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Abstract
In this study, the effects of edible bird’s nest on sexual function of male castrated rats were investigated for the first time. The testosterone (T), luteinizing hormone (LH) and estradiol (E2) levels, the penis and prostate and seminal vesicle indexes, and the protein expression of endothelial nitric oxide synthase (eNOS) were estimated. The prostate and seminal vesicle index and the expression of eNOS increased significantly in groups treated with edible bird’s nest in comparison to the control group. The results demonstrated that the edible bird’s nest may promote the sexual function of male castrated rats and T was supposed to be responsible for this function. Edible bird’s nest may serve as an effective medicine for erectile dysfunction (ED) treatment.
Key words: Edible bird’s nest, sexual function, castrated rats, erectile dysfunction, hormones, nitric oxide synthase.
... Firstly, EBN was reported to have a positive proliferative effect in in vitro study (Aswir and Wan Nazaimoon, 2011;Abidin et al., 2011;Roh et al., 2012). Second, EBN was also reported containing a few male reproductive hormones such as FSH, LH, and testosterone (Ma and Liu, 2012a) and showed potential as an alternative treatment for erectile dysfunction (Ma et al., 2012). These EBN characteristics align with the current study objective and therefore becomes the reason for this selection interest. ...
Children are vulnerable to the radiofrequency radiation (RFR) emitted by Wi-Fi devices. Nevertheless, the severity of the Wi-Fi effect on their reproductive development has been sparsely available. Therefore, this study was conducted to evaluate the Wi-Fi exposure on spermatogonia proliferation in the testis. This study also incorporated an approach to attenuate the effect of Wi-Fi by giving concurrent edible bird’s nest (EBN) supplementation. It was predicted that Wi-Fi exposure reduces spermatogonia proliferation while EBN supplementation protects against it. A total of 30 (N = 30) 3-week-old Sprague Dawley weanlings were divided equally into five groups; Control, Control EBN, Wi-Fi, Sham Wi-Fi, and Wi-Fi + EBN. 2.45 GHz Wi-Fi exposure and 250 mg/kg EBN supplementation were conducted for 14 weeks. Findings showed that the Wi-Fi group had decreased in spermatogonia mitosis status. However, the mRNA and protein expression of c-Kit-SCF showed no significant decrease. Instead, the reproductive hormone showed a reduction in FSH and LH serum levels. Of these, LH serum level was decreased significantly in the Wi-Fi group. Otherwise, supplementing the Wi-Fi + EBN group with 250 mg/kg EBN resulted in a significant increase in spermatogonia mitotic status. Even though EBN supplementation improved c-Kit-SCF mRNA and protein expression, the effects were insignificant. The improvement of spermatogonia mitosis appeared to be associated with a significant increase in blood FSH levels following EBN supplementation. In conclusion, the long-term Wi-Fi exposure from pre-pubertal to adult age reduces spermatogonia proliferation in the testis. On the other hand, EBN supplementation protects spermatogonia proliferation against Wi-Fi exposure.
... Many studies have reported the medicinal and therapeutic effects in EBN, such as the ability in rejuvenating skin, inhibiting influenza virus and inflammation, proliferating cell, enhancing bone strength and dermal thickness, reducing tumor production, treating erectile dysfunction and osteoporosis. [9][10][11][12][13][14][15] Owing to its potential abilities and high price, EBN is increasingly fraud substituted with lower quality EBN and adulterated with cheaper ingredients such as Tremella fungus, karaya gum, red seaweed and fried porcine skin, and then marketed at a premium price for greater financial gain. [16] Visual inspection is less likely to detect these fraud substitutions and adulterants in EBN because they have similar appearance with the genuine EBN. ...
Authenticity of food is of great importance to ensure food safety and quality, and to protect consumer rights. A rapid and accurate method for authentication of edible bird’s nest (EBN) was proposed by using nutritional profile and chemical composition, and pattern recognition analysis. The authentication of EBN includes identification and classification of EBN by production origin (houses or caves), species origin (Aerodramus fuciphagus or Aerodramus maximus) and geographical origin (Peninsular Malaysia or East Malaysia) based on their active compositional content. Three pattern recognition methods, principal component analysis (PCA), hierarchical cluster analysis (HCA) and linear discriminant analysis (LDA), were employed to develop classification models for authentication of EBN origins. Compared to PCA and HCA, LDA is more accurate and efficient in distinguishing EBN by different production, species, and geographical origins, having classification ability of 100% and prediction ability of 92% as validated by cross-validation method. The key chemical markers for production origin differentiation are total phenolic content, zinc, valine, and calcium, while for species origin discrimination are sialic acid, serine, phenylalanine and valine, and for geographical origin differentiation are arsenic and mercury. The findings suggest that nutritional and chemical profiles combined with pattern recognition analysis are promising strategy for rapid authentication of EBN and its products.
... EBN consumption has been initiated since ancient times, specifically among global Chinese communities, in the belief that EBN can maintain, enhance and boost their health. Many functional effects of EBN such as anti-inflammatory, antiaging, anti-influenza virus infection, stimulating cell proliferation, improving bone strength and dermal thickness, promoting hair growth, and treating erectile dysfunction and osteoarthritis have been reported in vivo and in vitro Guo et al., 2006;Ma, Liu, & Dai, 2012;Matsukawa et al., 2011;Okajima, 2009;Rashed & Nazaimoon, 2010). These beneficial effects greatly depend on the quality of EBN that influenced by the nesting season and harvesting period, and the habitat conditions, food availability and species origin of swiftlets. ...
... It is traditionally regarded as a functional food for skin rejuvenating and health promoting. EBN has been demonstrated scientifically to inhibit inflammation and influenza virus, proliferate cell, improve bone strength and dermal thickness, reduce tumour production, and treat erectile dysfunction and osteoporosis (Chua et al., 2013;Guo et al., 2006;Kong et al., 1987;Ma, Liu, & Dai, 2012;Matsukawa et al., 2011;Roh et al., 2011;Vimala, Hussain, & Nazaimoon, 2012). EBN is produced from the saliva of swiftlet species, which belongs to the family Apodidae and genus Aerodramus. ...
