[show abstract][hide abstract] ABSTRACT: Traditional Indian systems of medicine use roots of Withania somnifera (W. somnifera) for impotence, infertility treatment, stress, and the aging process. Although W. somnifera improves semen quality by regulating reproductive hormone levels and oxidative stress, the molecular mechanism is not clear.
Our study uses high-resolution Nuclear Magnetic Resonance (NMR) spectroscopy to explore the scientific basis to reveal the pre- and post-treatment efficacy of W. somnifera on seminal plasma of infertile men-which remains unexplored to date.
A total of 180 infertile male patients were administered W. somnifera root powder at the rate of 5g/d for a three-month period. The study included age-matched, healthy men as a control (n=50) group. Proton NMR spectroscopy was used to measure lactate, alanine, glutamate, glutamine, citrate, lysine, choline, glycerophosphocholine (GPC), glycine, tyrosine, histidine, phenylalanine, and uridine in all seminal plasma samples. To appraise infertility levels, we also measured sperm concentration, motility, lipid peroxide, and hormonal perturbation.
W. somnifera therapy repairs the disturbed concentration of lactate, alanine, citrate, GPC, histidine, and phenylalanine in seminal plasma and recovers the quality of semen of post-treated compared to pre-treated infertile men. Serum biochemistry was also improved over post therapy in infertile men. Our findings reveal that W. somnifera not only reboots enzymatic activity of metabolic pathways and energy metabolism but also invigorates the harmonic balance of seminal plasma metabolites and reproductive hormones in infertile men.
The results suggest that W. somnifera may be used as an empirical therapy for clinical management and treatment of infertility.
Journal of ethnopharmacology 06/2013; · 2.32 Impact Factor
[show abstract][hide abstract] ABSTRACT: Urinary tract infection (UTI) is a potentially life-threatening infectious disease. For rapid directed therapy of UTIs, it is essential to determine the causative microorganism. To date, there is no single test that has been proven to reliably, rapidly and accurately identify the etiologic organism in UTI. The molecular methods for diagnosing the cause of UTI and prognostic development of clinically important metabolomic evaluations and their limitations for use in the diagnosis and monitoring of infections are discussed in this review article. The application of the emerging investigative device NMR spectroscopy as a surrogate method for the diagnosis of UTI is also addressed.
[show abstract][hide abstract] ABSTRACT: To address the shortcomings of urine culture for the rapid identification of urinary tract infection (UTI), we applied (1)H-nuclear magnetic resonance (NMR) spectroscopy as a surrogate method for fast screening of microorganisms. Study includes 682 urine samples from suspected UTI patients, 50 healthy volunteers, and commercially available standard strains of gram negative bacilli (GNB) (Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Enterobacter, Acinetobacter, Proteus mirabilis, Citrobacter frundii) and gram positive cocci (GPC) (Enterococcus faecalis, Streptococcus group B, Staphylococcus saprophyticus). Acetate, lactate, ethanol, succinate, creatinine, trimethylamine (TMA), citrate, trimethylamin-N-oxide, glycine, urea, and hippurate were measured by (1)H NMR spectroscopy. All urine specimens were evaluated with culture method. Multivariate discriminant function analysis (DFA) reveals that acetate, lactate, succinate, and formate were able to differentiate, with high accuracy (99.5%), healthy controls from UTI patients. This statistical analysis was also able to classify GNB to GPC infected urine samples with high accuracy (96%). This technique appears to be a promising, rapid, and noninvasive approach to probing GNB and GPC infected urine specimens with its distinguishing metabolic profile. The determination of infection will be very important for rapidly and efficiently measuring the efficacy of a tailored treatment, leading to prompt and appropriate care of UTI patients.
