Non-enzymatic glycoxidation linked with nutrition enhances the tumorigenic capacity of prostate cancer epithelia through AGE mediated activation of RAGE in cancer associated fibroblasts.

The molecular implications of food consumption on cancer etiology are poorly defined. The rate of nutrition associated non-enzymatic glycoxidation, a reaction that occurs between reactive carbonyl groups on linear sugars and nucleophilic amino, lysyl and arginyl groups on fats and proteins, is rapidly increased by food cooking and manufacturing processes. In this study, we assign nutrition-associated glycoxidation with significant oncogenic potential, promoting prostate tumor growth, progression, and metastasis in vivo. Advanced glycation end products (AGEs) are the final irreversible product of non-enzymatic glycoxidation. Exogenous treatment of prostate tumor cells with a single AGE peptide replicated glycoxidation induced tumor growth in vivo. Mechanistically, receptor for AGE (RAGE) deficiency in the stroma inhibited AGE mediated tumor growth. Functionally, AGE treatment induced RAGE dimerization in activated fibroblasts which sustained and increased the migratory potential of tumor epithelial cells. These data identify a novel nutrition associated pathway that can promote a tissue microenvironment conducive for aggressive tumor growth. Targeted and/or interventional strategies aimed at reducing AGE bioavailability as a consequence of nutrition may be viewed as novel chemoprevention initiatives.

Efficacy of Tea Tree Oil in the Treatment of Equine Streptothricosis.

Streptothricosis is a dermatitis characterized by matted tufts of hair and coalescing, pustular crusts that affects many livestock species, including horses. It results from cutaneous infection by the actinobacterium Dermatophilus congolensis. For economic reasons, the ailment is often treated with commercially available over-the-counter (OTC) products or home remedies rather than prescribed medications. This study aimed to determine the efficacy of tea tree oil (TTO), an essential oil of Melaleuca alternifolia, as an OTC treatment for streptothricosis. Bacteria were isolated from presumptive streptothricosis lesions on horses at a farm in Forest, Virginia. These isolates were microbiologically and genetically confirmed to be D. congolensis. The antimicrobial activity of TTO against D. congolensis isolates was determined by minimum inhibitory concentration and disc diffusion assays and compared with three OTC products advertised specifically for the treatment of "rain rot," a colloquial term for streptothricosis. A 1% TTO solution (v/v, in baby oil) and the three selected OTC products were applied to equine streptothricosis lesions to evaluate in vivo resolution of the lesions. Tea tree oil exhibited antimicrobial behavior against D. congolensis in vitro and produced marked improvement of streptothricosis lesions in vivo. These results have implications for development of TTO as a possible treatment for streptothricosis.

From 20th century metabolic wall charts to 21st century systems biology: database of mammalian metabolic enzymes.

The organization of the mammalian genome into gene subsets corresponding to specific functional classes has provided key tools for systems biology research. Here, we have created a web-accessible resource called the Mammalian Metabolic Enzyme Database (https://hpcwebapps.cit.nih.gov/ESBL/Database/MetabolicEnzymes/MetabolicEnzymeDatabase.html) keyed to the biochemical reactions represented on iconic metabolic pathway wall charts created in the previous century. Overall, we have mapped 1,647 genes to these pathways, representing ~7 percent of the protein-coding genome. To illustrate the use of the database, we apply it to the area of kidney physiology. In so doing, we have created an additional database (Database of Metabolic Enzymes in Kidney Tubule Segments: https://hpcwebapps.cit.nih.gov/ESBL/Database/MetabolicEnzymes/), mapping mRNA abundance measurements (mined from RNA-Seq studies) for all metabolic enzymes to each of 14 renal tubule segments. We carry out bioinformatics analysis of the enzyme expression pattern among renal tubule segments and mine various data sources to identify vasopressin-regulated metabolic enzymes in the renal collecting duct.

A knowledge base of vasopressin actions in the kidney.

Biological information is growing at a rapid pace, making it difficult for individual investigators to be familiar with all information that is relevant to their own research. Computers are beginning to be used to extract and curate biological information; however, the complexity of human language used in research papers continues to be a critical barrier to full automation of knowledge extraction. Here, we report a manually curated knowledge base of vasopressin actions in renal epithelial cells that is designed to be readable either by humans or by computer programs using natural language processing algorithms. The knowledge base consists of three related databases accessible at https://helixweb.nih.gov/ESBL/TinyUrls/Vaso_portal.html. One of the component databases reports vasopressin actions on individual proteins expressed in renal epithelia, including effects on phosphorylation, protein abundances, protein translocation from one subcellular compartment to another, protein-protein binding interactions, etc. The second database reports vasopressin actions on physiological measures in renal epithelia, and the third reports specific mRNA species whose abundances change in response to vasopressin. We illustrate the application of the knowledge base by using it to generate a protein kinase network that connects vasopressin binding in collecting duct cells to physiological effects to regulate the water channel protein aquaporin-2.