The increasing demand and consumption of edible bird's nest (EBN) by people worldwide has contributed to the food fraud issue. To ensure the authenticity of EBN in regard to their origin, rapid and accurate analytical methods are very much needed. In this study, forensically informative nucleotide sequencing (FINS) technique based on mitochondrial and nuclear DNA sequences, and phylogenetic analysis was performed to identify the species and production origins of raw and commercial EBNs. The cytochrome b (Cyt b), NADH dehydrogenase subunit 2 (ND2), 12S ribosomal RNA and beta-fibrinogen intron 7 gene markers used were able to identify and classify EBN produced by Aerodramus fuciphagus and Aerodramus maximus. It was newly discovered that EBN from man-made houses and natural caves were genetically differentiable using the mitochondrial Cyt b and ND2 genes. The phylogenetic results revealed that all EBN samples were well-separated into two groups following their species origin and production origin. A rapid and cost-effective identification alternative of SYBR green I based real-time PCR assay targeting a 177 bp of the mitochondrial Cyt b gene was developed and it efficiently differentiated genuine EBN from counterfeits. This FINS and SYBR green I based real-time PCR are highly sensitive, specific and reliable methods for identification of EBN origins and could be useful for preventing fraud substitution and mislabelling of EBN to ensure food safety.
... All these studies confirmed the previous study which reported that EBN contains epidermal growth factor-like substances which could help to trigger cell division that results in skin rejuvenation (Kong et al., 1987). Apart from this, EBN extract was found to exhibit versatile medical potentials such as enhancing bone strength and dermal thickness (Matsukawa et al., 2011), treating erectile dysfunction (Ma, Liu, & Dai, 2012) as well as serving as an alternative chondro-protective agent in curing osteoarthritis (Chua et al., 2013). ...
Edible bird's nest (EBN) swiftlet existed naturally 48,000 years ago in caves as their natural dwellings. Nowadays, edible bird's nest has become a very important industry due to its high nutritional, medicinal and economic value. Additionally, edible bird's nest has a long quality guarantee period. Obviously, the nutritional components and medicinal functions vary depending on geographical origins. Recently, the global demand for edible bird's nest has markedly increased, accompanied by the increasing attention of all key players of the global food trade system, i.e., producers, consumers, traders and the authorities to obtain safe and high-quality edible bird's nest. Hence, this target can be accomplished via the enforcement of an efficient and universal geo-tracing technique. Current methods of the geo-tracking of edible bird's nest, i.e., automation, physical and analytical techniques have several limitations and all of them fail to discriminate different quality grades of edible bird's nest. Meanwhile, in many studies and applications, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) has proven to be a “cutting edge” technique for greatly enhance food traceability from field to fork through its ability in distinguishing the food products in terms of their quality and safety. This article provides an overview of (1) edible bird's nest as a multiuse strategic food product, (2) quality issues associated with edible bird’s nest including implications that the site of acquisition of the edible bird’s nest has food safety implications, (3) current regulations and geo-tracking approaches to ensure the safety and quality of edible bird’s nest with the special focus on polymerase chain reaction-denaturing gradient gel electrophoresis technique as a vigorous and universal geo-tracing tool to be suggested for edible bird's nest geo-traceability.
Edible Bird’s Nest (EBN) is the most prized health delicacy among the Chinese population in the world. Although some scientific characterization and its bioactivities have been studied and researched, no lights have been shed on its actual composition or mechanism. The aim of this review paper is to address the advances of EBN as a therapeutic animal bioproduct, challenges and future perspectives of research involving EBN. The methodology of this review primarily involved a thorough search from the literature undertaken on Web of Science (WoS) using the keyword “edible bird nest”. Other information were obtained from the field/market in Malaysia, one of the largest EBN-producing countries. This article collects and describes the publications related to EBN and its therapeutic with diverse functional values. EBN extracts display anti-aging effects, inhibition of influenza virus infection, alternative traditional medicine in athletes and cancer patients, corneal wound healing effects, stimulation of proliferation of human adipose-derived stem cells, potentiate of mitogenic response, epidermal growth factor-like activities, enhancement of bone strength and dermal thickness, eye care, neuroprotective and antioxidant effects. In-depth literature study based on scientific findings were carried out on EBN and its properties. More importantly, the future direction of EBN in research and development as health-promoting ingredients in food and the potential treatment of certain diseases have been outlined.
Edible bird’s nest (EBN) is reported to have a positive in vitro proliferative effect and contain male reproductive hormones. Spermatogonia cells proliferate during spermatogenesis under male reproductive hormones stimulation that include testosterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH). Characterization of EBN through liquid chromatography-mass spectrometry (LCMS) has found testosterone as a base peak. Six types of amino acids, estradiol and sialic acid were among the major peaks that have been characterized. Based on the presence of these reproductive components, this study evaluated different doses of EBN on sperm parameters and male reproductive hormones of Sprague Dawley rats. Sixteen Sprague Dawley rats at the age of eight weeks were randomly and equally divided into four groups, which are Control, 10 mg/kg BW/d 50 mg/kg BW/d, and 250 mg/kg BW/d EBN group. The rats were fed with EBN enriched pellet daily and water ad-libitum. Rats were sacrificed and the organ was weighed for organ coefficients after eight weeks of treatment. Sperm concentration, percentage of sperm motility, and sperm viability were evaluated. Meanwhile, ELISA method was used to measure testosterone, FSH, and LH. Findings showed that there were no significant differences in organ coefficient between groups. Supplementation of 250 mg/kg BW/d EBN demonstrated a significant increase in sperm concentration, percentage of sperm motility as well as FSH and LH level compared to 10 mg/kg BW/d group. There was a dose-dependent increase in testosterone level but was not significant between groups. Based on these findings, EBN is concluded to have crucial effects on male reproductive parameters.
