ABSTRACT
Objective To investigate the biological effects of the Mucuna pruriens (M. pruriens) seed extracts that lacked L-DOPA, which was formerly reported as the active ingredient, on erectile dysfunction (ED) both in vitro and in vivo. Methods Seed of M. pruriens plant that cultivated in Mae Taeng District, Chiang Mai Province, Thailand, was collected. Component of its seeds were extracted and isolated into 2 fractions using methanol, polar and nonpolar. Each fraction was investigated for phytochemicals using gas chromatography and mass spectroscopy and was screened for biological activity in vitro using three different cell lines. The most biological active fraction was used to treat both streptozotocin (STZ)-induced diabetes mellitus-erectile dysfunction (DM-ED) male Wistar rats and normal rats (n = 6 per groups) to compare the effect on sexual behavior parameters, including number of intromission, mounting and ejaculation, with that of rats given Sildenafil by individually pairing with their female counterparts. Penile tissues and serums were collected to determine histological structure, related gene expression and biomolecules. Results The phytochemicals of the polar fraction were possibly catechol and its derivatives plus polyphenols, whereas the nonpolar fraction consisted of lipid derivatives. L-DOPA was not detected in either of the extracts. The polar fraction was able to up-regulate the expression of ED-related genes including eNOS and nNOS in vitro which subsequently promotes nitric oxide production and maintains intracellular cyclic guanosine monophosphate levels. When administrated to DM-ED rats, the polar extract significantly improved all sexual behavior parameters in DM-ED rats compared to untreated group (18.3 ± 1.8 to 10.8 ± 2.9 for intromission, 9.8 ± 2.2 to 5.7 ± 1.3 for mounting, and 1.8 ± 0.6 to 0.2 ± 0.4 for ejaculation). That effect might due to the ability of the extract to stimulate the expression of eNOS and nNOS which results in nitric oxide production and subsequently maintains cyclic guanosine monophosphate levels in penile tissue. Moreover, this extract may also prevent penile tissue deterioration due to diabetes. Conclusions The polar extract of M. pruriens seed can be used for ED therapy, especially in patients with metabolic diseases including diabetes. The action of the extract might be due to catechol and its derivatives and polyphenols.
ABSTRACT
OBJECTIVE@#To investigate the biological effects of the Mucuna pruriens (M. pruriens) seed extracts that lacked l-DOPA, which was formerly reported as the active ingredient, on erectile dysfunction (ED) both in vitro and in vivo.@*METHODS@#Seed of M. pruriens plant that cultivated in Mae Taeng District, Chiang Mai Province, Thailand, was collected. Component of its seeds were extracted and isolated into 2 fractions using methanol, polar and nonpolar. Each fraction was investigated for phytochemicals using gas chromatography and mass spectroscopy and was screened for biological activity in vitro using three different cell lines. The most biological active fraction was used to treat both streptozotocin (STZ)-induced diabetes mellitus-erectile dysfunction (DM-ED) male Wistar rats and normal rats (n = 6 per groups) to compare the effect on sexual behavior parameters, including number of intromission, mounting and ejaculation, with that of rats given Sildenafil by individually pairing with their female counterparts. Penile tissues and serums were collected to determine histological structure, related gene expression and biomolecules.@*RESULTS@#The phytochemicals of the polar fraction were possibly catechol and its derivatives plus polyphenols, whereas the nonpolar fraction consisted of lipid derivatives. l-DOPA was not detected in either of the extracts. The polar fraction was able to up-regulate the expression of ED-related genes including eNOS and nNOS in vitro which subsequently promotes nitric oxide production and maintains intracellular cyclic guanosine monophosphate levels. When administrated to DM-ED rats, the polar extract significantly improved all sexual behavior parameters in DM-ED rats compared to untreated group (18.3 ± 1.8 to 10.8 ± 2.9 for intromission, 9.8 ± 2.2 to 5.7 ± 1.3 for mounting, and 1.8 ± 0.6 to 0.2 ± 0.4 for ejaculation). That effect might due to the ability of the extract to stimulate the expression of eNOS and nNOS which results in nitric oxide production and subsequently maintains cyclic guanosine monophosphate levels in penile tissue. Moreover, this extract may also prevent penile tissue deterioration due to diabetes.@*CONCLUSIONS@#The polar extract of M. pruriens seed can be used for ED therapy, especially in patients with metabolic diseases including diabetes. The action of the extract might be due to catechol and its derivatives and polyphenols.
ABSTRACT
A monoclonal antibody to polysulphated polysaccharides was prepared and used in a plasma assay for heparin. The antibody reacted with heparin and semisynthetic heparin-like substances including dextran sulphate, pentosan polysulphate, and glycosaminoglycan polysulphate. The analytical method used a biotin-labeled monoclonal antibody in a competitive enzyme linked immunosorbent assay (ELISA). The level of heparin and heparin-like substances obtained by the assay correlated with those of a method based on the inhibition of Factor Xa, but were numerically different (y = -0.1747 + 1.0848x; p2 = 0.844). The assay could detect heparin and heparin-like substances in solution at a submicrogram or milliunit level, which suggested that it was suitable for pharmacokinetic studies of hepain and polysulfaled polysaccharides. The heparin concentrations in plasma from thalassemic patients, either with or without epitaxis, were not statistically different from those of normal subjects.