Evolutionary comparisons suggest many novel cAMP response protein binding sites in Escherichia coli.

The cAMP response protein (CRP) is a transcription factor known to regulate many genes in Escherichia coli. Computational studies of transcription factor binding to DNA are usually based on a simple matrix model of sequence-dependent binding energy. For CRP, this model predicts many binding sites that are not known to be functional. If they are indeed spurious, the underlying binding model is called into question. We use a species comparison method to assess the functionality of a population of such predicted CRP sites in E. coli. We compare them with orthologous sites in Salmonella typhimurium identified independently by CLUSTALW alignment, and find a dependence of mutation probability on position in the site. This dependence increases with predicted site binding energy. The positions where mutation is most strongly suppressed are those where mutation would have the biggest effect on predicted binding energy. This finding suggests that many of the novel sites are functional, that the matrix model correctly estimates their binding strength, and that calculated CRP binding strength is the quantity that is conserved between species. The analysis also identifies many new E. coli binding sites and genes likely to be functional for CRP.

The effects of sevoflurane on serum creatinine and blood urea nitrogen concentrations: a retrospective, twenty-two-center, comparative evaluation of renal function in adult surgical patients.

UNLABELLED: Despite mounting clinical evidence that supports its safety, the question of the potential adverse effects of sevoflurane on renal function continues to generate some controversy. This study retrospectively evaluated pooled renal laboratory data from 22 different clinical trials that compared sevoflurane with three widely used anesthetics. The trials examined postoperative changes in serum creatinine and blood urea nitrogen levels from a total of 3, 436 ASA physical status I-IV adult surgical patients administered either sevoflurane (n = 1941) or a control drug (isoflurane, enflurane, or propofol; n = 1495) as the maintenance anesthetic. The incidences of increased serum creatinine and blood urea nitrogen concentrations were similar among patients administered sevoflurane and those administered control drugs. Additionally, no trends specific to sevoflurane were observed with respect to postoperative serum creatinine concentration and fresh gas flow rate, concurrent treatment with nephrotoxic antibiotics, or type of carbon dioxide absorbent. IMPLICATIONS: Our data for changes in serum creatinine and blood urea nitrogen indicate that, for exposures of less than 4 minimum alveolar anesthetic concentration/h, sevoflurane is not associated with an increased risk of renal toxicity compared with other commonly used anesthetics. For clinical purposes, the pre- to postoperative changes in serum creatinine and blood urea nitrogen are appropriate measures of renal function in surgical patients.

Comparison of emergence and recovery characteristics of sevoflurane, desflurane, and halothane in pediatric ambulatory patients.

This study compares the emergence and recovery characteristics of sevoflurane, desflurane, and halothane in children undergoing adenoidectomy with bilateral myringotomy and the insertion of tubes. Eighty children 1-7 yr of age were studied. Thirty minutes prior to the induction of anesthesia, all patients received 0.5 mg/kg midazolam orally. Patients were randomly assigned to one of four groups: Group 1, sevoflurane induction and maintenance (S:S); Group 2, halothane induction and sevoflurane maintenance (H:S); Group 3, halothane induction and maintenance (H:H); or Group 4, halothane induction and desflurane maintenance (H:D). Tracheal intubation was facilitated with the use of a single dose of 0.2 mg/kg mivacurium. A Mapelson D circuit was used, and all patients received N2O:O2 60:40 for induction and maintenance at standardized appropriate fresh gas flow. Ventilation was controlled to maintain normocapnia. End-tidal concentration of anesthetics was maintained at approximately 1.3 minimum alveolar anesthetic concentration (MAC) (halothane: 0.56; sevoflurane: 2.6; desflurane: 8.3) until the end of surgery when all anesthetics were discontinued. Emergence (extubation), recovery (Steward score 6), and discharge times were compared among patients in the four groups using analysis of variance and Newman-Keuls tests P < 0.05 was considered significant. There were no significant differences among the four groups with respect to age, weight, duration of surgery, or duration of anesthesia. Emergence and recovery from anesthesia were significantly faster in the desflurane group (Group 4) compared with the sevoflurane and halothane groups (Groups 1, 2, and 3) (5 +/- 1.6 min vs 11 +/- 3.7, 11 +/- 4.0, 10 +/- 4.0 min and 11 +/- 3.9 min vs 17 +/- 5.5, 19 +/- 7.1, 21 +/- 8.5 min, respectively). There was a significantly greater incidence of postoperative agitation and excitement in patients who received desflurane (55%) versus sevoflurane (10%) and halothane (25%). There were no significant differences among the four groups with respect to the time to meet home discharge criteria (134 +/- 36.9, 129 +/- 53.3, 117 +/- 64.6, 137 +/- 22.6 in Groups 1, 2, 3, and 4, respectively), in the time to drink oral fluids (139 +/- 31.6, 136 +/- 53.8, 123 +/- 65.0, 142 +/- 29.4 min, respectively), or in the incidence of postoperative vomiting. It is concluded that, although desflurane resulted in the fastest early emergence from anesthesia, it was associated with a greater incidence of postoperative agitation. Sevoflurane resulted in similar emergence and recovery compared with halothane. Desflurane and sevoflurane did not result in faster discharge times than halothane in this patient population.

