Urinary excretion profile of higenamine in females after oral administration of supplements - Doping scenario.

In 2017, higenamine was added to the World Antidoping Agency's (WADA) Prohibited list under group S3: beta-2 agonists and it is banned for athletes both in - and out of competition. Aim of this study was to characterize the urinary excretion profile of higenamine and its metabolite coclaurine after oral administration of multiple doses of higenamine capsules. For this purpose, an administration study including female basketball players was performed. For the detection of higenamine and cocalurine in the collected urine samples, a new, fast, and highly sensitive quantitative on-line SPE LC HRMS method was developed and validated. The method was applied for the quantification of higenamine and cocalurine in urine and their excretion pattern was defined. Results obtained show substantial inter-individual differences in the excretion profile of higenamine and coclaurine. For higenamine, half-lives were estimated to be between 4 and 27 h, and for coclaurine between 5 and 25 h. Furthermore, the data indicate that the elimination of coclaurine is rate-limited by its formation. Higenamine could be detected at a urine concentration above 10 ng/mL for at least 20 h after the last application for all study participants.

Treatment of Urethral Intercourse and Impact on Female Sexual Function.

BACKGROUND: Urethral intercourse is a very rare entity which usually presents as urinary incontinence during and after intercourse and is most commonly seen in patients with vaginal agenesis (Mayer-Rokitansky-Hauser Syndrome) or hypoplasia, or other rear vaginal anomalies. AIM: To evaluate management and outcomes for vaginal and urethral consequences of urethral intercourse, including urinary incontinence. METHODS: Between February 2006 and March 2021, 8 women aged from 17 to 22 years underwent genital and urethral reconstruction due to consequences of urethral sexual intercourse. Vaginal reconstruction included sigmoid vaginoplasty and introitoplasty with division of the vaginal septum in cases of vaginal agenesis (5 cases) and vaginal duplication (3 cases), respectively. Incontinence was treated by sling procedures in 5 women with longer history of urethral coitus and evident bladder neck prolapse. OUTCOMES: Sexual and psychosexual outcomes assessment was based on the Female Sexual Function Index and standardized questionnaires. RESULTS: Follow-up ranged from 9 to 188 months (mean 78 months). Good esthetical and functional results were achieved in all 8 women. All patients reported satisfactory sexual intercourse. All 5 incontinent women who had underwent sling procedure were continent. In one of 3 nontreated cases, additional sling treatment was indicated 6 months after vaginal reconstruction with satisfactory outcome. One patient with vaginal duplication reported a successful pregnancy with a Caesarean section delivery. CLINICAL IMPLICATIONS: Urinary incontinence with megalourethra in young women, along with the presence of Mullerian anomalies should raise suspicion of urethral coitus. Surgical treatment includes correction of vaginal anomalies and management of consequences. STRENGTHS AND LIMITATIONS: This study represents one of the largest series for urethral intercourse, with assessment of psychosexual outcome. The limitation is the lack of statistical analysis due to small sample size. CONCLUSION: Urethral intercourse is very rare, but it can cause severe consequences. It is important to recognize this occurrence and treat it by well-known vaginal or urethral reconstructive procedures. Djordjevic ML, Bizic M, Stojanovic B, et al. Treatment of Urethral Intercourse and Impact on Female Sexual Function. Sex Med 2022;10:100534.

AQbD-Oriented UHPLC/MS/MS Method Development for Glycopeptides Assessment in Pharmaceutical Forms.