Journal of Proteome Research 03/2012; 11(3):1844-54. · 5.06 Impact Factor
[show abstract][hide abstract] ABSTRACT: Proanthocyanidin is commonly used for inhibiting urinary tract infection (UTI) of sensitive strains of Escherichia coli. The aim of this study was to investigate the effect of proanthocyanidin on adherence of uropathogenic multi-drug resistant E. coli to uroepithelial cells, which has not yet been investigated so far. Extracts of the purified proanthocyanidin were prepared from dried cranberry juice. Purity and structural assignment of proanthocyanidin was assessed using high performance liquid chromatography and (13)C nuclear magnetic resonance spectroscopy, respectively. Subsequently, its affect on multi-drug resistant bacteria as well as quantification of anti-adherence bioactivity on human vaginal and bladder epithelial cells was appraised. Inhibition of adherence to an extent of about 70% with multi-drug resistant E. coli strains was observed on uroepithelial cell. The anti-adherence bioactivity of the proanthocyanidin was detected at concentrations of 10-50 µg/ml with significant bacteriuria. Probable proanthocyanidin through A-type linkages either combines to P-fimbriae of bacterial cells or modifies the structural entity of P-fimbriae and inhibits bacterial adherence to uroepithelial cells. The proanthocyanidin exhibited anti-adherence property with multi-drug resistant strains of uropathogenic P-fimbriated E. coli with in vitro study. Hence proanthocyanidin may be considered as an inhibitory agent for multi-drug resistant strains of E. coli adherence to uroepithelial cells.
Urological Research 06/2011; 40(2):143-50. · 1.59 Impact Factor
[show abstract][hide abstract] ABSTRACT: The objective of this study was to employ proton nuclear magnetic resonance ((1)H NMR) spectroscopy to evaluate the impact of Mucuna pruriens seeds on the metabolic profile of seminal plasma of infertile patients. A total of 180 infertile patients were administered M. pruriens seed powder for a period of three months. Age-matched healthy men comprised the control (n=50) group in the study. Lactate, alanine, choline, citrate, glycerophosphocholine (GPC), glutamine, tyrosine, histidine, phenylalanine, and uridine were measured in seminal plasma by (1)H NMR spectroscopy. To evaluate the degree of infertility and extent of hormonal imbalance induced by this milieu, separate sperm concentration, motility, lipid peroxide in seminal plasma and LH, FSH, T, and PRL hormone concentration in serum were measured using standard laboratory methods and RIA, respectively, in the same subjects. M. pruriens therapy rectifies the perturbed alanine, citrate, GPC, histidine and phenylalanine content in seminal plasma and improves the semen quality of post-treated infertile men with compared to pre-treated. Concomitantly, clinical variables in seminal plasma and blood serum were also improved over post therapy in infertile men. On the basis of these observations, it may be proposed that M. pruriens seed powder not only reactivates the enzymatic activity of metabolic pathways and energy metabolism but also rejuvenates the harmonic balance of male reproductive hormones in infertile men. These findings open more opportunities for infertility treatment and management by improving semen quality.
Journal of pharmaceutical and biomedical analysis 04/2011; 55(5):1060-6. · 2.45 Impact Factor
[show abstract][hide abstract] ABSTRACT: Traditional seminal fluid-based clinical descriptors used to predict infertility and sub-fertility have shortcomings, including lack of insight into the underlying pathology. These methods are also time-consuming and labor-intensive. To address these limitations, (1)H nuclear magnetic resonance (NMR) spectroscopy was used to identify and classify signature biomarkers. Semen samples collected from 60 healthy, fertile men and from 125 infertile (normozoospermic and oligozoospermic) patients. Lactate, alanine, choline, citrate, glycerophosphocholine (GPC), glutamine, tyrosine, histidine, phenylalanine, and uridine were measured by (1)H NMR spectroscopy. The sperm concentration, motility, lipid peroxidation, and total protein were evaluated with standard laboratory methods in the same samples. NMR-quantified metabolites and clinical laboratory data were analyzed, separately, through linear multivariate discriminant function analysis (DFA) to determine the signature descriptors for each group. DFA reveals that alanine, citrate, GPC, tyrosine, and phenylalanine can be used to determine infertility. DFA-based classification demonstrated high accuracy (92.4% by NMR and 94.1% by clinical laboratory method) in differentiating healthy controls from infertile patients. This statistical analysis was also able to accurately classify normozoospermic to oligozoospermic samples (92.9% by NMR and 92.6% by clinical laboratory method). In conclusion, (1)H NMR-based metabolic screening appears to be a promising, rapid, and non-invasive approach to probing infertility that has similar sensitivity and specificity to the tedious laboratory method.
Journal of pharmaceutical and biomedical analysis 01/2011; 54(1):106-13. · 2.45 Impact Factor
[show abstract][hide abstract] ABSTRACT: To address the shortcomings of urine culture for the diagnosis of urinary tract infection (UTI), we used 1H-nuclear magnetic resonance (NMR) spectroscopy for identifying and quantifying Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia and Proteus mirabilis.