The predictive value of an initial serum beta human chorionic gonadotropin level for pregnancy outcome following in vitro fertilization.

PURPOSE: Our purpose was to assess the predictive value for pregnancy outcome of an initial serum quantitative beta-hCG measurement obtained 11 or 12 days after embryo transfer in an in vitro fertilization program. METHODS: A prospective, descriptive study of 153 pregnancies achieved via in vitro fertilization-embryo transfer was performed. Initial beta-hCG levels and subsequent pregnancy outcomes were compared. RESULTS: The overall mean initial beta-hCG level was 91 +/- 85.8 mIU/ml for normal (singleton, multiple-gestation) pregnancies and 29 +/- 24.9 mIU/ml for abnormal (miscarriage, ectopic) pregnancies (P < 0.01; power, > 80%). While 93.9% of patients with initial beta-hCG levels > or = 42 mIU/ml had normal pregnancies, 56.4% of those with initial levels < 42 mIU/ml experienced abnormal outcomes. With 42 mIU/ml as the cutoff level for predicting a normal pregnancy, this screening test yielded a sensitivity of 79.3% and a specificity of 83.8%. CONCLUSIONS: Initial serum beta-hCG levels obtained 11 or 12 days after embryo transfer may be predictive of pregnancy outcome in an in vitro fertilization program.

Inhalation toxicity study of a haloalkene degradant of sevoflurane, Compound A (PIFE), in Sprague-Dawley rats.

BACKGROUND: Under certain circumstances in the clinical setting, contact of the anesthetic sevoflurane with a CO2 absorbent (e.g., soda lime, Baralyme) leads to the formation of a degradant designated as pentafluoroisopropenyl fluoromethyl ether (PIFE; Compound A). Previous studies have shown that the kidney is the primary target organ for toxicity in the rat. This study was designed to determine the impact of PIFE on rat renal histology correlated with functional changes. The findings are discussed in terms of probable mechanism of action and relevance to humans. METHODS: Male and female Sprague-Dawley rats were exposed to 0, 30, 61, 114, or 202 ppm PIFE for a single 3-h period via nose-only inhalation. Rats were observed daily for behavioral changes or external physical signs of toxicity (i.e., lacrimation, dyspnea, piloerection, etc.) and body weights were recorded at 6, 4, and 1 day preexposure and 1, 3, 7, and 13 days postexposure. Animals were evaluated for hematologic, clinical chemistry and/or urinalysis changes immediately postexposure and/or at 1, 4, and 14 days postexposure. Rats were killed, subjected to a macroscopic postmortem examination, and evaluated for histopathologic changes in all major tissues and organs at 1, 4, and 14 days postexposure. RESULTS: Labored breathing was observed in 3 of the 20 and 2 of the 20 rats in the 114 ppm and 202 ppm groups, respectively, during the 3-h exposure period. No significant reductions in body weight gain were noted during the 2-week study period. Clinical chemistry evaluations revealed increases in blood urea nitrogen and creatinine 1 day postexposure in males and females exposed to 202 ppm PIFE. Changes in urinary glucose, protein and N-acetyl-beta-glucoaminidase/creatinine were evident one day postexposure in males and females exposed to 202 ppm and in males exposed to 114 ppm PIFE. Most values were within normal ranges by 4 or 14 days postexposure. No drug-related alterations in hematologic parameters were noted. Evidence of olfactory epithelial degeneration and desquamation in the nasal turbinates was noted at 4 days postexposure in male and female rats exposed to 202 ppm PIFE. Concentration-dependent renal tubular necrosis and tubular cell hyperplasia, in the corticomedullary border, were observed in males and females exposed to 114 and 202 ppm PIFE. The severity of tubular necrosis in both males and females was considered minimal to slight at the 114 ppm exposure concentration and slight to moderate at the 202 ppm exposure. Both the numbers of affected animals and severity were reduced over time. The most marked changes in serum and urine chemistry were associated with the animals described as having moderate renal necrosis. Male rats appeared more susceptible to nephropathy than female rats. There were no other PIFE-related histopathologic findings. CONCLUSIONS: The renal histopathologic findings in this study are consistent with those reported in previous acute studies in rats after PIFE administration. Functional changes in the kidney, as evidenced by serum chemistry and urinalyses, were observed at exposure concentrations that induced morphologic alterations.

