0.06 uM/ml respectively, p < 0.05) when compared to the SD group, while the TAC remained significantly lower with significantly higher MDA levels in the SDC group if compared to the control group (Fig. 3c and d).
Effect of vitamin C on serum inflammatory and oxidative stress markers in sleep deprived rats. a IL-6 level in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. b IL-17 level in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. c TAC in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. d MDA level in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. (Significant = p < 0.05, * significant when compared to the control group, • significant when compared to the sleep deprived group. Number of rats = 10/group)
Effect of vitamin C on testicular oxidative-antioxidative parameters in sleep deprived rats. a GSH level in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. b MDA level in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. c GPx level in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. (Significant = p < 0.05, * significant when compared to the control group, • significant when compared to the sleep deprived group. Number of rats = 10/group)
Testicular GSH and GPx tissue levels were significantly lower, while testicular MDA was significantly higher in the SD group when compared to the control group (18.15 ± 3.63 uM/g tissue, 83.47 ± 12.33 U/g tissue and 74.37 ± 2.93 ng/g tissue vs. 46.67 ± 4.41 uM/g tissue, 134.12 ± 18.79 U/g tissue and 34.81 ± 0.94 ng/g tissue respectively, p < 0.05). In vitamin C treated SD group, testicular tissue levels of GSH and GPx were significantly higher, while MDA tissue level was significantly lower (33.87 ± 3.92 uM/g tissue, 152.92 ± 21.72 U/g tissue and 42.02 ± 1.77 ng/g tissue respectively, p < 0.05) when compared to the SD groups. Testicular level of GSH was significantly lower, while MDA level was significantly higher in the SDC group, when compared to the control group (Fig. 4).
Gene expression of Nrf2 in the testicular tissue was significantly downregulated, while gene expression of NF-κβ gene was significantly upregulated in the SD group when compared to the control group (0.62 ± 0.014 and 1.87 ± 0.02, vs. 1 RQ respectively, p < 0.05). Nrf2 gene expression was significantly higher, while NF-κβ gene expression was significantly lower in the SDC group (0.89 ± 0.06 and 1.29 ± 0.04 RQ respectively, p < 0.05) when compared to the SD group, however, the gene expression of Nrf2 remained significantly lower and NF-κβ significantly higher in the SDC if compared to the corresponding values in the control group (Fig. 5).
Effect of vitamin C on testicular Nrf2 and NF-κβ gene expression in sleep deprived rats. a Nrf2 gene expression in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. b NF-κβ gene expression in control (white column), sleep deprived (black column) and sleep deprived + vitamin C treated (grey column) groups. (Significant = p < 0.05, * significant when compared to the control group, • significant when compared to the sleep deprived group. Number of rats = 10/group)
Histopathological evaluation of testicular biopsies revealed abnormal morphology of seminiferous tubules in the SD group with cellular degeneration of spermatogonia and thickening of the basement membrane. Interestingly, in the SDC group, more preserved architecture and morphology of spermatogonia were observed (Fig. 6). PCNA immunostaining revealed decreased positive immunostaining of the basal cell layer in the SD group when compared to the control group. Treatment with vitamin C resulted in enhancement of PCNA immunostaining when compared to the SD group, reflecting a qualitative improvement of spermatogenesis (Fig. 7).
Vitamin C preserves testicular structure in sleep deprived rats. a Representative photomicrograph of Hx & E stained testis sections from control group showing normal oriented regular shaped seminiferous tubules with different stages of spermatogenesis, thin basement membrane (arrow) and interstitial spaces showing thin walled blood vessels (X 200). b Representative photomicrograph of Hx & E stained testis sections from sleep deprived group showing multiple seminiferous tubules with abnormal morphology of spermatogonia (arrows), shrunken nucleus and vacuolated cytoplasm), occasional apoptotic cells and very few late spermatids. Thick walled blood vessels can be seen in interstitial spaces (X 200). c Representative photomicrograph of Hx & E stained testis sections from sleep deprived + vitamin C treated group showing seminiferous tubules lined by multiple layers of spermatogonia and spermatocytes, with early and late spermatids filling the lumen(X 200). (Number of rats = 10/group)
PCNA immunostaining in the studied groups. a Representative photomicrograph of PCNA immunostained sections in testis of control group showing strong positive staining of most of proliferating basal cells in seminiferous tubules. b Representative photomicrograph of PCNA immunostained sections in testis of sleep deprived group showing patchy positive staining of proliferating basal cells in seminiferous tubules c Representative photomicrograph of PCNA immunostained sections in testis sleep deprived + vitamin C treated group showing increased positive staining of proliferating basal cells in seminiferous tubules. (X 200, Number of rats = 10/group)
Infertility is defined as inability of couples to conceive after 1 year of unprotected intercourse. Thereby, infertility affects 13–18% of couples, and male factor accounts for up to half of all the cases. Development of male infertility is influenced by many diseases and/or risk factors. Importantly, increase in risk of infertility can be noted, mostly in male population, when exposed to environmental stressors including sleep deprivation [25]. Better understanding of fertility and semen quality at the molecular levels in the male reproductive system could lead to a great achievement in treating infertility. Essentially, a better treatment for fertility and sexual dysfunction could improve the overall quality of life. Molecules with polymodal actions have gained much attention to minimize male reproductive tissue injuries, and enhance male fertility.
In the present study, sleep deprivation resulted in significant decrease in sperm count, viability and motility, while there was significant increase in abnormal forms and testicular index. Sleep deprivation was linked to alteration of the quality of sperm [9, 14], however the precise mechanism has not been elucidated. It could be possible that the associated inflammatory and oxidative stress mediators play an injurious role with consequent reduction in sperm quality [26, 27]. Vitamin C has been shown to attenuate male reproductive dysfunction in diabetic rats [28].Vitamin C has also been reported to improve sperm count, motility, progression, viability and anomalies in rats subjected to forced swimming stress [22]. These effects were mainly attributed to the antioxidant and the testosterone increase properties of vitamin C. In our hands vitamin C countered SD-induced injurious effects on sperm characteristics, testis weight and testicular index. We then went to validate the underlying mechanisms, studying the possibility of anti-oxidant, anti-inflammatory, hormone and gene modifying effects.
SD induces intense alterations in the regulatory endocrinal axes, including the hypothalamic-pituitary-adrenal (HPA) axis. In the present study SD resulted in significant increase in serum cortisol and corticosterone levels, while it caused significant decrease in serum testosterone level. Vitamin C opposed the SD-induced hormonal alterations. Despite the stress modality, stress-induced increase in corticosterone and decrease in testosterone levels have been reported [9, 29]. In fact, the decrease in testosterone concentration was attributed to the increase in corticosterone level, as part of the stress-induced activation of the HPA axis, resulting in inhibition of the hypothalamic-pituitary-gonadal (HPG) axis [30]. Elevated corticosterone levels not only decreases testosterone production by Leydig cells, it also induces Leydig cells apoptosis [31, 32]. It has been reported that a negative relationship exists between cortisol and testosterone. Elevated cortisol levels were associated with decreased testosterone levels during exercise or even in disease status such as ischemic heart disease [33, 34]. Vitamin C supplementation has also been reported to attenuate cortisol responses following psychological or physical stressors [35]. Vitamin C is secreted from the adrenals in response to adrenocorticotrophic hormone (ACTH), representing a hormone-regulated paracrine secretion of vitamin C as part of the stress response [36]. Interestingly, and in support to our findings, supplementation with vitamin C attenuated the increase in the blood cortisol, adrenaline, interleukin-10 (IL-10) and interleukin-1 receptor antagonist (IL-1Ra) levels following ultra-marathon running [37]. Furthermore, vitamin C was shown to reduce corticosterone level in non-adrenalectomized rats alleviating stress-related behavior [38].Hence, we could imply that in our study vitamin C boosted testosterone concentration and thereby, improved SD-induced diminution in sperm quality.