The high priced cave edible bird's nest (EBN) has attracted unscrupulous EBN producers to adulterate EBN with lower priced house-farmed EBN due to the fact that both are almost indistinguishable by visual inspection. In the present study, major mineral contents such as calcium, sodium, magnesium and potassium of both EBN types were analysed using inductively coupled plasma-optical emission spectrometry (ICP-OES). Three pattern recognition techniques namely hierarchical cluster analysis (HCA), principal component analysis (PCA) and linear discriminant analysis (LDA) were employed to determine the influence of harvesting origins on mineral profiles. With the use of HCA and PCA, EBN samples have successfully been grouped into two distinct clusters. From the PCA score plot, principal component 1 (49.53 %) and principal component 2 (41.11%) accounted for 90.64% of the total variability. In addition, LDA presented excellent performance in discriminating and predicting membership of the two EBN sample types with classification rate of 100%.
To investigate the relationship between oxytocin (OT) and male infertility, serum OT baseline concentration and oxytocin receptor (OTR) gene expression in fertile and infertile men were investigated.
Twenty obstructive azoospermia patients, twenty five idiopathic asthenozoospermia patients, twenty idiopathic oligozoospermia patients and twenty healthy subjects were taken into consideration. Serum OT baseline concentration was determined by radioimmunoassay. Moreover, serum concentration of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone (T) were determined by chemoluminescence to evaluate the correlation with OT. OTR gene promotor and OTR mRNA expressions were determined by polymerase chain reaction and reverse transcriptase-polymerase chain reaction, respectively. OTR protein expression was also performed by Western Blot.
Serum OT baseline concentrations in infertile groups were significantly higher than in fertile group (F0.05/2(2,82) = 8.29, p < 0.001). Serum baseline concentration of OT was not correlated with that of LH, FSH and T. There was no significant difference in gene sequences of OTR gene promotor and OTR mRNA when comparing infertile patients with fertile. Human OTR was in the form of oligomers and monomers, and the oligomers were in the majority containing tetramers and hexamers. Monomer expression was significantly higher in idiopathic asthenozoospermia and idiopathic oligozoospermia than that in obstructive azoospermia and control group (F0.05/2(2,82) = 115.50, p < 0.001). There was no significant difference in oligomer expression between different groups, but 20% of idiopathic asthenozoospermia cases showed a decrease.
Significantly different OT baseline concentrations and OTR expressions between fertile and infertile men strongly suggest that OT/OTR system is likely to be linked with male infertility, providing new insights into the pathogenesis and treatment of male infertility.
The most accurate method of determining protein concentration is probably acid hydrolysis followed by amino acid analysis. Most other methods are sensitive to the amino acid composition of the protein and absolute concentrations cannot be obtained. The procedure of Lowry et al. (1) is no exception, but its sensitivity is moderately constant from protein to protein, and it has been so widely used that Lowry protein estimations are a completely acceptable alternative to a rigorous absolute determination in almost all circumstances where protein mixtures or crude extracts are involved.
Erectile dysfunction (ED) is a common male sexual disorder. The relative benefits and harms of pharmacologic therapies for ED, as well as the value of hormonal testing in men with ED, are uncertain.
To evaluate the efficacy and harms of oral phosphodiesterase-5 (PDE-5) inhibitors and hormonal treatments for ED and assess the effect of measuring serum hormone levels on treatment outcomes for ED.
English-language studies from MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, PsycINFO, AMED, and SCOPUS through April 2009. Trial reference lists also were scanned.
Randomized, controlled trials (RCTs) of oral PDE-5 inhibitors and hormonal treatment for ED, and observational studies reporting measurement of serum hormone levels, prevalence of hormonal abnormalities, or both in men with ED.
Two independent reviewers abstracted data on study, participant, and treatment characteristics; efficacy and harms outcomes; and prevalence of hormonal abnormalities.
Data, primarily from short-term trials (<or=12 weeks), indicate that PDE-5 inhibitors were more effective than placebo in improving sexual intercourse success (69.0% vs. 35.0%). The proportion of men with improved erections was significantly greater among those treated with PDE-5 inhibitors (range, 67.0% to 89.0%) than with placebo (range, 27.0% to 35.0%). The PDE-5 inhibitors were associated with increased risk for any adverse events compared with placebo (for example, relative risk with sildenafil, 1.72 [95% CI, 1.53 to 1.93]). In 4 head-to-head RCTs comparing sildenafil, vardenafil, and tadalafil, improvement of ED and adverse events did not differ among treatments. Results from 15 RCTs evaluating hormonal treatment of ED were inconsistent on whether treatment improved outcomes. Evidence was insufficient regarding whether men with ED had a higher prevalence of hypogonadism than men without ED.
Many RCTs were of low methodological and reporting quality, particularly those involving hormonal treatments or directly comparing different PDE-5 inhibitors. Most RCTs provided only short-term efficacy and harms data.
Oral PDE-5 inhibitors improved erectile functioning and had similar efficacy and safety profiles. Results on the efficacy of hormonal treatments and the value of hormone testing in men with ED were inconclusive.
Agency for Healthcare Research and Quality.
Efficient production of high quality semen is of major importance to artificial insemination (AI) organizations. The semen produced should be free of contagious organisms, be of high quality, have good storage properties, fertilizing capacity and be of high genetic value. The best approach to prevent the spreading of microorganisms via semen in the process of AI is to collect semen from boars free from specific diseases, for example pseudorabies virus or leptospirosis. Antibiotics are added to the semen to suppress proliferation of microorganisms or even reduce their number. Sperm production is influenced by many factors such as season, collection frequency, breed and age. The average number of sperm cells produced per boar per week can vary more than 30% within one AI station, depending on the breed. Boar selection and boar management markedly influence the efficiency of sperm production. Sperm quality should be evaluated by fertility results obtained at breeding farms related to both farrowing rate and litter size to ensure a good quality monitoring system. A quality control system should be established to provide maximum reliability to customers.