Vancomycin and teicoplanin are glycopeptide antibacterials that inhibit the bacteria cell wall synthesis showing activity against gram-positive bacteria. Development of the sensitive method is of great importance for quality control of these drugs that are fermentation products. Modification of the fermentation conditions could cause the differences in the relative amount of the total substance or component, as it is the case with teicoplanin. The main objective of this study was development of the sensitive and effective ultra high performance liquid chromatography - tandem mass sprectrometry (UHPLC-MS/MS) method for simultaneous quantification of vancomycin, all six subcomponents of teicoplanin, and its pharmacopoeial impurity A in pharmaceutical forms. The scientific-based Quality by Design approach was implemented in the MS and UHPLC method development. Detection and quantification of analytes were carried out in positive electrospray ion mode by multiple reaction monitoring. Capillary voltage, cone voltage and collision energy were optimized by implementing experimental design methodology and optimal values for each fragment ion were obtained by performing experiments according to 'Rechtschaffen' design matrix. An ACQUITY CSH Phenyl-hexyl (2.1 × 50 mm, particle size 1.7 µm) column was chosen for the separation under the gradient elution mode with the mobile phase consisted of 0.1% formic acid in water (mobile phase A) and acetonitrile (mobile phase B). Optimal gradient elution parameters were achieved by applying 'Rechtschaffen' design too. Method operable design regions were constructed for investigated MS and chromatographic parameters. The method was fully validated, and its applicability was confirmed throughout the ability to follow the behavior of vancomycin and teicoplanin under stress conditions.

Tribocorrosion of a CoCrMo alloy sliding against articular cartilage and the impact of metal ion release on chondrocytes.

Partial knee replacement and hemiarthroplasty are some of the orthopedic procedures resulting in a metal on cartilage interface. As metal implant material, CoCrMo based alloys are commonly used. The aim of the present study is to assess the role of biotribocorrosion on the CoCrMo-cartilage interface with an emphasis on metal release during sliding contact. The biotribocorrosion experiments were performed under controlled electrochemical conditions using a floating cell with a three electrode set up coupled to a microtribometer. Throughout the experiment the coefficient of friction and the open circuit potential were monitored. Analyses of the electrolyte after the experiment show that metal release can occur during sliding contact of CoCrMo alloy against articular cartilage despite the extraordinary low coefficient of friction measured. Metal release is attributed to changes in passive layer caused at the onset of sliding. The released metal was found to be forming compounds with potential cytotoxicity. Since the presence of metal ions in the cartilage matrix can potentially lead to cell apoptosis, the metabolic activity of human osteoarthritic chondrocytes (2D-cultures) was investigated in the presence of phosphate buffered saline containing metal ions using XTT-assay. The experiments indicate that critical concentrations of Co ions lead to a significant decrease in chondrocyte metabolic activity. Therefore, biotribocorrosion is a mechanism that can occur in partial replacements and lead to chondrocyte apoptosis thus playing a role in the observed accelerated degradation of the remaining cartilage tissue after the mentioned orthopedic procedures. STATEMENT OF SIGNIFICANCE: Partial replacements provide an alternative to total joint replacements. This procedure is less invasive, allows a faster rehabilitation and provides a better function of the joint. However, the remaining native cartilage experiences accelerated degradation when in contact with metallic implant components. This work investigates the role of tribocorrosion at the metal-cartilage interface during sliding. Tribocorrosion is a degradation process that can alter significantly the wear rates experienced by metallic implants and lead to the release of metal ions and particles. The released metal can form compounds with potential cytotoxicity on cartilage tissue. The knowledge gained in this work will serve to understand the mechanisms behind the failure of partial replacements and develop future biomaterials with an enhanced lifetime.

Hysterectomy with Bilateral Salpingo-Oophorectomy in Female-to-Male Gender Affirmation Surgery: Comparison of Two Methods.

INTRODUCTION: The optimal route for hysterectomy with bilateral salpingo-oophorectomy in female-to-male gender affirmation surgery is still under debate, due to the quite limited and inconsistent published data. The aim of this study is to present and compare the results of vaginal and laparoscopic hysterectomy as part of gender affirmation surgery in female-to-male transsexuals. MATERIALS AND METHODS: Between 2012 and 2017, 124 female-to-male transsexuals, aged 18-43 years (mean age: 28.5), underwent hysterectomy with bilateral salpingo-oophorectomy, followed by colpocleisis and gender affirmation surgery. Transvaginal and laparoscopic hysterectomy were performed in 92 and 32 patients, respectively. Standard outcome measures (types and rates of complications, operative time, blood loss, and postoperative hospital stay) were used to compare the two groups of patients. RESULTS: The mean follow-up was 41 months (ranged from 6 to 65 months). The duration of transvaginal approach was significantly shorter (51 minutes compared to 76 minutes, p < 0.001). The total complication rates (less than 3%), reoperation rates (0%), blood loss, and postoperative hospital stays (4.3 days compared to 4.5 days) showed no statistical difference. CONCLUSIONS: Both approaches are safe, with minimal complications. However, we prefer transvaginal hysterectomy due to its shorter operative time, cost-effectiveness, and simpler continuation with one-stage female-to-male gender affirmation surgery.