A comparative prospective study using matched samples to determine the influence of subnormal hypo-osmotic test scores of spermatozoa on subsequent fertilization and pregnancy rates following in-vitro fertilization.

The achievement of pregnancies in vivo is rare in couples where the male partner has defective sperm membranes as shown by hypo-osmotic swelling (HOS) test scores of < 50%. However, there have been mixed reports on the value of the HOS test in predicting outcome following invitro fertilization; some studies suggest reduced fertilization rates and others find little, if any, predictability of decreased fertilization. The assumption has been made that fertilization rates are proportional to pregnancy rates; however, this may not necessarily be true since defective spermatozoa could lead to a less viable pre-embryo and therefore a decreased viable pregnancy rate. We performed a comparative prospective study using matched controls to evaluate fertilization rates and to determine subsequent pregnancy rates. The mean HOS scores were 70.0 and 36.7% respectively, with mean motile sperm concentrations of 35.7 and 34.0 x 10(6)/ml in 27 matched pairs. There was no difference in the mean number of oocytes retrieved, fertilization rates or number of embryos transferred between the two groups by HOS score. The clinical and viable pregnancy rates and implantation rates were 25.9, 18.5 and 9.9% for normal versus 3.7, 3.7 and 1.1% for subnormal groups. These data suggest that low HOS scores may be associated with the formation of defective embryos, leading to low pregnancy rates but normal fertilization rates.

The effect of sera antisperm antibodies in the female partner on in vivo and in vitro pregnancy and spontaneous abortion rates.

PROBLEM: To determine the incidence of antisperm antibodies (ASA) in female sera from infertile couples or those suffering from recurrent abortions. Also to determine if the pregnancy and/or abortion rates are any higher in those positive versus those negative for ASA. METHOD: All registered patients had sera drawn and ASA measured by indirect immunobead test on initial study. Pregnancy and abortion rates were determined for patients undergoing in vivo or in vitro therapy. RESULTS: There was a low incidence of ASA in patients having in vivo or in vitro treatment. There was no decrease in pregnancy rates (PRs) or increase in spontaneous abortions (SAB) in those positive for ASA. CONCLUSION: Antisperm antibodies in female sera do not seem to be etiologic in causing infertility or SAB. Future studies might consider changing the antigen source from donor sperm to husband's sperm.

Pregnancy rates in donors versus recipients according to the serum progesterone level at the time of human chorionic gonadotropin in a shared oocyte program.

Previous data suggested a subtle increase in serum P at the time of hCG injection without LH surge reduces the PR of women having oocyte retrievals for IVF; this study compared PRs of recipients in a shared oocyte program according to the donors' pre-hCG P level. There was no difference in viable PRs between recipients and donors when P < or = 1 ng/mL. The PR was similar for recipients when donors' P was > 1 ng/mL (12.7%). Donors with P > 1 ng/mL had the lowest PR: 7.2%. The data suggest that the adverse effect of higher serum P without LH surge may be on the endometrium rather than the oocyte.

Comparison of the cumulative probability of pregnancy after in vitro fertilization-embryo transfer by infertility factor and age.