It is well documented that oxidative stress is implicated in male factor infertility. In the present study, sleep deprived male rats showed higher serum and testicular tissue levels of MDA, while they had lower serum TAC and testicular tissue GSH and GPx levels when compared to the control group. Administration of vitamin C significantly attenuated sleep deprivation induced alteration in oxidative stress markers. A bidirectional relationship between sleep deprivation and oxidative stress has been documented [39,40,41]. Previous data showed evidence that the pathophysiology of male infertility was highly influenced by impairment in seminal antioxidant and lipid peroxidation status. Lifestyle stress, reduces male fertility; an increasing number of cases of male infertility are thought to be primarily due to oxidative stress [42]. MDA serves as an index of lipid peroxidation and a marker of oxidative stress, and could serve as a diagnostic tool for of infertility in asthenozoospermic patients [43, 44]. MDA level in seminal plasma has been reported to be negatively correlated with sperm viability, motility, morphology and concentration [44]. On the contrary, TAC levels were positively associated with sperm concentration, motility, and morphology [45]. GPx can be considered as a predictive measure for fertilization capacity. In fact, GPx is thought to be essential for structural integrity of spermatozoa, and a significant determinant of sperm motility and viability. Alteration in the content of GPx, irrespective of the cause, is negatively correlated with fertility-related parameter [46]. Glutathione (GSH) synthesis is induced in cells exposed to oxidative stress as an adaptive process. The relationship of GSH enzymatic system with oxidative stress in the ejaculate has gained much attention and the regulation of its activity in the semen has been suggested as a therapeutic strategy. Interestingly, intracellular sperm GSH system is altered in infertile men, which seems to be linked to sperm morphology. The quest to find novel antioxidants and/or combinations developed for safe and efficient treatment of oxidative stress induced infertility is likely to continue. Almost three decades ago, the antioxidant efficacy of vitamin C was shown to be effective for the treatment of sperm oxidative stress in smokers [47]. Since then, only few studies were conducted to confirm this finding. Indeed, vitamin C seems indispensible protector of semen from ROS; semen samples with excess ROS where found to be correlated with very low vitamin C concentrations [48]. Our results support the idea that vitamin C could be an efficient therapeutic option for the treatment of oxidative stress caused by sleep deprivation, and apparently, other environmental stressors via its potent antioxidant properties.
Since sleep deprivation can cause a state of inflammation [49], it was relevant to study the possible effects of inflammation on the male reproductive system. Inflammation has been known to affect the twin testicular functions; steroidogenesis and spermatogenesis. Marked decreases in the circulating levels of luteinizing hormone and testosterone were detected during inflammatory states [50]. Indeed, testicular inflammatory disorders leading to impairment of spermatogenesis are considered to be a primary cause for male infertility. The testis is considered as an immune privileged organ, nevertheless, toxic agents and inflammation may overwhelm immune suppressor mechanisms resulting in autoimmune reactions against spermatic antigens. Consequently, this may result in aspermatogenesis and infertility [51]. In the present study, sleep deprivation resulted in significant rise in IL-6 and Il-17, which could be countered by the treatment with vitamin C. Nevertheless, some cytokines such as IL-1, and IL-6 can also produced by Leydig and Sertoli cells [25]. Consequently, it is possible that cytokines may act not only towards somatic cells, but also towards the germ cells both in an autocrine and paracrine fashion. It is possible that cytokines may act during spermatogenesis, sperm maturation, sperm transport, and even during the fertilization process itself. Cytokines such as interleukins and tumor necrosis factors are involved in signal transduction during inflammatory states [26]. Despite the existing controversy regarding the role of cytokines in fertility, our results were in agreement with previously published data reporting that significantly elevated IL-6 levels were found in infertile patients and revealed an apparent negative correlation with sperm number. Moreover, infertile patients with varicocele exhibited an elevated levels of IL-6 [52, 53]. Excess IL-17 is commonly associated with different types of inflammation, and as for our study, IL-17 serum levels were elevated in the sleep deprived male rats. It has been reported previously that LI-17 and its signaling pathway were highly expressed in mice testis exposed to high fluoride [51]. IL-17 was found to be critically involved in male patients with azoospermic testis with chronic inflammation. To our knowledge, this could be the first report on the effect of Vitamin C on serum IL-6 or IL-17 vis-à-vis male fertility.
In the present study, vitamin C had significantly countered the sleep deprivation-induced increased expression of testicular NF-κβ and decreased expression of testicular Nrf2 genes. NF-κβ can be activated by a multiplicity of stimulants including ROS through the phosphorylation of the inhibitory kappa B (IκB) by IκB kinases. NF-κβ is known to activate several genes including the inducible nitric oxide synthase (iNOS), resulting eventually in excessive generation of nitric oxide (NO) [54]. NO, if oxidized, generates reactive NO species, which might behave similarly to ROS. It has been reported previously that NO could enhance cellular injury by decreasing intracellular GSH levels [55]. Nrf2, a redox sensitive transcription factor, is a fundamental contributor to oxidative stress homeostasis [56]. Nrf2 is involved in the regulation of the synthesis and conjugation of glutathione (glutamate-cysteine ligase catalytic subunit), and antioxidant proteins responsible for the detoxification of ROS [57]. It was reported previously that Nrf2 expression is significantly lower in semen of men with low sperm motility [58]. Nrf2 plays an important role in preventing oxidative disruption of spermatogenesis. In fact, Nakamura et al. demonstrated that male Nrf2 knockout mice (Nrf2−/−) have decreased fertility compared to the wild type. They also reported that Nrf2−/− male mice had elevated levels of testicular and epididymal lipid peroxidation, prominent testicular germ cell apoptosis, and reduced antioxidants levels compared to wild type male mice [59].
Histopathological and immunostaining studies demonstrated that vitamin C has protective effects at the structural level. Sleep deprivation resulted in disruption of the normal morphology of spermatogonia and occasionally apoptosis. Treatment with vitamin C retained much of the normal morphology and regularity of the seminiferous tubules and the different stages of spermatogenesis. Preservation of spermatogenesis was further supported by enhancement of the PCNA immunostaining in the sleep deprived vitamin C treated rats. PCNA could serve as a biomarker for spermatogenesis [60].
Sleep deprivation, whatever the cause is, has serious effects on male fertility. We showed here that vitamin C maintained testicular structure and enhanced testicular function in sleep deprived rats. Vitamin C counteracted sleep deprivation dependent alteration in sperm analysis, hormonal levels, and inflammatory and oxidative stress biomarkers. Vitamin C modified the sleep deprivation-dependent altered Nrf2 and NF-κβ gene expression. Consequently, vitamin C could be a potential fertility enhancer in opposition to lifestyle stresses.
Data supporting findings are presented within the manuscript.
ACTH:
Adrenocorticotrophic hormone
GAPDH:
GnRH:
Gonadotropin releasing hormone
GSH:
GSH-Px:
HPA:
Hypothalamic-pituitary-adrenal axis
IL-17:
Interleukin 17
IL-6:
Interleukin 6
MDA:
Malondialdehyde
NF-κβ:
Nuclear factor kappa beta
Nrf2:
Nuclear factor (erythroid-derived 2)-like 2
PCNA:
Polyunsaturated fatty acids
SD:
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Authors wish to thank the Faculty of Medicine - Menoufia University for providing most of the required facilities. Authors would also like to acknowledge the assistance of Prof. Dr. Eman Badr and members of the Central Lab – Faculty of Medicine – Menoufia University.
This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.
Clinical Physiology Department, Faculty of Medicine, Menoufia University, Menoufia, Egypt
Nermin I. Rizk
& Yahya M. Naguib
Medical Biochemistry and Molecular Biology Department, Faculty of Medicine, Menoufia University, Menoufia, Egypt
Mohamed S. Rizk
Pathology Department, Faculty of Medicine, Menoufia University, Menoufia, Egypt
Asmaa S. Mohamed
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NIR performed the sleep deprivation experiments, collected all the samples and participated in drafting the manuscript. MSR performed the biochemical assays and semen analysis. ASM carried out the H&E and immunostaining experiments and interpreted their data. YMN designed the study, prepared the figures, interpreted the data and drafted the manuscript. All authors have read and approved the final draft of the manuscript.
Correspondence to Yahya M. Naguib.
This study was approved by the Ethical Committee of the Faculty of Medicine, Menoufia University, Egypt.
Rizk, N.I., Rizk, M.S., Mohamed, A.S. et al. Attenuation of sleep deprivation dependent deterioration in male fertility parameters by vitamin C. Reprod Biol Endocrinol 18, 2 (2020) doi:10.1186/s12958-020-0563-y
DOI: https://doi.org/10.1186/s12958-020-0563-y
Nrf2