Determination of higenamine in dietary supplements by UHPLC/MS/MS method.

From 1st January 2017 higenamine was added on the WADA (World Anti-doping Agency) Prohibited list under S3 group beta-2 agonists as at all times banned substance for the athletes. The main origine of higenamine (or norcoclaurine) are different plants including Nandina domestica, Aconitum carmichaelii, Asarum heterotropioides, Galium divaricatum, Annona squamosa, Nelumbo nucifera etc. Higenamine main use is related to weight loss and it could be found (un)labeled in different dietary supplements. The objective of this study was development of sensitive and reliable UHPLC/MS/MS method for determination of higenamine in various dietary supplement samples. In order to obtain high method sensitivity, hydrophilic interaction liquid chromatography (HILIC) mode was applied. Separation was carried out on UHPLC Acquity BEH HILIC analytical column (2.1mm×100mm, 1.7µm particle size). Mobile phase consisted of 0.1% formic acid in water and acetonitrile, respectively, was mixed in ratio of 30:70, v/v. Flow rate was set at 0.2mLmin-1. Quercetin was used as an internal standard. ESI (+) source ionization mode using multi reaction monitoring (MRM) mode was utilized and three ion transitions of higenamine were followed 272.08→107.01, 272.08→161.07 and 272.08→77.08. Developed method was fully validated and applied for identification and quantification of higenamine in different dietary supplements. According to the results, the most of investigated supplements were free of higenamine, and on the other hand, presence of higenamine was confirmed in some samples while it was not declared on the label.

Effect of physical effectiveness on digestibility of ration for cows in early lactation.

A study was conducted to investigate the effects of a diet particle size on nutrient digestibility in cows in early lactation. Treatments were diets with forage to concentrate ratio 43:57% in diet dry matter, with four different physically effective fibres (peNDF) content based on different cut length of corn silage and alfalfa haylage. The physical effectiveness factors (pef) and peNDF content of TMRs (total mixed ration) were determined using Penn State Particle Separator (PSPS) with two (pef8.0 , peNDF8.0) or three (pef1.18, peNDF1.18) sieves. The reducing of cut length of forages and particle size of diets did not affect on dry matter intake, while decreased peNDF intake by 16.34 and 8.83%, for peNDF8.0 and peNDF1.18 respectively. Apparent total tract digestibility of the nutrients was measured using two indicators: acid insoluble ash (AIA) and acid detergent insoluble lignin (ADL). Decreasing of forages cut length significantly increased apparent total tract digestibility of neutral detergent fibre (NDF) from 48.39% to 53.84% and from 53.9% to 58.66%, of crude protein from 73.96% to 79.24% and from 71.56% to 77.90%, with contemporary decreasing of non-fibre carbohydrate from 90.89% to 84.81% and from 91.99% to 86.80%, with AIA or ADL as indicator respectively. Dietary value of net energy for lactation (NEL) and energy intake was not affected by the peNDF content of the diet.

Systematical approach in evaluation of LC method for determination of raloxifene hydrochloride and its impurities employing experimental design.

Method validation presents a detailed investigation of analytical method and provision of the evidence that the method, when correctly applied, produces results that fit to the purpose. In order to achieve the method validation scope efficiently, experimental design presents a very useful tool. The greatest benefits of such approach could be seen in robustness testing through the provision of very useful data about the control of the chromatographic system during the routine application. In this paper, robustness testing of the LC method proposed for the determination of raloxifene hydrochloride and its four impurities was done employing Plackett-Burman design. Applying this design, the effect of five real factors (acetonitrile content, sodium dodecyl sulfate content, column temperature, pH of the mobile phase and flow rate) on the corresponding resolution factors was investigated through twelve experiments. Furthermore, the insignificance intervals for significant factors were calculated and the parameters for system suitability tests were defined. Eventually, the other validation parameters were tested and the effectiveness of the proposed analytical method with a high degree of accuracy was confirmed.