Urine samples from patients with suspected UTI (617), healthy volunteers (50) and commercially available standard strains of E. coli, K. pneumonia, P. aeruginosa, Enterobacter, Acinobacter, Pr. mirabilis, Citrobacter frundii, Streptococcus saprophyticus and Enterococcus faecalis were assessed between 2003 and 2006. 1H-NMR spectra were recorded on a 400 MHz spectrophotometer; to quantify the bacteria we estimated the areas under the spectral peaks of the specific metabolic product compared with the known concentration of trimethyl silyl propionic acid. All urine specimens were cultured in addition to an assessment by NMR spectroscopy.
Preliminary urinary spectroscopy of the unprocessed samples showed peaks of nonspecific metabolites such as succinate, acetate, lactate and ethanol, indicating infected samples. Based on the results from processed samples, 93% (240/256) of E. coli, 92% (101/110) of K. pneumoniae, 93% (56/60) of P. aeruginosa and eight of 10 Pr. mirabilis could be diagnosed with NMR (numerator) and urine culture (denominator). The remaining samples were sterile and/or had a bacterial population of <10(3) colony-forming units (CFU)/mL. The NMR method diagnosed bacterial densities of >10(3) CFU.
The identification of the common uropathogens E. coli, K. pneumoniae, P. aeruginosa and Pr. mirabilis by NMR spectroscopy has a shorter reporting time and can be used to differentiate between infected, contaminated and sterile specimens.
BJU International 03/2009; 104(2):236-44. · 3.05 Impact Factor
[show abstract][hide abstract] ABSTRACT: The toxic effects of Al(3+) have been studied in 90-days AlCl(3) orally treated male albino rats (n = 7) using (1)H NMR spectroscopy-based metabolic profile of rat serum and urine, serum enzyme tests, behavioral impairment, and histopathology of kidney and liver. Metabolic profile of 90-days Al(3+)-treated rat sera showed significantly elevated levels of alanine, glutamine, beta-hydroxy-butyrate, and acetoacetate and significantly decreased level of acetone when compared with that of control rats. However, metabolic profile of 90-days Al(3+)-treated rat urine showed significantly decreased levels of citrate, creatinine, allantoin, trans-aconitate, and succinate and significantly increased level of acetate when compared to control rats. The overall perturbations observed in the metabolic profile of serum and urine demonstrate the impairment in the tricarboxylic acid cycle, liver and kidney metabolism, which was further reinstated by clinical chemistry and histopathological observations. Moreover, "in vivo" behavioral impairment has also been observed as the indication of aluminum neurotoxicity.
Journal of Biochemical and Molecular Toxicology 02/2008; 22(2):119-27. · 1.60 Impact Factor
[show abstract][hide abstract] ABSTRACT: The (1)H NMR spectroscopic method is suggested and its utility is demonstrated for the diagnosis of Klebsiella pneumoniae (K. pneumoniae) in urinary tract infection (UTI). K. pneumoniae have the specific property of metabolizing glycerol to 1,3-propanediol (1,3-PD), acetate, ethanol and succinate. The quantity of 1,3-PD produced correlates well with the viable bacterial count. Other common bacteria causing UTI (except for Citrobacter frundii), such as Escherichia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa), Enterobacter aerogenes, Acinetobacter baumanii, Proteus mirabilis, Enterococcus faecalis, Streptococcus gp B and Staphylococcus aureus do not metabolize glycerol under similar conditions. Citrobacter frundii (C. frundii) also gives the same NMR results but is easily differentiated as being motile on direct microscopic examination of urine and it is not common nosocomial infectious agent in urinary tract infection. The method provides a single-step documentation of K. pneumoniae (and C. frundii) qualitatively as well as quantitatively. Out of the total 614 subjects considered, clinical diagnosis of UTI was obtained in 516 cases (84%). The NMR-based screening had a sensitivity of 90%, a specificity of 100% and a false negativity of 10% relative to the conventional quantitative culture method. In the present authors' experience, the results of NMR spectroscopy based screening show a very good correlation with the diagnosis of urinary tract infected patients.