lassical architecture is a universal architecture of precision, planning, and control. Each element has its proper place and size, and each is subordinated to the greater plan. In antiquity, classicism was the architectural language of empire; in the nineteenth century is was the language of manifest destiny and of a Republic taming the wilderness; in the twentieth century, it became the language of fascism.
Bernstein, Mark. Getting Started with Hypertext Narrative. Watertown: Eastgate Systems, 2015.


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http://www.heraldandnews.com/articles/2006/03/09/news/local_news/local2.txt
Dam removal could help Tribes
March 9, 2006 by STEVE KADEL H&N Staff Writer
A key piece of Klamath Tribes' history could live again if removing the Chiloquin dam helps sucker fish thrive.
"These fish have had a subsistence and cultural importance to the Tribes," said Doug Tedrick of the Bureau of Indian Affairs. "They are a natural resource that is important to the Tribes. They have a treaty right to harvest these fish that they are not able to enjoy because they are endangered."
A big step toward removing the dam, built in 1914, comes this summer when a pumping plant will be built next to the Sprague River. It will supply water to irrigators after the dam is gone.
However, removal won't happen for at least another year because the pumping plant must undergo a 60-day test in April 2007.
Tedrick, chief range conservationist for the BIA, was in Klamath Falls this week to talk with Modoc Point Irrigation District representatives and other collaborators about the dam's removal.
But that is just part of efforts to bolster the sucker population. Upstream habitat restoration also is needed to get suckers off the endangered list, Tedrick said.
Dave Northrup of Chiloquin worked on the crew that replaced a fish ladder on the north side of the dam in the 1960s. He visited the dam this month and watched a crew from the U.S. Geologic Survey working in the ladder - tagging and counting fish.
"They told me the suckers are not running yet, but they expect them in a week or two in the ladder," Northrup said. "They did see three large trout in the ladder.
"They also told me they have tracked suckers they have tagged in the ladder as far upstream as the Sycan Marsh."
Northrup would hate to see the dam removed because it creates a popular fishing hole.
Tedrick acknowledges that some fish get past the dam, but not enough to improve sucker numbers.
"Some fish can zip right up, but others have more trouble," he said. "It doesn't block passage, but it impairs passage. It's been identified as one of the causes of their reduced population."
The National Academy of Sciences' research council has given removal of Chiloquin dam high priority as a recovery effort.
Tedrick said environmentalists, the Tribes, and federal, state and local officials all support removal. The BIA and Modoc Point Irrigation District are working together on the project.
"We're not pushing ahead of the district," Tedrick said. "It's their private dam. We've worked hand in hand with the district to make sure they remain whole."
That includes the BIA paying to build the pumping plant as well as for its maintenance. The agency also will pay electricity costs to pump water to Modoc Point, where 5,000 acres are cultivated.
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text:
Bath Village Road Rehabilitation Project advancing; Minister Brand satisfied
February 14, 2019 in NIA
The new asphalted entrance to Bath Village opened to vehicular traffic on February 13, 2019. It is part of the first phase of the Bath Village Road Rehabilitation Project being undertaken by the Nevis Island Administration through the Public Works Department
NIA CHARLESTOWN NEVIS (February 14, 2019) – The Public Works Department (PWD) in the Nevis Island Administration (NIA) completed the first half of Phase 1 of the Bath Village Road Rehabilitation project on February 12, 2019, and work has already begun on the second half which is earmarked for completion in six weeks.
Hon. Spencer Brand, Minister of Public works and Physical Planning in the Nevis Island Administration (file photo)
Hon. Spencer Brand, Minister of Public Works and Physical Planning on Nevis, told the Department of Information on February 13, 2018 that he is pleased with what has transpired so far on the project.
"I am indeed delighted to see that the initial phase of the Bath Village Road Rehabilitation Project was completed. I think that the quality of work we have seen there is certainly what I expected, and certainly surpasses even what many persons would have expected. I am satisfied with the quality.
"I believe that it was a piece of road that in all honesty took a bit longer than we anticipated. We would have seen the relocation of a number of the utilities," he said.
Mr. Brand explained that the Nevis Electricity Company Limited (NEVLEC) relocated poles and electrical wires. The telephone company FLOW, relocated its fibre optic cables and the Nevis Water Department relocated their water main in the area.
He added that apart from the roadway, a number of driveways to various properties were constructed and they were done to the satisfaction of residents there.
Employees from the Public Works Department asphalting the main entrance to Bath Village from Charlestown on February 13, 2019
A replacement u-drain was also constructed to accommodate the anticipated volume of water flowing in that area to the Bath Stream.
Preparatory work on the second section of Phase 1 commenced right after the first section was completed and immediately opened to vehicular traffic on February 13, 2019.
Mr. Brand believes the second section would be completed in quick time.
"We would be continuing going west to Black Hat area, and we anticipate that that portion of the work would be done in a much shorter period than what we had in this initial phase because we don't anticipate the level of construction, as it relates to u-drains and the removal of other utilities and water mains and the like.
"We would get to that area immediately and we will continue to make sure that when all is said and done Bath Village would be looking brand new with their much improved interconnection of roads in the area," he said.
Employees from the Public Works Department working on the second section of Phase 1 of the Bath Village Road Rehabilitation Project on February 13, 2019
Phase 1 is part of the larger project which entails a total of 1,650 feet road which is expected to cost $450,000. The Nevis Island Administration has taken a phased approach for the project's completion.
At a town hall meeting for Bath Village residents in October 2018, to discuss details of the project, Mr. Brand spoke of the sections of road the project is expected to encompass and additional planned road work for the area designed to complement the project.
"My hope is that when this project is completed we will see the completion of rehabilitation work from Reliable Motors junction at the Island Main Road into Bath Village to the roundabout down to Black Hat, Ms. Butler, up the hill by Maude Smith Pre-School all the way over to VON Radio up to Spotlight connecting right next to the good fish lady and the other side of it coming from the roundabout, up the hill again to the dam all the way into Bath Village proper.
Mr. Daniel Williams, Supervisor of Roads at the Public Works Department on Nevis directing road works at Bath Village on February 13, 2019
"We are also envisioning that we will see the completion of the section from the tamarind tree all the way to down to Flexy and the other section from the dam back down to Maude Smith Pre-School. That division that we hope to accomplish once this project would have been completed," he said.
Meantime, Mr. Daniel Williams, Supervisor of Roads at the PWD told the Department of Information that they utilised 225 tons of asphalt for the first section of the project.
He added that vehicular traffic has been flowing through the entrance of Bath Village immediately after the asphalting was completed.
Mr. Williams noted that the use of asphalt mix has a distinct advantage giving way to vehicular traffic with little or no disruption to the motoring public.
and work has already begun on the second half which is earmarked for completion in six weeks.
Bath Village Road Rehabilitation Project
Hon. Spencer Brand
Minister of Public Works and Physical Planning
Nevis Electricity Company Limited
Nevis' Junior Health Brandy-Williams delivers address in observance of World Cancer Day and International Childhood Cancer Day
Team Unity Administration to hold public consultation on Nevis