Geometric hysteresis of alveolated ductal architecture.

Low Reynolds number airflow in the pulmonary acinus and aerosol particle kinetics therein are significantly conditioned by the nature of the tidal motion of alveolar duct geometry. At least two components of the ductal structure are known to exhibit stress-strain hysteresis: smooth muscle within the alveolar entrance rings, and surfactant at the air-tissue interface. We hypothesize that the geometric hysteresis of the alveolar duct is largely determined by the interaction of the amount of smooth muscle and connective tissue in ductal rings, septal tissue properties, and surface tension-surface area characteristics of surfactant. To test this hypothesis, we have extended the well-known structural model of the alveolar duct by Wilson and Bachofen (1982, "A Model for Mechanical Structure of the Alveolar Duct," J. Appl. Physiol. 52(4), pp. 1064-1070) by adding realistic elastic and hysteretic properties of (1) the alveolar entrance ring, (2) septal tissue, and (3) surfactant. With realistic values for tissue and surface properties, we conclude that: (1) there is a significant, and underappreciated, amount of geometric hysteresis in alveolar ductal architecture; and (2) the contribution of smooth muscle and surfactant to geometric hysteresis are of opposite senses, tending toward cancellation. Quantitatively, the geometric hysteresis found experimentally by Miki et al. (1993, "Geometric Hysteresis in Pulmonary Surface-to-Volume Ratio during Tidal Breathing," J. Appl. Physiol. 75(4), pp. 1630-1636) is consistent with little or no smooth muscle tone in anesthetized rabbits in control conditions, and with substantial smooth muscle activation following methacholine challenge. The observed local hysteretic boundary motion of the acinar duct would result in irreversible acinar flow fields, which might be important mechanistic contributors to aerosol mixing and deposition deep in the lung.

Advancement in optimization tactic achieved by newly developed chromatographic response function: application to LC separation of raloxifene and its impurities.

In this paper a new chromatographic response function (CRF) is designed and proposed for utilization in the optimization strategies. The function capability to represent the overall quality of a experimentally obtained chromatograms was compared to the other two objective functions and proved to give more accurate and reliable results. The new CRF has improved concept of separation and time term estimation. It reflects all important defects of the chromatogram such as the appearance of asymmetrical or overlapping peaks and prolonged elution time and allows the appropriate weighting of each of them. The LC separation of raloxifene and its four impurities was evaluated through the central composite design experimental plan choosing the new CRF to be the only output of the system. The function demonstrated the ability to judge the impact of the complex interactions of the selected chromatographic parameters (acetonitrile content in the mobile phase, sodium dodecyl sulfate concentration in the water phase, pH of the mobile phase and column temperature) on the mixture behavior and led to the determination of the optimal separation conditions. The newly developed CRF proved to have the advanced performances and it presents the important step forward in the optimization of the chromatographic separation.

Artificial neural networks in analysis of indinavir and its degradation products retention.

Artificial neural networks (ANN) are biologically inspired computer programs designed to simulate the way in which the human brain processes the information. In the past few years, coupling of experimental design (ED) and ANN became useful tool in the method optimization. This paper presents the application of ED-ANN in analysis of chromatographic behavior of indinavir and its degradation products. According to preliminary study, full factorial design 2(4) was chosen to set input variables for network training. Experimental data (inputs) and results for retention factors from experiments (outputs) were used to train the ANN with aim to define correlation among variables. For networks training multi-layer perceptron (MLP) with back propagation (BP) algorithm was used. Network with the lowest root mean square (RMS) had 4-8-3 topology. Predicted data were in good agreement with experimental data (correlation was higher than 0.9713 for training set). Regression statistics confirmed good ability of trained network to predict compounds retention.

Chemometrical evaluation of ropinirole and its impurity's chromatographic behavior.