A few species of swiflets (genus Aerodramus) build edible nests that are consumed by humans worldwide, as a delicacy known as the “Caviar of the East” or as a medicinal food. This study reports on the compositional properties of two types of nest, the white nest and the red “blood” nest. The order of composition (from lowest to highest) was found to be identical for both types of nests, i.e., lipid (0.14–1.28%), ash (2.1%), carbohydrate (25.62–27.26%) and protein (62–63%). It was also found that both nests share a common 77 KDa protein that has properties similar to those of the ovotransferrin protein in eggs. This protein may be partially responsible for the severe allergic reactions that sometimes occur among young children who consume edible bird’s nest products. It was found that SDS–PAGE electrophoretic fingerprinting might serve as a useful analytical technique for differentiating between white and red nests and for determining if the more expensive “blood” nest was adulterated with the less expensive white nest. Also evaluated were different analytical methodologies for detecting adulterants. Three of the most common adulterants found in retail bird’s nests are karaya gum, red seaweed, and tremella fungus, and they are routinely incorporated during commercial processing prior to final sale. Using crude protein determination, it was found that these adulterants (which typically accounted for 2–10% of the finished nest), reduce the overall crude protein content of the genuine white bird’s nest by as much as 1.1–6.2%. A modified xanthoproteic nitric acid test for proteins proved to be a rapid, and simple test to detect adulteration in both whole and finely ground nests, and would be suitable in the field where analytical facilities are not readily available.
Erectile dysfunction (ED) is a clinical disorder that results from a continuous spectrum of clinical factors, including physical illness (comprising the organic component of ED), reaction to stress (the intrapsychic component of ED) and relationship difficulties (the relationship component of ED). Testosterone clearly has a relevant role in all three causes of ED; the usefulness of this hormone in the treatment of ED has not, however, been completely clarified. The main physiological action of testosterone in the male sexual response is to regulate the timing of the erectile process as a function of sexual desire, thereby coordinating penile erection with sex. The link between ED, hypogonadism and underlying disorders (such as metabolic syndrome and type 2 diabetes mellitus) is nowadays well documented. The recognition of underlying disorders might be useful in motivating men with ED to improve their health-related lifestyle choices. Hence, patients with ED might be considered 'lucky', because their disorder offers the opportunity to undergo medical examinations to detect underlying disease. Both ED and hypogonadism are treatable conditions. A range of testosterone preparations are available for supplementation; their combination with phosphodiesterase 5 inhibitors might improve outcomes in some cases.
Nitric oxide has been identified as an endothelium-derived relaxing factor in blood vessels. We tried to determine whether it is involved in the relaxation of the corpus cavernosum that allows penile erection. The relaxation of this smooth muscle is known to occur in response to stimulation by nonadrenergic, noncholinergic neurons.
We studied strips of corpus cavernosum tissue obtained from 21 men in whom penile prostheses were inserted because of impotence. The mounted smooth-muscle specimens were pretreated with guanethidine and atropine and submaximally contracted with phenylephrine. We then studied the smooth-muscle relaxant responses to stimulation by an electrical field and to nitric oxide.
Electrical-field stimulation caused a marked, transient, frequency-dependent relaxation of the corpus cavernosum that was inhibited in the presence of N-nitro-L-arginine and N-amino-L-arginine, which selectively inhibit the biosynthesis of nitric oxide from L-arginine. The addition of excess L-arginine, but not D-arginine, largely reversed these inhibitory effects. The specific liberation of nitric oxide (by S-nitroso-N-acetylpenicillamine) caused rapid, complete, and concentration-dependent relaxation of the corpus cavernosum. The relaxation caused by either electrical stimulation or nitric oxide was enhanced by a selective inhibitor of cyclic guanosine monophosphate (GMP) phosphodiesterase (M&B 22,948). Relaxation was inhibited by methylene blue, which inhibits cyclic GMP synthesis.
Our findings support the hypothesis that nitric oxide is involved in the nonadrenergic, noncholinergic neurotransmission that leads to the smooth-muscle relaxation in the corpus cavernosum that permits penile erection. Defects in this pathway may cause some forms of impotence.
Nitric oxide (NO) is a cytotoxic agent of macrophages, a messenger molecule of neurons, and a vasodilator produced by endothelial cells. NO synthase, the synthetic enzyme for NO, was localized to rat penile neurons innervating the corpora cavernosa and to neuronal plexuses in the adventitial layer of penile arteries. Small doses of NO synthase inhibitors abolished electrophysiologically induced penile erections. These results establish NO as a physiologic mediator of erectile function.
Nitric oxide (NO), an active free radical formed during the conversion of arginine to citrulline by the enzyme NO synthase (NOS), mediates vasorelaxation, cytotoxicity, and neurotransmission. Neurons containing NOS (NOergic) are located in the hypothalamus. These NOergic neurons control the release of several hypothalamic peptides. Release of NO from these NOergic neurons stimulates pulsatile release of luteinizing hormone-releasing hormone (LHRH) in vivo and LHRH release in vitro. LHRH not only induces LH release, which induces ovulation, but also facilitates female sexual behavior. Sexual behavior can be induced reliably in estrogen-primed ovariectomized female rats by progesterone (P). This behavior consists of proceptive behavior to attract the male and the assumption of a clear characteristic posture, lordosis, when mounted by the male. To ascertain the role of NO in the control of sexual behavior in female rats, an inhibitor of NOS, NG-monomethyl-L-arginine was microinjected into the third cerebral ventricle (3V) of conscious, ovariectomized, estrogen-primed rats with indwelling cannulae. NG-Monomethyl-L-arginine (10-1000 micrograms) prevented P-facilitated lordosis when administered intracerebroventricularly into the 3V, 20 min prior to the 3V injection of P. NG-Monomethyl-D-arginine, which does not inhibit NOS, did not inhibit lordosis under the same experimental conditions. Microinjection into the 3V of sodium nitroprusside (SNP), which spontaneously releases NO, facilitated lordosis in estrogen-primed rats in the absence of P. The facilitation of lordosis induced by either P or SNP was prevented by intracerebroventricular injection of hemoglobin, which binds NO. Lordosis facilitated by P or SNP was blocked by injection of LHRH antiserum into the 3V. The results are interpreted to mean that the P-facilitated lordosis response is mediated by LHRH release. Furthermore, since NO release from SNP also facilitates lordosis in the absence of P and this response could be blocked by LHRH antiserum, we conclude that P brings about the release of NO, which stimulates LHRH release that facilitates lordosis. Thus, the results indicate that NO induces LHRH release and that LHRH then plays a crucial role in mediation of sexual behavior in the female rats.