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LGBTQ BOOKS BY LGBTQ AUTHORS (#OWNVOICES) | Hollieblog
With the rise of the hashtag #ownvoices, we finally have a platform in which we can showcase, promote and support books written about minorities for minorities. It means books with POC characters written by POC authors have been championed and placed solidly on numerous bestseller lists. The same can be said for so many other minorities; I've had the privilege of reading stories on mental illness written by authors who have said mental illness, stories with Muslim characters written by Muslims, and so on. Including LGBTQ+ characters in a story is not a new concept. Whether their sexuality is suggestive or explicit, a centred story line or just a fact about them, it's not uncommon to come across a book and be pleasantly surprised that the world you're reading about doesn't only have straight people in. To celebrate LGBTQ+ History Month, I wanted to list and recommend some YA books that, while include a LGBTQ+ story line or characters, are also #ownvoices. Written by LGBTQ+ folk for LGBTQ+ folk (and everyone else) to enjoy! Disclaimer: I have only included authors who are public and open about their sexuality/gender identity. Sexuality is a private matter for many people who identify as anything but straight, and I don't want this post to create rumour, speculation, or straight up be an invasion of privacy. If you are one of the authors mentioned in this post and do not wish to be included, feel free to email me at hollieblog@outlook.com or through any of my social medias and I'll take your name down! Adam Silvera I could write many love letters to Adam Silvera's books. All of them are centred on an LGBTQ+ character of Latin American descent, and they're always so damn sad but damn beautiful at the same time. They're all standalones, all with their own brand of love and sadness, but here I've ranked them in the order I cried: History is All You Left Me (My tears created a river) They Both Die at the End (My tears removed all my makeup) More Happy Than Not (My tears blurred my vision) V.E. Schwab Ahhhh, there she is. My fave. The one who knows how to simultaneously ruin my life and also make it better. V.E. Schwab has long been advocate for LGBTQ+ books and is not a stranger to including queer characters into her own work. Of course, queer authors are not obligated to write queer characters, and so I've only included the Shades of Magic trilogy which includes A Darker Shade of Magic, A Gathering of Shadows, and A Conjuring of Light. Robin Talley I've recently got into reading Robin Talley's older novels and I love the authentic voice each book has. She's fantastic at telling stories about queer girls whether it's in a haunted boarding school or during segregation in the fifties. I can't wait to read more of her novels, but the ones I have read are Lies We Tell Ourselves and As I Descended. Patrick Ness I think Patrick Ness is considered one of the most prolific YA writers of our generation. His novel, A Monster Calls, was made into a worldwide hit movie, and his sci-fi dystopian trilogy, Chaos Walking, is currently being filmed and will be starring Daisy Ridley and Tom Holland as main characters. But we're here to celebrate Ness' beautiful and powerful novels with some fantastic LGBTQ+ characters! Books I've read include Release, The Rest Of Us Just Live Here, and More Than This. David Levithan David Levithan is a staple in the LGBT YA community with so many books on LGBTQ+ issues, queer characters and story lines, I'm thinking he must be a really fast typer! I haven't exactly read a lot of David Levithan's books, although I feel like I have because he's also collaborated with so many authors that at this point he's literally everywhere. Chances are, you've read a book by David Levithan. Books include Two Boys Kissing, Boy Meets Boy, and Everyday. Nina LaCour I've only read a couple of Nina LaCour's short stories, but every single one I've loved. She writes compelling stories about young queer girls navigating their lives through loss, heartbreak, family, and acceptance. Some of her books include Everything Leads To You and We Are Okay. There are also so many LGBTQ+ #ownvoices novels out there for you to enjoy, support and promote with authors just starting out or have been playing the publishing game for years. Whether or not they've only released the one book or ten, they all deserve a recommendation, and so I've created a small collage of some that have been recommended to me as well as some that I've seen roaming around Book Twitter and Instagram. Dahlia Adler is also doing fantastic work with LGBTQ Reads, a review blog and directory for all your LGBTQ+ rep needs! Of course, this is not the be all end all of LGBTQ+ own voices. If there are any books you would like to share that you think would fit the bill, let me know in the comments! LIKE THIS POST? BUY ME A KO-FI ♥ AMAZON – BOOK DEPOSITORY – WORDERY GOODREADS | INSTAGRAM | TWITTER | TUMBLR | BLOGLOVIN' Other Posts You Might Enjoy