ABSTRACT
This study aimed to monitor chondroitin sulphate (CS; WF6 epitope) levels in crevicular fluid around maxillary molars and miniscrew implants during orthodontic molar intrusion. One miniscrew implant was placed in the midpalatal area of each of ten patients with open skeletal configurations, who required orthodontic molar intrusion, and two Sentalloy® closed-coil springs (100 g) were used for molar intrusion. Gingival crevicular fluid (GCF) around experimental and control molars, and peri-miniscrew implant crevicular fluid (PMICF) were collected before and during load application. Competitive ELISA with monoclonal antibody WF6 and colorimetric protein assay were used to detect CS (WF6 epitope), and total protein concentration, respectively.The results showed that the median CS (WF6 epitope) levels around experimental molars during the loaded period (12 weeks) (2.099 ng/µg of total protein) and those during each two-week interval of the loaded period (1.952, 1.854, 2.604, 2.414, 1.844, 1.44 ng/ µg of total protein respectively) were significantly greater than those during the unloaded period (2 weeks) (0.832 ng/ µg of total protein) (P
ABSTRACT
Abstract A periodate-resorcinol microassay for the quantitation of total sialic acid (TSA) has been developed. A 96-well microtiter plate format was used as a single reagent mixing well. The principle of this newly-developed assay is the periodate-resorcinol procedure. Conditions for the quantitation of sialic acid in serum, such as the concentration of periodate, resorcinol, incubation time and human serum volume, were optimized. It was found that the optimal concentration of periodic acid and resorcinol reagent were 1.3 mM and 0.6%, respectively. An incubation time of 60 minutes for the reaction of periodic acid and resorcinol with samples was found to give the highest absorbance. These new procedures were used for the quantitation of sialic acid in human serum. It was found that only 5 μL samples were needed to give a varied coefficient of the intraand inter-assay of 0.79% and 4.68%, respectively. Furthermore, this method demonstrated the recovery percentage of 94.25% by the addition of a known amount of pure sialic acid. This has been used to quantitate TSA in normal healthy and cancer serum and a significant difference between these two groups (p
ABSTRACT
Zingiber cassumunar Roxb. was previously shown to possess potent anti-inflammatory activity. We investigated the effect of its ethanol extract on hyaluronan (HA), an extracellular matrix compound with proinflammatory activity synthesis in human oral fibroblasts. Cultured fi broblasts were treated at various times with different concentrations of the extract (0.1-50 μg/mL) with or without retinoic acid (RA; 10 μM), or left untreated as a control. Culture medium was analyzed for HA quantities by the ELISA-based assay. Total RNA was harvested and RT-PCR analyses were performed to determine mRNA expression of hyaluronan synthase (HAS) -1, -2, and –3. Z. cassumunar extract, at 25 and 50 μg/mL, with or without 10 μM of RA, significantly decreased HA levels (p \< 0.05). Consistent with decreased HA levels, mRNA expression of HAS-2, but not HAS-1 or HAS-3, was selectively down-regulated by the extract. Collectively, these data indicate that the ethanol extract of Z. cassumunar inhibits HA synthesis in human oral fibroblasts, which may be involved in chronic inflammatory disorders, particularly in the oral cavity.
ABSTRACT
Twenty-five dogs were included in a randomized, double-blind trial to assess the efficacy of doxycycline (DOX) orally administered twice a day at 4 mg/kg/day (n = 12) for the treatment of osteoarthritis of the hip. Chondroitin sulfate (CS; 525 mg/day) was used as a positive control (n = 13). Dogs were re-examined monthly for 6 months after initiation of treatment. The assessment protocol included clinical score, radiographic findings and serum osteoarthritis biomarkers. Dogs treated with DOX showed statistically significant improvements (p < 0.05) in lameness, joint mobility, pain on palpation, weight-bearing and overall score at 2, 6, 4, 4 and 4 months, respectively, after treatment. Biomarker levels of CS-WF6 epitope and hyaluronan were significantly increased and decreased (p < 0.05) at 2 and 3 months after treatment compared to pretreatment. These results showed that DOX had a positive therapeutic effect in dogs with osteoarthritis.
Subject(s)
Animals , Dogs , Female , Male , Biomarkers/blood , Dog Diseases/drug therapy , Doxycycline/therapeutic use , Hyaluronic Acid/blood , Joints/drug effects , Lameness, Animal/drug therapy , Osteoarthritis, Hip/drug therapy , Time Factors , Treatment OutcomeABSTRACT
Hip dysplasia (HD) is one of the most important bone and joint diseases in dogs. Making the radiographic diagnosis is sometime possible when the disease has markedly progressed. Chondroitin sulfate (CS) and hyaluronan (HA) are the most important cartilage biomolecules that are elevated in the serum taken from dogs with osteoarthritis. The serum CS and HA can be detected by an ELISA technique, with using monoclonal antibodies against CS epitope 3B3 and WF6 and the HA chain as the primary antibodies. The aim of this study was to compare the levels of serum CS (both epitopes) and HA in non-HD and HD dogs. All 123 dogs were categorized into 2 groups. The non-HD group was composed of 98 healthy dogs, while the HD group was comprised of 25 HD dogs. Blood samples were collected for analyzing the serum CS and HA levels with using the ELISA technique. The results showed that the average serum level of the CS epitope WF6 in the HD group (2,594 +/- 3,036.10 ng/ml) was significantly higher than that in the non-HD group (465 +/- 208.97 ng/ml) (p < 0.01) while the epitope 3B3 in the HD group (105 +/- 100.05 ng/ml) was significantly lower than that in the non-HD group (136 +/- 142.03 ng/ml) (p < 0.05). The amount of serum HA in the HD group (134.74 +/- 59.71 ng/ml) was lower than that in the non HD group (245.45 +/- 97.84 ng/ml) (p < 0.05). The results indicate that the serum CS and HA levels might be used as biomarkers for osteoarthritis in HD dogs.