Androgens are essential for the expression of normal libido in the male, but their role in the maintenance of the erectile response in humans is controversial. It has been shown previously in the rat that castration induces 1) loss of penile reflexes; and 2) considerable reduction in the erectile response to electric field stimulation (EFS) of the cavernosal nerve. Both of these effects can be reversed by testosterone replacement. The current study was performed to determine whether these testosterone effects are mediated via its conversion to dihydrotestosterone (DHT), and to what extent the synthesis of the mediator of penile erection, nitric oxide, is affected by castration and androgen replacement. Five-month-old rats were either castrated or left intact. The orchiectomized rats were implanted with SILASTIC brand silicon tubing (Dow Corning) containing testosterone or DHT with or without daily injections of the 5 alpha-reductase inhibitor finasteride. After 7 days, rats were submitted to EFS and the intracavernosal pressure was recorded. Castration reduced the EFS-induced erectile response by 50% in comparison with intact rats and testosterone restored this decrease to normal. When finasteride was given to these testosterone-treated castrate rats, erectile response was not restored. DHT was as effective as testosterone in restoring response to EFS in castrates and this effect was not decreased by finasteride. Nitric oxide synthase activity in the penile cytosol was measured by the arginine-citrulline conversion and was found to correlate with the EFS determinations. These results show that DHT is the active androgen in the prevention of erectile failure seen in castrated rats, and suggest that this effect may be mediated, at least partially, by changes in nitric oxide synthase levels in the penis.
Nitric oxide (NO) synthase, the enzyme which converts arginine into citrulline plus NO, a highly active free radical, has been found in many neurons in the brain, including neurons in the hypothalamus. Our previous experiments showed that norepinephrine-induced prostaglandin E2 release from hypothalamic explants incubated in vitro is mediated by NO. Since the release of luteinizing hormone-releasing hormone (LHRH) is also driven by norepinephrine and prostaglandin E2, we hypothesized that NO might also control pulsatile release of LHRH in vivo, resulting in turn in pulsatile release of luteinizing hormone (LH). To ascertain the role of NO in control of pulsatile LH release in vivo, an inhibitor of NO synthase, NG-monomethyl-L-arginine (NMMA), was microinjected into the third cerebral ventricle (1 mg/5 microliters) of conscious castrate male rats at time 0 and 60 min later; blood samples were taken every 10 min during this period. NMMA blocked pulsatile LH release within 20 min, and plasma LH concentration declined further without pulses after the injection at 60 min. Pulsatile release of LH was not altered in diluent-injected controls. NMMA did not alter pulsatile release of follicle-stimulating hormone, which suggests that its release does not require NO. Incubation of medial basal hypothalami with norepinephrine (10 microM) induced an increase in LHRH release that was inhibited by NMMA (300 microM). NMMA alone did not alter basal LHRH release, whereas it was augmented by sodium nitroprusside (100 microM), which releases NO spontaneously. This augmentation was prevented by hemoglobin (2 micrograms/ml), which binds the NO released by nitroprusside. Our previous experiments showed that norepinephrine-induced release of prostaglandin E2 is mediated by NO. Nitric oxidergic neurons were visualized in the median eminence adjacent to the LHRH terminals. The combined in vivo and in vitro results indicate that the pulsatile release of LHRH induced by norepinephrine is brought about by alpha 1-adrenergic activation of NO synthase. NO then induces prostaglandin E2 release that activates exocytosis of LHRH secretory granules into the portal vessels to induce pulsatile LH release.
Nitric oxide is a unique biological messenger molecule. It is produced by endothelial cells to mediate blood vessel relaxation; it mediates, in part, the immune functions of activated macrophages; and in the central and peripheral nervous system it serves as a neurotransmitter. In the nervous system, nitric oxide may regulate neurotransmitter release, it may play a key role in synaptic plasticity and morphogenesis, and it may regulate sexual and aggressive behavior. Under conditions of excessive formation, nitric oxide is emerging as an important neurotoxin.
This study was designed to examine the synthesis and function of nitric oxide during follicular development and luteinization in the rat ovary. Cells were obtained from hypophysectomized diethylstilbestrol-implanted immature female rats subjected to several hormonal regimens that resulted in preantral, Graafian, ovulatory, atretic, or luteinized ovaries. Cells obtained from ovaries at all stages of follicular development synthesized nitric oxide in a linear manner over time. The basal production of nitric oxide was 6- to 14-fold higher (P < 0.009) in cells obtained from luteinized ovaries than that in cells obtained from ovaries at all other developmental stages. Inhibiting endogenous nitric oxide synthesis in cells obtained from luteinized ovaries resulted in a 3-fold increase (P < 0.007) in the estradiol level without affecting progesterone synthesis. We used isoform-specific antisera to determine the cellular location and isoform(s) of nitric oxide synthase expressed in our cell culture system and in the luteinized ovary in vivo. Positive immunofluorescent staining for both the endothelial and inducible isoforms was observed in separate cell types. Immunoblotting experiments also showed that luteinized ovaries express the endothelial and inducible isoforms of nitric oxide synthase. Nitric oxide synthesis inhibits estradiol synthesis in vitro. We suggest that nitric oxide participates in functional luteal regression by inhibiting steroidogenesis.
Castration of adult male rats reduces by half the penile erectile response to electrical field stimulation (EFS) of the cavernosal nerve, and the activity of penile nitric oxide synthase (NOS). Both changes are prevented by androgen administration. We have now investigated whether other strategies of androgen ablation or competition may act as stronger inhibitors, and, if so, whether the stronger inhibition is due to the depletion of penile NOS content. Rats were castrated or left intact and were treated daily as follows: 1) intact, with the antiandrogen flutamide (25 mg/kg/day, i.p.); 2) castrated, with similar treatment; 3) castrated, with 17 beta-estradiol 3-benzoate (estradiol; via silastic tubing, s.c.). Additional groups of intact rats received injections of a GnRH antagonist (GnRHA, 1.25 mg/kg, s.c.), or were hypophysectomized and left untreated. Controls were untreated intact and castrated animals. After 7 days, rats were subjected to EFS, and the ratios between maximal intracavernosal pressure (MIP) and mean arterial pressure (MAP) were measured. Penile NOS activity and the contents of neuronal NOS (nNOS) and endothelial NOS (eNOS) were determined. Castration reduced the MIP:MAP ratio and penile NOS activity. Androgen receptor blockade with flutamide induced a similar response in intact rats. When flutamide treatment was combined with castration, the erectile response was nearly abolished, but NOS activity was not decreased below the values in castrated rats. Estradiol given to castrated rats and hypophysectomy or GnRHA treatment in intact rats diminished the erectile response below the level in castrated animals. In hypophysectomized rats, penile NOS activity fell below levels in castrated animals. contents of nNOS and eNOS were not significantly reduced by any treatment. These data suggest that penile erection in the rat is completely dependent on androgens, presumably because of their role in the maintenance of penile NOS activity and of other ancillary factors. However, only the complete blockade of residual androgen effects at the tissue level or a total androgen depletion can abolish the erectile response.