August may be over but this adorable dress for girls pattern is totally worth making this fall too! The cross over bodice is extra cute when made from different fabrics and with a mix of buttons. Before stressing about sewing buttonholes, the buttons are nonfunctional so nothing to worry about.
The free pattern comes in little girl sizes 2-9.


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Home/ Industries/ Food/ Bioscience/ Molecular Biology/ ICGENE Diagnosis and Species Identification / Phytopathology
Phytopathology
The United Nations General Assembly proclaimed 2020 the "International Year of Plant Health" (IYPH). In this way the importance of plants and their health for food security, raw materials, biodiversity and the various contributions by the planet's agricultural, forestry, aquatic and other ecosystems has been recognized.
The control of the health of plants is defined as the discipline in which measures are set and implemented to combat harmful organisms in order to prevent their spread to new areas. In recent years, international trade in plant and seed material has intensified, increasing the risk of introducing pathogenic organisms into new geographical areas. Already today, worldwide losses of yield caused by pests and plant diseases are estimated on average as 21.5% for wheat, 30.0% for rice, 22.6% for maize, 17.2% for potatoes and 21.4% for soybeans. And let's remember that these few crops currently meet half the global calorie requirement. It is clear, then, that common strategies for plant protection measures need to be developed to identify, assess and mitigate these risks.
In Europe, for example, from 14 December 2019, the 'plant passport' which provides for a 'new European plant protection regime' has become mandatory to ensure a homogeneous application of plant health standards in different Member States. To protect the different geographical areas, prevention methods are implemented to ensure the free exchange of plants. The controls, to date carried out by the competent bodies, are made using well-established analytical methods, but these methods are not always adequate and particularly so in the case of emergencies.
In recent years, new technologies and increasingly efficient tests have become available and which meet the new control requirements set by regulatory bodies with the result that such that, in some cases, the use of the ELISA technique has been supplanted by biomolecular techniques.
The kits proposed in this section have been developed by our researchers and are intended for the control of quarantine pathogens, and pathogens that can have important repercussions on the quality and quantity of important crops and products e.g. drupacea, pomacee and wine. All our tests, which are characterized by ease of use, speed and reliability, assist scientists, manufacturers and other operators in carrying out screening tests.
In Europe, for example, as of 14 December 2019, a 'plant passport' has become compulsory, providing for a 'new European plant health regime to ensure the more homogeneous application of plant health rules in the different Member States'. To protect the different geographical areas, prevention is implemented to continue the free trade of plants.
Biosciences General Catalogue
Diagnosis & Species Identification
Price 0,00€


Overall aim and key objectives Advances in optical imaging techniques over the past decades have revolutionized our ability to study chemically reactive flows encountered in air-breathing combustion systems. Emerging technology for unravelling clean- and efficient heat release is needed for advancing new reduced emission technology, and is on the central agenda for a wide variety of energy production- and transport industry. Combustion of fossil fuels remains our largest source of energy production in the world, and global concerns regarding energy security, environmental pollution, and anthropogenic climate change have motivated a large body of research devoted to the experimental measurement and numerical simulation of combustion systems. Clean combustion engineering is the search for improved efficiency by means of strengthen the systems fuel-economy and lowering the emission of NOx, particulates, CO and unburned hydrocarbons (incomplete combustion). New reduced emission technology, greatly rely upon the ability to control the heat release and the exhaust produced by the exothermic reactions between the fuel and the oxidizer in the chemically reactive flow. For the engineering system design, it exist a significant need to inform on the flame-physics involved based on direct observation of the combustion reaction progress and interaction, which is a demanding task for any measurement technique. Chemically reactive flows are inherently multiscale, fully characterized in three-dimensional space and evolving on rapid time-scales. The combustion environment imposes a significant challenge for diagnostics, where it needs to be collected complete information ideally with correlated-field multi-parameter measurement capabilities, exhibiting high spatial and temporal resolution and provided within a snap-shot to freeze the fast dynamics involved. Concurrent detection of major- and minor molecular species (multiplexing) and determining the three most important scalars; the temperature, the flow-field, and the mixture fraction, is vitally important in studies of the reactive flow. The temperature marks the evolution of heat release and energy transfer, while species concentration gradients provide critical information on mixing and chemical reaction. Optical imaging techniques have the advantage of being non-invasive, which means that the studied process is not significantly perturbed by the measurement technique, and allowing for the acquisition of statistics in-situ. Spectroscopy offer intrinsic chemical specificity, in that different classes of molecules have specific spectral signatures serving as unique fingerprints for their identification. Laser-based diagnostics may in general provide measurements with exceptionally high spatial- and temporal resolution, which is important in producing reliable and accurate experimental data. Coherent anti-Stokes Raman spectroscopy (CARS) is one such versatile technique, which has had a profound impact on a wide variety of fields. It was pioneered in composition- and temperature measurements almost 40 years ago, and is referred to as authoritative with the level of accuracy and precision it may provide. A limitation still, has been its main applicability as a single point-measurement technique, where the experimenter needs to raster-scan the measurement samples assembling the spatial image. Because many complex systems can be fully characterized in multidimensional space, there is a large motivation for the advancement of multidimensional CARS imaging techniques.