NMR in Biomedicine 01/2007; 19(8):1055-61. · 3.45 Impact Factor
[show abstract][hide abstract] ABSTRACT: A high-resolution (1)H NMR study of serum and urine of fulminant hepatic failure patients (n = 22) [surviving (n = 12) and non-surviving (n = 10)] is reported. Glutamine in serum and urine glutamine:creatinine ratio were higher in non-surviving patients compared with surviving patients [serum glutamine, 3.08 (1.68-7.11) vs 0.56 (0.34-0.99) mM, median and range; p = 0.0001 and urine glutamine:creatinine ratio, 1.72 (0.24-7.76) vs 0.39 (0.1-0.84), p = 0.1], and urine urea:creatinine ratio was higher in surviving patients compared with non-surviving patients [10.83 (0.2-22.6) vs 2.09 (0.96-4.0), p = 0.002]. On the other hand, no significant differences were found in the conventionally employed clinical parameters such as serum alanylaminotransferase, aspartylaminotransferase and bilirubin except prothrombin time (p = 0.02). The difference in serum glutamine and urine urea was significant in the two categories of patients and distinctly different values of serum glutamine for both the categories of patients correctly predicted the outcome. These results promise immense potential for NMR spectroscopy in rapidly deciding on the need for advanced therapeutic intervention such as artificial liver support or emergency liver transplantation in FHF.
NMR in Biomedicine 09/2006; 19(5):521-6. · 3.45 Impact Factor
[show abstract][hide abstract] ABSTRACT: The utility of (1)H NMR spectroscopy is suggested and demonstrated for the diagnosis of Pseudomonas aeruginosa in urinary tract infection (UTI). The specific property of P. aeruginosa of metabolizing nicotinic acid to 6-hydroxynicotinic acid (6-OHNA) is exploited. The quantity of 6-OHNA produced correlates well with the viable bacterial count. Other common bacteria causing UTI such as Escherichia coli, Klebsiella pneumonia, Enterobacter aerogenes, Acinetobacter baumanii, Proteus mirabilis, Citrobacter frundii, Enterococcus faecalis, Streptococcus gp B and Staphylococcus aureus do not metabolize nicotinic acid under similar conditions. The method provides a single-step documentation of P. aeruginosa qualitatively as well as quantitatively. The NMR method is demonstrated on urine samples from 30 patients with UTI caused by P. aeruginosa.
NMR in Biomedicine 09/2005; 18(5):293-9. · 3.45 Impact Factor
[show abstract][hide abstract] ABSTRACT: This article describes proton MR spectroscopic analysis of cerebrospinal fluid of 167 children suffering from meningitis and 24 control cases. Quantification of 12 well-separated and commonly observed cerebrospinal fluid metabolites viz., beta-hydroxybutyrate, lactate, alanine, acetate, acetone, acetoacetate, pyruvate, glutamine, citrate, creatine/creatinine, glucose (total) and urea was carried out using Bruker's NMRQUANT software with respect to a known concentration of sodium-3-(trimethylsilyl)-2,2,3,3-d4-propionate (TSP), serving as an external reference. The assignment of urea in CSF is reported for the first time by NMR. The presence of cyclopropane, observed for the first time in tuberculous meningitis overall in 85.1% of cases, acts as a finger-print marker for the differential diagnosis. Multivariate discriminant function analysis was carried out for the proton MR-detected metabolite information and the clinical symptoms data of the meningitis and control cases to find the important descriptors for classification, followed by a re-validation of the entire database. It was found that the control could be differentiated from the disease group with a success rate of 96.4%, followed by the differential diagnosis of tuberculous meningitis with a corresponding value of 77.2%. Excluding the presence of cyclopropane, bacterial meningitis could be classified 84.4% correct and viral meningitis with a rate of 83.3%. It is proposed that the NMR spectroscopic information, along with other routine clinical features, may serve as an additional diagnostic tool for the differential diagnosis of meningitis in children.
NMR in Biomedicine 07/2005; 18(4):213-25. · 3.45 Impact Factor
[show abstract][hide abstract] ABSTRACT: High-resolution 1H-NMR spectroscopy of serum and urine samples of an 11-year-old male living related orthotopic liver transplant recipient is reported. Serum glutamine increased to abnormal levels along with simultaneous abnormal excretion of urinary glutamine post-transplantation. High levels of glutamine in both blood and urine and concomitant reduced urea levels in urine were found to be evidence of impairment in urea cycle and compatible with persistently abnormal graft function. Thus glutamine levels in serum and urine, and urea in the urine as observed by 1H-NMR spectroscopy highlight their important roles in monitoring liver graft function; increased glutamine levels lead to brain damage, if untreated.
NMR in Biomedicine 07/2003; 16(4):185-8. · 3.45 Impact Factor