A review of the current literature is conducted to explore the developmental aspects, animal and human experiences and the effects of pharmacological manipulation to explain the role androgens play in sexual function with special emphasis on erectile function and the erectile tissue. This review reveals that androgens are necessary for the normal development of the penis and their deficiency results in significant structural abnormalities. Although androgen receptors in the penis decrease after puberty, they usually do not disappear completely. Animal data show that androgens support erectile function through a direct effect on the erectile tissue. Experimental castration results in impaired erectile response to central and peripheral stimulation and decrease in penile tissue concentration of nitric oxide synthase-containing nerves. Testosterone replacement reverses these abnormalities. In the rat penis, apoptosis is induced by castration and new DNA synthesis is induced by testosterone replenishment. Human data are less clear than animal data. Castration results in loss of libido and in erectile dysfunction. However, these effects are not universal. Testosterone enhances libido, frequency of sexual acts and sleep-related erections. Its effects on erotic erections are not clear.
Prior studies from this laboratory, using untreated castrated (CASTRATE) rats and testosterone-treated castrated (TESTO) rats, have shown that the magnitude of the intracavernosal pressure increase during erection is androgen dependent. Studies from this and other laboratories have also presented evidence suggesting that penile erection is mediated principally by nitric oxide (NO). The present report was designed to confirm that androgens maintain the availability of cavernosal NO and to determine if this androgenic action is exerted at the genomic level modulating the expression of the neuronal form of the nitric oxide synthase gene (nNOS). The results showed that administration of supplemental L-arginine failed to augment the erectile response in either group, suggesting that substrate availability is not a cause of the reduced response in CASTRATE animals. Inhibition of NO synthesis with a nitro-arginine competitive inhibitor of nitric oxide synthase enzyme protein (NOS) resulted in strong inhibition of erection in both TESTO and CASTRATE rats. When given in conjunction with ganglionic stimulation to induce erection, the NO releasing drug, sodium nitro-prusside (SNP), increased intracavernosal pressure in CASTRATE rats but not in TESTO rats, suggesting a deficiency of the available NO in CASTRATE-animals. Finally, reverse transcription-polymerase chain reaction (RT-PCR) demonstrated that mRNA levels for the enzyme nNOS in the penis were greater in TESTO animals than in CASTRATE rats. These results support the hypothesis that androgens mediate the erectile response in the rat penis by stimulating the expression of the neuronal isoform of nitric oxide synthase, thus maintaining an adequate supply of NO.
Expression and androgen regulation of the gene for neuronal nitric oxide synthase (NOS I) were examined in neurons of the major pelvic ganglia in male rats. Some of these postganglionic neurons innervate the penis and produce nitric oxide, which is believed to play a major role in penile erection. Rats were either castrated or sham operated and implanted with SILASTIC brand capsules filled with powdered testosterone (T) or 5alpha-dihydrotestosterone (5alphaDHT) or left empty. After 4 days, the number of neurons intensely stained for NADPH-diaphorase as well as those giving a NOS I signal in in situ hybridization experiments increased in castrated rats treated with testosterone by 31% and 42%, respectively, relative to those in untreated castrated rats. This suggests that the increase in NADPH-diaphorase activity resulted from enzyme synthesis and was due to a modification of NOS I messenger RNA (mRNA) accumulation. After 7 days, Northern blot analysis showed that castration produced a decrease in the amount of NOS I mRNA relative to that of ribosomal RNA. This decrease was almost prevented by T treatment. No significant differences were observed by reverse transcriptase-PCR between 7-day and 28-day treatments. However, in 7-day castrated rats treated with 5alphaDHT, NOS I signals relative to those of hypoxanthine phosphoribosyltransferase, taken as reference, were significantly higher than those in castrated rats and resembled those in sham-castrated rats, suggesting that 5alphaDHT was probably more potent than testosterone in preventing the decrease in NOS I mRNA levels elicited by castration. These results show that NOS I can be positively regulated by androgens and are consistent with the suggestion that these steroids play a role in the physiological processes of penile erection.
We investigated the effects of long-term testosterone replacement on copulatory behavior and dopaminergic neurotransmission in the medial preoptic area of aged male rats.
The rats were divided into 3 groups depending on testosterone replacement. Those in the long-term replacement group were castrated at the age of 12 months and received testosterone replacement thereafter for 12 months. In the short-term replacement group, rats were castrated at the age of 22 months and high or low dose testosterone replacement was done for 2 months. The control group consisted of aged rats 24 months old and young rats 12 weeks old, neither of which had been castrated or received testosterone replacement. We observed sexual behavior in rats of these groups. After a behavioral test, we measured the tissue concentration of dopamine in the MPOA and the change rate of the extracellular dopamine level induced by infusion of N-methyl-D-aspartic acid (NMDA) in the MPOA and compared the long-term replacement and no-replacement groups.
The rats in the long-term replacement group showed a mount rate at the same level as that of young rats at 6 weeks after starting replacement and it was maintained to 24 months of age. Their mount rate was significantly higher than that of the rats with the short-term replacement. A significantly higher change rate of dopamine release was recognized in the long-term group; however, no significant difference in the concentration of dopamine was recognized between aged rats with long-term replacement and those without replacement.