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Soulcalibur: Broken Destiny is a fighting video game in the Soulcalibur series for PlayStation Portable (PSP). It uses many of the features of Soulcalibur IV, including its character customization mode. One of the goals of the game is "to target beginners and novice players with Soulcalibur IVs content." The game introduces two new characters to the series — Kratos from the God of War series and Dampierre, a new original character.

Characters

Soulcalibur: Broken Destiny includes 28 characters. All regular characters from Soulcalibur IV return, but the bonus characters and Star Wars guest characters from Soulcalibur IV are absent. At the 2009 Electronic Entertainment Expo, Namco Bandai revealed that they were to be replaced with a different guest character — Kratos from Sony's God of War series, and a new exclusive character named Dampierre.

Gameplay
The ability to change physical attributes and muscularity, which was present in Soulcalibur IV, has been removed. In this game, however, it has been replaced by the ability to adjust the rotation, position and size of some equipped items, such as certain headwear which might not fit exactly right, depending on what hairstyle is selected. Original characters can not be re-dressed with custom character items like in Soulcalibur IV, although their colors may be edited. In addition to editing the characters' appearance, the player may create a custom vs. screen photo for each custom character. The player has some control over the character's pose in the photo, as well as the camera position and the photo's frame and background. The number of custom character slots has been reduced from 50 in Soulcalibur IV to 16 in Broken Destiny.

Soulcalibur: Broken Destiny features an English language and Japanese language mode for both text and speech. Custom characters, however, can only be named using the ASCII naming interface, regardless of the language setting.

 Quick Match is a mode where players are given a list of A.I. players with their titles and stats and choose which opponent to fight. Players who have won against the A.I will receive "unique titles" that you can wear, the "title" is under their name during Versus Mode.
 The Gauntlet is the tutorial and story mode for Soulcalibur: Broken Destiny, which has similar story telling features like Soulcalibur II'''s Weapon Master mode, although the player is informed at the start that this story is non-canonical. Players are often given low health and required to guard or perform attacks within a short window of opening of the A.I. controlled tutor. There are altogether 34 chapters of tutorial in The Gauntlet, with two to four sub-missions in each of the stages. Players will need to clear all sub-missions with Rank A in order to unlock the next chapter. As a reward for completing The Gauntlet, players are given a new 'Broken Destiny' weapon for the character Siegfried.
 Trials consists of three portions — Trial of Attack, Trial of Defense, and Endless Trials. All of the portions are round-based battles. Trial of Attack rewards score according to how well a player can chain the attacks. Trial of Defense rewards score according to how well a player guard attacks and counterattacks. Endless Trials essentially combine both of the above trials and endless stages much like survival mode.
 Versus Mode is similar in features to Quick Match, except that the players are nearby PSP players who connect wirelessly through an ad hoc network.
 Training Mode which allows player to test out moves and practice in controlled condition, much the same as Practice Mode/Training in other previous Soulcalibur games.

Reception

The game received "favorable" reviews according to the review aggregation website Metacritic. Common criticisms were a lack of story, game modes, and online versus modes. In Japan, Famitsu'' gave it a score of one seven and three eights for a total of 31 out of 40.

References

External links

2009 video games
3D fighting games
Bandai Namco games
Crossover fighting games
God of War (franchise)
Ubisoft games
PlayStation Portable-only games
Soulcalibur series games
Fighting games
Video games set in the 17th century
Video game sequels
Multiplayer and single-player video games
PlayStation Portable games
Video games developed in Japan
Video games scored by Junichi Nakatsuru
Video games scored by Masaharu Iwata

redpajama_set_name: RedPajamaWikipedia


text:
import pytest

from awx.main.access import (
    WorkflowJobTemplateAccess,
    WorkflowJobTemplateNodeAccess,
    WorkflowJobAccess,
    # WorkflowJobNodeAccess
)

from awx.main.models import InventorySource, JobLaunchConfig

@pytest.fixture
def wfjt(workflow_job_template_factory, organization):
    objects = workflow_job_template_factory('test_workflow', organization=organization, persisted=True)
    return objects.workflow_job_template

@pytest.fixture
def wfjt_with_nodes(workflow_job_template_factory, organization, job_template):
    objects = workflow_job_template_factory(
        'test_workflow', organization=organization, workflow_job_template_nodes=[{'unified_job_template': job_template}], persisted=True)
    return objects.workflow_job_template

@pytest.fixture
def wfjt_node(wfjt_with_nodes):
    return wfjt_with_nodes.workflow_job_template_nodes.all()[0]

@pytest.fixture
def workflow_job(wfjt):
    return wfjt.workflow_jobs.create(name='test_workflow')

@pytest.mark.django_db
class TestWorkflowJobTemplateAccess:

    def test_random_user_no_edit(self, wfjt, rando):
        access = WorkflowJobTemplateAccess(rando)
        assert not access.can_change(wfjt, {'name': 'new name'})

    def test_org_admin_edit(self, wfjt, org_admin):
        access = WorkflowJobTemplateAccess(org_admin)
        assert access.can_change(wfjt, {'name': 'new name'})

    def test_org_admin_role_inheritance(self, wfjt, org_admin):
        assert org_admin in wfjt.admin_role
        assert org_admin in wfjt.execute_role
        assert org_admin in wfjt.read_role

    def test_org_workflow_admin_role_inheritance(self, wfjt, org_member):
        wfjt.organization.workflow_admin_role.members.add(org_member)

        assert org_member in wfjt.admin_role
        assert org_member in wfjt.execute_role
        assert org_member in wfjt.read_role

@pytest.mark.django_db
class TestWorkflowJobTemplateNodeAccess:

    def test_no_jt_access_to_edit(self, wfjt_node, org_admin):
        # without access to the related job template, admin to the WFJT can
        # not change the prompted parameters
        access = WorkflowJobTemplateNodeAccess(org_admin)
        assert not access.can_change(wfjt_node, {'job_type': 'check'})

    def test_node_edit_allowed(self, wfjt_node, org_admin):
        wfjt_node.unified_job_template.admin_role.members.add(org_admin)
        access = WorkflowJobTemplateNodeAccess(org_admin)
        assert access.can_change(wfjt_node, {'job_type': 'check'})

    def test_access_to_edit_non_JT(self, rando, workflow_job_template, organization, project):
        workflow_job_template.admin_role.members.add(rando)
        node = workflow_job_template.workflow_job_template_nodes.create(
            unified_job_template=project
        )
        assert not WorkflowJobTemplateNodeAccess(rando).can_change(node, {'limit': ''})

        project.update_role.members.add(rando)
        assert WorkflowJobTemplateNodeAccess(rando).can_change(node, {'limit': ''})