The aim of this study was the chemometrical evaluation of ropinirole and its impurity's (4-[2-(dipropylamino)ethyl]-1H-indol-2,3-dione) chromatographic behavior in systematic and the most efficient way. For that purpose, as very descriptive, response surface designs are most preferable. Face-centered central composite design (CCD) with 2(3) full factorial design, +/-1 star design and four replication in central point was applied for a response surface study, in order to examine in depth the effects of the most important factors. Factors-independent variables (acetonitrile content, pH of the mobile phase and concentration of sodium heptane sulfonate in water phase) were extracted from the preliminary study and as dependent variables five responses (retention factor of ropinirole, retentin factor of its impurity, resolution, symmetry of ropinirole peak and symmetry of impurity peak) were selected. For the improvement of method development and optimization step, Derringer's desirability function was applied to simultaneously optimize the five chosen responses. The procedure allowed deduction of optimal conditions and the predicted optimum was acetonitrile-5mM of sodium heptane sulfonate (21.6:78.4, v/v), pH of the mobile phase adjusted at 2.0 with ortho phosphoric acid. By calculating global desirability's determination coefficients (R(D)(2)), as well as by the visual inspection of 3D graphs for global desirability, robustness of the proposed method was also estimated.

Modeling of muscle fatigue using Hill's model.

A new model incorporating muscle fatigue has been developed to predict the effect of muscle fatigue on the force-time relationship of skeletal muscle by using the PAK-program. Differential equations in the incremental form have been implemented into Hill's muscle model. In order to describe the effect of muscle fatigue and recovery on skeletal muscle behaviors, a set of equations in terms of three phenomenological parameters which are a fatigue curve under sustained maximal activation, a recovery curve and an endurance function were developed. With reference to existing models and experimental results, the input parameters for fatigue curve under sustained maximal activation and endurance function were determined. The model has been investigated under an isometric condition. The effects of different shapes of the recovery curves have also been considered in this model. Validation of the model has been performed by comparing the predicted results with the experimental data from an existing literature.

Ia+T10+ blast cells in acute lymphoblastic leukaemia are precursors of T+ and B+ cells.

Leukaemic blasts isolated from peripheral blood of two patients with non-T, non-B ALL were cultivated for 10-13 days and cell markers were assayed on day 0 and during culture period. Using a panel of cell markers it was demonstrated that the populations of ALL blasts of both patients were composed of T10+Ia+ and "null" blasts. These blasts in the culture differentiated into three cell lines: T and B lymphocytes and myelomonocytic cells. The results thus obtained suggested that within the population of ALL blasts of these two patients there are progenitors of lymphoid and myelomonocytic cells ("null" blasts) and common precursors of T and B lymphocytes (T10+Ia+ blasts). In vitro differentiation of ALL blasts was not affected by LPS and CTPE.

[Reconstruction of the common bile duct with a free venous autograft (experimental work - preliminary communication)]. / Rekonstrukcija duktus holedohusa slobodnim autovenskim graftom (eksperimentalni rad - preliminarno saopstenje).

In seven dogs, the authors performed a resection of the supraduodenal part of the main bile duct, 15-2 cm. in length. The arising defect was bridged over with a free autovenous graft taken from the exterior jugular vein (T-T anastomosis), without using a temporary or permanent prosthesis. The animals were followed-up 60 days and the sacrificed. Three dogs perished: the first one on the third day due to intoxication, the second dog on the thirteenth day because of anastomosis' disruption and consequent biliary peritonitis, and the third one on the eighteenth day of unknown causes. In the immediate postoperative course the values of the bilirubin and alkaline phosphatase were increased while those of transaminase were not significantly altered. These values began to normalize during the third week. An intravenous biliography was made on the sixtieth day showing a normal filling of the gallbladder, with an orderly visualization of hepato-choledochus and its correct transit, but with stenosis at the distal anastomosis. Macroscopically, the graft appeared slightly enlarged in length and diameter due to the mentioned stenosis. Histologically, the graft took on an appearance similar to the main bile duct, while its endothelium was completely replaced by biliary epithelium. An answer about the viability of this autovenous graft to serve as a substituent of a normal common bile duct is to be expected with further experimental work.