Aged rats (24 months old) with long-term testosterone replacement maintained almost the same level of mount behavior as young rats (12 weeks old). The results imply that long-term testosterone replacement may favorably alter the decline in the process of sexual activity with aging. The restoration by testosterone replacement of dopaminergic activity in the MPOA may be involved in the maintenance of sexual function in aged rats.
The purpose of this study was to investigate the in vivo effects of intracavernosal injections of adrenomedullin (ADM), calcitonin gene-related peptide (CGRP), nociceptin, vasoactive intestinal polypeptide (VIP), sodium nitroprusside (SNP), and prostaglandin E1 (PGE1) on penile erection in castrated and intact (control) anesthetized cats. Erectile responses to ADM, CGRP, nociceptin, VIP, SNP, and PGE1 were compared with responses to a standard triple-drug combination (1.65 mg of papaverine, 25 microg of phentolamine, and 0.5 microg of PGE1) in both castrated and control cats. In control animals, ADM, CGRP, nociceptin, VIP, SNP, and PGE1 induced penile erections similar to those elicited by the triple-drug combination. However, in castrated animals, there was a significant decrease in erectile response; the response to intracavernosal injection of the standard triple-drug combination in castrated cats was 28% of that of the control group of animals. Serum testosterone levels demonstrated a significant (P < 0.0001) positive correlation (r = 0.52) with intracavernosal pressure in response to the standard combination. A marked reduction in serum testosterone levels was observed in castrated cats when measured by radioimmunoassay (0.34 +/- 0.1 ng/dl in castrated cats, compared with 31.15 +/- 6 ng/dl in control cats). These data suggest that the presence of testosterone is a necessary prerequisite to sustain a pharmacologically induced penile erection in the cat.
This study was conducted to investigate the role of testosterone (T) in regulating the relaxation of isolated rat corpus cavernosum strips in vitro.
Male rats were divided into treatment groups of intact, castrate, and castrate with T replacement. Norepinephrine (NE) was added to contract each of the tissue strips. Next, sodium nitroprusside (SNP), experiment I, or 8-Br-cGMP, experiment II, was added to relax the cavernosum tissue. Percent relaxations were recorded for each treatment group at each dose level.
SNP was added to the NE-contracted tissues in doses of 10(-4) and 10(-3) M. In this experiment, castration significantly reduced tissue responsiveness to SNP and T replacement restored the response to intact levels. In the second experiment 8-Br-cGMP was added to the NE-contracted tissues in doses of 10(-5) and 10(-4) M. 8-Br-cGMP 10(-4) M was significantly less effective in relaxing tissue from castrate animals as compared with intact controls. Again, T treatment restored the response to intact levels.
Our results show a clear role of T in regulating the ability of corpus cavernosum tissue to relax when treated with SNP or 8-Br-cGMP in vitro. In addition, the data suggests that T regulates sites distal to the formation of cGMP in the relaxation pathway.
Ongoing studies in this laboratory have used the castrated rat, with and without testosterone replacement, to investigate how androgens maintain the erectile response. The high intracavernosal pressures during erection depend on both an increase in the rate at which blood flows into the sinuses of the corpus cavernosum and a decrease in the rate at which blood flows out (veno-occlusion). Accordingly, our studies investigated androgenic regulation of the arterioles that regulate inflow and of the intracavernosal muscle that regulates the veno-occlusive mechanism controlling outflow. The results of these studies show that castration causes a decline in the rate of inflow and that androgen replacement reverses this decline. The decline in inflow in the castrated rats is also reversed by the administration of a nitric oxide donor drug, suggesting that the androgen may regulate inflow by increasing the synthesis of nitric oxide. Testosterone also appears to regulate outflow by controlling the sensitivity of the erectile mechanisms to norepinephrine, considered to be the principle vaso-constrictor neurotransmitter in the erectile response. Taken together, the results of these studies suggest that androgens control the erectile response by altering the synthesis and action of the neurotransmitters that normally alter the state of contraction and relaxation of smooth muscle in the erectile tissue.
To determine the effectiveness of BAY 41-2272 on penile erections in an in vivo rabbit model. The nitric oxide (NO)-dependent increase of intracellular cyclic guanosine monophosphate (cGMP) by cGMP-phosphodiesterase (PDE5) inhibition has been shown to be an effective mechanism in the treatment of erectile dysfunction. Direct, NO-independent stimulation of soluble guanylyl cyclase should also lead to elevated cGMP levels in tissues and could be an attractive alternative therapeutic option for the treatment of erectile dysfunction. BAY 41-2272 is a novel non-NO-based direct stimulator of soluble guanylyl cyclase that activates purified enzyme in a synergistic fashion with NO.
BAY 41-2272 was administered to conscious rabbits intravenously (IV) and orally (PO). Erection was assessed in a time-dependent manner by measuring the length of the uncovered penile mucosa. Erections were evaluated in the absence and presence of NO (with intravenous sodium nitroprusside [SNP] as the NO donor).
BAY 41-2272 only induced weak penile erections in conscious rabbits after IV (1 mg/kg) and PO (10 mg/kg) administration in the absence of an NO donor. However, the efficacy of BAY 41-2272 was potentiated by the simultaneous administration of SNP. Through simultaneous SNP administration, the effective doses of BAY 41-2272 were reduced significantly (minimal effective dose 0.1 mg/kg IV and 1 mg/kg PO).
The results of this study clearly demonstrated the effect of BAY 41-2272 on penile erection in the conscious rabbit model after PO and IV administration. The time-course and onset of erection was concurrent with the stimulation by exogenous NO (SNP), suggesting that this new pharmacologic mechanism of soluble guanylyl cyclase stimulation could be used in the treatment of erectile dysfunction. Because the effect is increased by SNP, it can be expected that BAY 41-2272 would have enhanced activity during sexual arousal, when NO is produced endogenously.