    def test_add_JT_no_start_perm(self, wfjt, job_template, rando):
        wfjt.admin_role.members.add(rando)
        access = WorkflowJobTemplateNodeAccess(rando)
        job_template.read_role.members.add(rando)
        assert not access.can_add({
            'workflow_job_template': wfjt,
            'unified_job_template': job_template})

    def test_add_node_with_minimum_permissions(self, wfjt, job_template, inventory, rando):
        wfjt.admin_role.members.add(rando)
        access = WorkflowJobTemplateNodeAccess(rando)
        job_template.execute_role.members.add(rando)
        inventory.use_role.members.add(rando)
        assert access.can_add({
            'workflow_job_template': wfjt,
            'inventory': inventory,
            'unified_job_template': job_template})

    def test_remove_unwanted_foreign_node(self, wfjt_node, job_template, rando):
        wfjt = wfjt_node.workflow_job_template
        wfjt.admin_role.members.add(rando)
        wfjt_node.unified_job_template = job_template
        access = WorkflowJobTemplateNodeAccess(rando)
        assert access.can_delete(wfjt_node)

@pytest.mark.django_db
class TestWorkflowJobAccess:

    @pytest.mark.parametrize("role_name", ["admin_role", "workflow_admin_role"])
    def test_org_admin_can_delete_workflow_job(self, role_name, workflow_job, org_member):
        role = getattr(workflow_job.workflow_job_template.organization, role_name)
        role.members.add(org_member)

        access = WorkflowJobAccess(org_member)
        assert access.can_delete(workflow_job)

    def test_wfjt_admin_can_delete_workflow_job(self, workflow_job, rando):
        workflow_job.workflow_job_template.admin_role.members.add(rando)
        access = WorkflowJobAccess(rando)
        assert not access.can_delete(workflow_job)

    def test_cancel_your_own_job(self, wfjt, workflow_job, rando):
        wfjt.execute_role.members.add(rando)
        workflow_job.created_by = rando
        workflow_job.save()
        access = WorkflowJobAccess(rando)
        assert access.can_cancel(workflow_job)

    def test_admin_cancel_access(self, wfjt, workflow_job, rando):
        wfjt.admin_role.members.add(rando)
        access = WorkflowJobAccess(rando)
        assert access.can_cancel(workflow_job)

    def test_execute_role_relaunch(self, wfjt, workflow_job, rando):
        wfjt.execute_role.members.add(rando)
        JobLaunchConfig.objects.create(job=workflow_job)
        assert WorkflowJobAccess(rando).can_start(workflow_job)

    def test_cannot_relaunch_friends_job(self, wfjt, rando, alice):
        workflow_job = wfjt.workflow_jobs.create(name='foo', created_by=alice)
        JobLaunchConfig.objects.create(
            job=workflow_job,
            extra_data={'foo': 'fooforyou'}
        )
        wfjt.execute_role.members.add(alice)
        assert not WorkflowJobAccess(rando).can_start(workflow_job)

    def test_relaunch_inventory_access(self, workflow_job, inventory, rando):
        wfjt = workflow_job.workflow_job_template
        wfjt.execute_role.members.add(rando)
        assert rando in wfjt.execute_role
        workflow_job.created_by = rando
        workflow_job.inventory = inventory
        workflow_job.save()
        wfjt.ask_inventory_on_launch = True
        wfjt.save()
        JobLaunchConfig.objects.create(job=workflow_job, inventory=inventory)
        assert not WorkflowJobAccess(rando).can_start(workflow_job)
        inventory.use_role.members.add(rando)
        assert WorkflowJobAccess(rando).can_start(workflow_job)

@pytest.mark.django_db
class TestWFJTCopyAccess:

    def test_copy_permissions_org_admin(self, wfjt, org_admin, org_member):
        admin_access = WorkflowJobTemplateAccess(org_admin)
        assert admin_access.can_copy(wfjt)

        wfjt.organization.workflow_admin_role.members.add(org_member)
        admin_access = WorkflowJobTemplateAccess(org_member)
        assert admin_access.can_copy(wfjt)

    def test_copy_permissions_user(self, wfjt, org_admin, org_member):
        '''
        Only org admins and org workflow admins are able to add WFJTs, only org admins
        are able to copy them
        '''
        wfjt.admin_role.members.add(org_member)
        member_access = WorkflowJobTemplateAccess(org_member)
        assert not member_access.can_copy(wfjt)

    def test_workflow_copy_warnings_inv(self, wfjt, rando, inventory):
        '''
        The user `rando` does not have access to the prompted inventory in a
        node inside the workflow - test surfacing this information
        '''
        wfjt.workflow_job_template_nodes.create(inventory=inventory)
        access = WorkflowJobTemplateAccess(rando, save_messages=True)
        assert not access.can_copy(wfjt)
        warnings = access.messages
        assert 'inventories_unable_to_copy' in warnings

    def test_workflow_copy_no_start(self, wfjt, inventory, admin_user):
        # Test that un-startable resource doesn't block copy
        inv_src = InventorySource.objects.create(
            inventory = inventory,
            source = 'custom',
            source_script = None
        )
        assert not inv_src.can_update
        wfjt.workflow_job_template_nodes.create(unified_job_template=inv_src)
        access = WorkflowJobTemplateAccess(admin_user, save_messages=True)
        access.can_copy(wfjt)
        assert not access.messages

    def test_workflow_copy_warnings_jt(self, wfjt, rando, job_template):
        wfjt.workflow_job_template_nodes.create(unified_job_template=job_template)
        access = WorkflowJobTemplateAccess(rando, save_messages=True)
        assert not access.can_copy(wfjt)
        warnings = access.messages
        assert 'templates_unable_to_copy' in warnings