OBJECTIVE: To compare the cumulative probability of pregnancy after multiple IVF cycles by age and cause of infertility. DESIGN: A prospective study was done in which patients were followed from the time they registered for their first IVF cycle until they achieved a clinical pregnancy, withdrew from treatment, or study was terminated. PATIENTS, SETTING, TREATMENTS: Infertile women undergoing IVF-ET at the Cooper Institute for In Vitro Fertilization were enrolled in this study if the luteal phase leuprolide acetate (LA) and hMG controlled ovarian hyperstimulation (COH) regimen was used. MAIN OUTCOME MEASURES: Clinical pregnancy, as determined by a positive beta-hCG level and ultrasonographic confirmation of a gestational sac, and delivery rates based on number of women with live births were compared by infertility factor and age. RESULTS: The 3-month cumulative probability of pregnancy based on life table analysis was 33% in women with tubal factor who were < or = 35 years of age, 25% in women with tubal factor who were > 35 years of age, 30% for women with multiple factors who were < or = 35 years of age, and 14% for women with multiple factors who were > 35 years of age. The rate for the older women with multiple factors was significantly lower than that for the other groups. The delivery rates were lower for the women with multiple factors than for women under 35 with tubal factor only. CONCLUSIONS: There is a significant effect of age and infertility factor on pregnancy and delivery rates. Physicians should consider these factors in evaluating their patients' prospects for success in IVF-ET.

Infusion of large particles of thiopental sodium during anesthesia induction.

STUDY OBJECTIVE: To determine the potential clinical significance of admixtures of thiopental sodium and acidic drugs, which are used during induction of general anesthesia and can cause the formation of particles of thiopental. DESIGN: Using an infusion setup similar to that used for a rapid-sequence induction of general anesthesia, injection of either pancuronium bromide or vecuronium bromide caused formation of particles of thiopental that were measured using a particle analyzer. The effects of delaying the injection of the muscle relaxant on particle formation and the effects of plasma on particle dissolution were studied. MEASUREMENTS AND MAIN RESULTS: The thiopental particles had a diameter of 17 to 39 microns, with a concentration of 15,000 to 20,000 particles/ml. Particle formation was prevented when a 30-second delay preceded administration of pancuronium or vecuronium following injection of thiopental. No particle formation was detected when succinylcholine was injected. Human plasma was far more effective than a crystalloid solution in dissolving thiopental particles. CONCLUSIONS: It is unlikely that clinically significant particles of thiopental will remain intact upon entering the bloodstream. However, mixing thiopental with pancuronium or vecuronium has the potential of disrupting intravenous access due to occlusion with particles.

Adverse effect of a homogeneous hyperechogenic endometrial sonographic pattern, despite adequate endometrial thickness on pregnancy rates following in-vitro fertilization.

We have previously presented data to show that in patients who had in-vitro fertilization (IVF)-embryo transfer using ovarian stimulation involving the luteal phase leuprolide acetate--human menopausal gonadotrophin (HMG) regimen, poor pregnancy results ensued if either the endometrial thickness was < 10 mm or a homogeneous hyperechogenic sonographic pattern was present immediately prior to taking a human chorionic gonadotrophin (HCG) injection. There were only 15 cases with this hyperechogenic type endometrium (and no pregnancies). The purpose of the present study was to evaluate the influence of a hyperechogenic endometrium when the endometrial thickness was > or = 10 mm, in a more extensive series, in women having IVF-embryo transfer using the same ovarian stimulation regimen. A total of 273 consecutive cycles, where endometrial thickness was > or = 10 mm, were evaluated (not including the 85 cycles previously reported). Of 22 patients with the hyperechogenic pattern, one achieved a chemical pregnancy (beta-HCG > 500 mIU/ml) and none achieved clinical pregnancies (ultrasound confirmation). In contrast, 67 of 251 (26.7%) patients conceived with other echo patterns (chi 2 analysis = 5.9, df = 1, P = 0.01). These data thus confirm, in a larger series, the negative influence of this type of echo pattern on subsequent pregnancy rates following the luteal phase leuprolide acetate--HMG ovarian stimulation regimen.

An evaluation of couples with failure of fertilization in vitro.

Attempts at in-vitro fertilization (IVF) may be used as a method of evaluating whether in a given couple, the inability of the sperm to fertilize the oocyte may be the cause of infertility. We evaluated all IVF patients in our practice who had at least one cycle with no fertilization to determine how often this was an isolated event or was repeated in multiple cycles; would poor semen quality be found as a frequent cause; and how well can a donor sperm or oocyte 'probe' uncover which of the two is the problem? Of 35 couples who used their own gametes exclusively, 30 (85.7%) had at least one cycle with zero fertilization; 42.5% of those failing to fertilize in cycle 1 and 35% of those failing in cycle 2 had a subnormal concentration of motile spermatozoa, morphology or hypo-osmotic swelling test scores. The pregnancy rate per cycle with both husband's and wife's gametes was only 2.3% (3/130), but was 8.3% for those using donor spermatozoa (3/36) and 18.2% (2/11) for donor oocytes. Thus, failing to fertilize in a given cycle does not necessarily predict failure to fertilize in a subsequent cycle, but does predict a poor fertility outcome unless donor gametes are used.