Nitric oxide (NO), the main mediator of penile erection, is assumed to be synthesized in the penis by the neuronal constitutive nitric oxide synthase (nNOS). However, nNOS has not been identified in the penile smooth muscle, the target of NO action. The other NOS isozymes, the inducible NOS (iNOS) and the endothelial NOS (eNOS) have not been reported in any penile tissue. The smooth muscle vascular and trabecular tissue from rat corpora cavernosa is represented in vitro by cell cultures designated RPSMC. To determine whether iNOS can be expressed in penile smooth muscle, RPSMC were treated with different lymphokines and/or bacterial lipopolysaccharide (LPS). The selected inducer, LPS/interferon, elicited at 48 hours up to a 50-fold increase in nitrites in the medium; the nitroarginine methyl ester (L-NAME), aminoguanidine, actinomycin D, cycloheximide, transforming growth factor-beta1 (TGF-beta1), and dexamethasone, but was resistant to nifedipine and platelet-derived growth factor AB (PDGF-AB). iNOS induction increased with cell passage. The [3H]L-arginine/citrulline measurement of NO synthesis with intact cells confirmed these results. Incubations of soluble and particulate fractions showed that the cytosol contained most of the activity (Km = 43 microM), which was partially inhibited by ethyleneglycal-bis-tetraacetic acid (EGTA). The 4.4-kb iNOS mRNA peaked at a late period (24-30 hours) and remained high for up to 72 hours. iNOS mRNA induction was strongly inhibited by actinomycin D and dexamethasone, partially inhibited by TGF-beta1, inhibited slightly by PDGF-AB, and unaffected by nifedipine. These results show that iNOS can be expressed in RPSMC in a cell passage-dependent fashion that has so far not been reported for other cell lines, and that the induction reaches much higher levels than in rat or human vascular smooth muscle cells. The expression pattern is also distinctive for the penile cells in time course of induction, Ca2+ dependence, response to certain agents, and mRNA stability.
Edible bird's nest (EBN) is the nest of the swift that is made from its saliva. Although EBN has been widely used for enhancing immunocompetence, its antiviral efficacy has not been studied in detail. We found that EBN extract could strongly inhibit infection with influenza viruses in a host range-independent manner when it was hydrolyzed with Pancreatin F. Western blotting assay showed that the EBN extract bound to influenza virus. Furthermore, EBN extract could neutralize the infection of MDCK cells with influenza viruses and inhibit hemagglutination of influenza viruses to erythrocytes, but it could not inhibit the activity of influenza virus sialidase. Fluorometric HPLC indicated that the major molecular species of sialic acid in EBN is N-acetylneuraminic acid. The results suggest that EBN is a safe and valid natural source for the prevention of influenza viruses.
Erectile dysfunction affects over half of all men between 50 and 70 years of age, and by the age of 40, about 40% of men may suffer from some form of erectile dysfunction. Many disease states, such as diabetes, hypertension, depression, and vascular disease, are associated with the condition, which may occur many years prior to the onset of these disorders. The phenomenal success of sildenafil in improving erections in men with erectile dysfunction is due to the fact that the drug, as a phosphodiesterase inhibitor, improves the relaxation of smooth muscle cells, which become dysfunctional with the aging process. However, not everyone responds to this medication, mainly because the efficacy of the drug is directly dependent on the release of nitric oxide from the nerve terminals of the cavernosal nerve, and this may become defective with aging/certain disease states. The goal of gene therapy for organic impotence is to allow the patient to sustain physiologically elicited erections without resorting to pharmacological treatment immediately prior to the sexual act. Experimental efforts in gene therapy for erectile dysfunction are likely to continue intensively in a series of directions, some specific to the nature of the selected gene to be manipulated or the physiology of the corpora cavernosa itself, and others extrapolatable from the advancement of gene therapy in general.
The prevalence of erectile dysfunction increases as men age. Simultaneously, age related changes occur in male endocrine functioning. We examined the association between erectile dysfunction and total testosterone, bioavailable testosterone, sex hormone-binding globulin and luteinizing hormone.
Data were obtained from the Massachusetts Male Aging Study, a population based cohort study of 1,709 men. Self-reported erectile dysfunction was dichotomized as moderate or severe vs none or mild. Odds ratios and 95% CI were used to assess the association between sex hormone levels and erectile dysfunction. Multiple logistic regression models were used to adjust for potential confounders including age, body mass index, partner availability, phosphodiesterase type 5 inhibitor use, depression, diabetes and heart disease.
Using data from the most recent followup, analyses were conducted on 625 men with complete data. A moderate decrease in erectile dysfunction risk was observed with increasing total testosterone and bioavailable testosterone levels. However, this effect was not apparent after controlling for potential confounders. Increased luteinizing hormone levels (8 IU/l or greater) were associated with a higher risk of erectile dysfunction (adjusted OR 2.91, 95% CI 1.55-5.48) compared to luteinizing hormone levels less than 6 IU/l. A significant interaction between luteinizing hormone and total testosterone levels showed that increased testosterone levels were associated with a decrease in risk of erectile dysfunction among men with luteinizing hormone levels greater than 6 IU/l.
In this large population based cohort of older men we found no association among total testosterone, bioavailable testosterone, sex hormone-binding globulin and erectile dysfunction. Testosterone levels were associated with a decrease in risk of erectile dysfunction only in men with increased luteinizing hormone levels.
It is well known that testosterone enhances sexual interest leading to an increased frequency of sexual acts and an increase in the frequency of sleep-related erections. However, it has little effect on fantasy- or visually induced erections. Exact contribution to erection from testosterone in men remains unclear. Animal studies have well demonstrated that testosterone plays critical physiological (activity of nitric oxide synthases and phosphodiesterases), biochemical (through an endothelial-independent pathway and adrenergic tonicity) and structural (change of fibroelasticity and hollow cell accumulation) roles in erectile function. The supplementation of testosterone to castrated animals can restore erectile function. Clinically, reports of patients with erectile dysfunction (ED) combined with hypogonadism who receive testosterone therapy have inconsistent results. However, testosterone may ameliorate the expression of the phosphodiesterase-5 (PDE5) inhibitor, and the use of testosterone in conjunction with the PDE5 inhibitor revealed convincing results. Because of potential risks in clinical use, testosterone therapy should be individualized, carefully considered and closely monitored, especially, in patients with possible occult prostate cancer, and large benign prostatic hyperplasia. Lower urinary tract symptoms might be worsened by this treatment, since the prostate is an androgen-dependent tissue.
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A guide to birds' nest caves and birds' nests of Sarawak
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