redpajama_set_name: RedPajamaGithub


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text:
Health & Science / Conception
BPA & Phthalates: The Reason Why Plastic is Dangerous for Children
The common chemicals BPA and phthalates are found nearly everywhere from food containers to toys, and side effects from exposure can have lifelong consequences.
By Matthew Utley
Updated Oct 14 2019, 4:51 PM
BPA-free is often the selling point for baby products, but there's rarely an explanation about what BPA is and why BPA plastics should be avoided. Essentially BPA and phthalates – a related class of chemicals – are plasticizers that can be found in food packaging, steel can liners, drug coatings, and cosmetics. They are commonly used to make plastic strong lightweight, and flexible. And considering bps effects, there are some very good reasons they should be avoided.
BPA Effects on Children
BPA exposure can have serious consequences prenatal development, sexual maturity, and reproduction. And despite a trend of childcare products ranging from dining wear to car seat accessories and bath toys being specifically engineered to be BPA free, BPA is everywhere. That's despite the American Association of Pediatrics warning against this use of BPA in baby products. Even trace amounts of BPA can be a problem, because of the way they interact with the body's normal production of hormones.
"BPA's should be our biggest concern because they contribute to the overall toxicity in our environment," warns Mark Menolascino, MD, medical director of the Meno Clinic Center for Functional Medicine in Jackson Hole, WY, and Chief Medical Officer for Genexa."BPAs create hormone disruption in multiple forms, including thyroid and adrenal, as well as the female and male hormones we're familiar with," says Menolascino.
What are BPA and Phthalates?
They are practically ubiquitous: BPA and phthalates appear in plastics, canned food, cosmetics, and even credit card receipts.
They are hormone disruptors: BPA and phthalates interfere with the body's normal use of hormones; not just estrogen and testosterone, but thyroid and adrenal hormone production as well.
They can cause physical changes: they can lead to genital changes that can cause lower sperm count in boys.
They can cause systemic changes: they can severely disrupt hormone regulation in girls and can even cause early menopause.
How stressful do you find days when your kids can't go outside?
Not stressful at all. We make the best of it
Sometimes they are, sometimes they aren't. Depends on the circumstance.
Extremely stressful. It's hard to entertain them
BPA Effects in Pregnant Women
In pregnant women, hormones are very much necessary for prenatal development. It's hormones like estrogen and testosterone that cue the development of a baby's genitals. In the case of phthalates, this can manifest itself in changes in the physical structures of the genitals. A 2014 Swedish study suggests that phthalate exposure shortened the anogenital distance (ADG) in Swedish boys. ADG is the measurement between the anus and the genitals. The shorter the distance between the two, the greater the risks of problems including low sperm count and undescended testicles. Prenatal and early exposure to BPA and phthalates can affect a child throughout their entire life. The repercussions can manifest in any number of ways, regardless of sex.
"In boys, BPAs can affect their ability for optimal performance, including testosterone levels, and may even affect fertility. If you want your son to have optimal performance, whether that be as someone who excels in math or someone who excels in sports, toxicity from chemicals like BPA's can sabotage that performance," says Menolascino. "In women, exposure to BPA can cause PMS, early menopause, acne, and is a strong source of other hormone disruptors. Any hormone disruptor, including BPAs, can change the hormone pathway, and other phthalates can have similar effects and increase our overall toxic load."
BPA use is reduced in the production of certain plastics, and phthalates are currently being regulated out of toys, but there are still no real guidelines about safe levels of phthalates in pregnant women. Because of this, many endocrine specialists have recommended that pregnant women stay away from processed food (which is often in contact with plastic) and keep a close eye on the ingredients of their cosmetics. Stainless steel and glass dishes are safer options for heating food (although not stainless steel in the microwave, obviously.) But even with those precautions, BPA and phthalates are found nearly everywhere.
"The biggest concern for all of us and our kids today," says Menolascino, "is the toxicity in our environment."
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The Bill Gates Code hits Netflix in September and already has a trailer | Culture
Favorite animal? Dog. Favorite food? Hamburger. What do you eat for breakfast? Nothing. You would expect these responses from anyone, probably a child, but they came from one of the richest men in the world: this is how the trailer for The Bill Gates Code, a three-part documentary that premieres at Netflix on September 20.
Portraying on the screen the life of one of the most iconic names in the world of business and philanthropy is a huge challenge. It is not because of Bill Gates' visionary personality, but because of the need not to succumb to the expectations that such a biographical production can generate.
It is to be expected that many people watch the documentary in order, albeit discreetly, to find clues on how to achieve success. That expectation must be met, but the trailer implies that, for Gates, success is not necessarily about winning big fortunes.
In fact, the documentary should reveal, in part, how Bill Gates managed to reinvent himself. When I was in charge of Microsoft, Gates was often seen as a greedy person, able to make cunning decisions so as not to be at a disadvantage.
Perhaps a moment that faithfully portrays this phase was the Gates' testimony in August 1998 about the antitrust lawsuit that Microsoft was facing. Even though he showed nervousness, the businessman thought well to avoid that his answers were used against him and did not hesitate to say "I don't know" or "I don't remember" when he was trapped.
Since leaving Microsoft's leadership, however, Bill Gates has mastered the image of a generous philanthropist, who tries to improve the world by supporting social projects and research to eradicate diseases and renewable energy, for example.
Apparently, it is exactly this change that The Bill Gates Code will show, but still shedding light on other important moments in the life of the Microsoft co-founder.
On twitter, Gates has already left his invitation to the premiere: "I spent the last few years participating in a Netflix series that portrays my work and my life, which will be released on September 20th. I hope you enjoy the work they did ".
By FaceApp, Procon-SP fines Google and Apple up to R $ 10 million – Brazil
Windows 10 changes tablet experience to two-in-one PCs | Applications and Software


After nine years of diligent training, Weed High School student Griffin Hough realized a long-held dream and took his first degree black belt exam in early December.
The black belt exam is the time to "Put Up or Shut Up." With a panel of six black belt instructors watching every move, there's no place to hide. Candidates must demonstrate exciting physical skills along with precision and accuracy.
Hough traveled to UC Berkeley with his parents and demonstrated a variety of skills including basic movements – blocks, strikes, stances; forms (prearranged movements); advanced techniques requiring jumping and spinning; and full-contact free sparring in front of over 100 spectators and martial artists.
In addition to the physical exam, Hough was required to submit a personal essay and book report to further expand his understanding of, and appreciation for, the martial arts and how they benefit other areas of his life.
Family support was also a key ingredient to his accomplishment.
Weed High School English teacher Mary Jo Coffman sees the results firsthand in Hough's performance at school.
Despite his skills, Hough is still a typical teenager in many ways.
MSMAP holds its Winter Taekwondo Promotion Exam on Wednesday, February 9 at 5pm at 612-A S. Mt. Shasta Blvd. Contact them at 859-2024, www.MtShastaMA.org, and on Facebook.
Go to: What is Taekwondo?
Vitaliy shares what it takes to get a black belt: it's not easy. It takes lots and lots of hard work...and plenty of support from friends and family.
23 children, teens, and adults met the challenge for their next rank — along with an added test of their judgement when facing the unexpected.
15 children took up the challenge to test for their next rank: beginning, intermediate, & advanced.
MSMAP holds its largest promotion exam and challenges 27 people to show what they know and do their best.
A comprehensive article on the positive results of martial arts training — in the studio, in school, in life.
The impressions of several families about one of the largest promotion exams in MSMAP history.
MSMAP hosted over 100 participants at McCloud High School for an incredible international martial arts summer camp in August 2006. Master instructors and Olympic medalists led training in taekwondo, yongmudo, judo, kumdo, and ssirum (Korean traditional wrestling).