Excretion Time Research Articles

African swine fever: virus carriage and the role of surviving wild boar in the persistence and spread of infection (review)

The panzootic of African swine fever (ASF) in Europe and Asia caused by viruses of genotypes I and II has led to a comprehensive scientific study of the course of this infection. Researchers have noted that a certain proportion of diseased animals survive. Detection of seropositive (antibody-positive) animals in ASF-affected countries of Europe was especially noticeable in wild boar populations. The role of seropositive animals ASF survivors in the persistence and spread of infection in the population has been long and controversially debated, as they may potentially become persistently infected, acting as virus carriers. The aim of this review was to summarize the current scientific and experimental results on the chronic course of ASF in Eurasian wild boar, virus carriage and spread of infection from surviving seropositive animals. In Eurasia different forms of ASF disease are currently observed in susceptible animals: peracute, acute, subacute, less frequently chronic and asymptomatic; the latter are found to be caused by circulating ASF viruses (ASFV) of reduced virulence. Two types of survivors animals are distinguished: 1) animals that develop persistent infection with periodic viremia and signs of subacute to chronic course; 2) animals that recover completely and clear of infection. Long-term persistence and complete elimination of the virus have been observed in survivors: in the last ten years, the time of virus excretion has been experimentally determined to generally range from 35 to 99 days. Survivors of the 1st type may play a role in the spread of the ASFV due to periodic viremia. Seropositive survived animals of the 2nd type due to their small numbers are not considered by some researchers to play a significant epidemiological role in the persistence of ASFV in wild boar populations. Experimental transmission of ASFV from surviving wild boars has been studied to date to a limited extent, such research should be continued. The knowledge gained in these areas will improve the understanding of the current situation of ASF in wildlife.

Metabolite profiling of Shenhua compound in rats and pharmacokinetics study of four bioactive compounds with liquid chromatography combined with electrospray ionization tandem mass spectrometry.

Shenhua compound, composed of ginseng, hawthorn and sophora flower, has been shown to improve hyperlipidemia. However, the main ingredients, their metabolic pathways in vivo, and pharmacokinetic characteristics. In this study, ultra-high-performance liquid chromatography coupled with electrospray ionization quadruple time-of-flight mass spectrometry (UHPLC-Q-TOF) was used to qualitatively analyze the main ingredients in ethanol extract of Shenhua compound and to investigate the metabolites in serum, bile, feces, and urine of rats following oral administration. The pharmacokinetics of ginsenoside Rg1, ginsenoside Re, ginsenoside Ro and rutin in rats were analyzed using triple-quadrupole liquid chromatography combined with electrospray ionization mass spectrometry (QQQ-LC/MS). The results indicated that 48 compounds were present in Shenhua compound, including saponins, flavonoids and organic acids. Metabolites were comprehensively analyzed after oral administration of Shenhua compound, and 24 prototype ingredients and 92 metabolite ingredients were identified or characterized. By analyzing the pharmacokinetic parameters of ginsenoside Rg1, ginsenoside Re, ginsenoside Ro and rutin from 0 to 72 h after oral administration of various dose of Shenhua compound, it was observed that the concentration of ginsenosides in blood remained below 2 ng∙mL1, but the metabolic excretion time was prolonged. Meanwhile, the blood concentration of rutin was significantly higher than ginsenosides and showed a double absorption peak. In conclusion, this study provides valuable insights into compound ingredients metabolism regularities in vivo and pharmacokinetics after oral administration of Shenhua compound.

Effects of endurance exercise under hypoxic conditions on the gastric emptying rate and intestinal cell damage.

The present study examined the effects of gastric emptying rate and intestinal cell damage following a single session of endurance exercise under "hypoxic" or "normoxic" conditions at the same relative intensity. Eleven healthy males performed two trials on different days, consisting of a 60min run on a treadmill at 70% maximal running velocity (vMax) while inspiring hypoxic (FiO2: 14.5%; HYP) or normoxic air (FiO2: 20.9%; NOR). The average running velocity was 11.4 ± 0.7km/h in NOR and 10.8 ± 0.5km/h in HYP, respectively. Venous blood samples were collected to evaluate plasma intestinal fatty acid binding protein (I-FABP) as an indicator of exercise-induced intestinal cell damage. The gastric emptying rate was determined by the 13C-sodium acetate breath test. Running velocities at 70% vMax and arterial oxygen saturation were significantly lower under HYP than NOR (p < 0.001). Peak heart rate and rating of perceived exertion during exercise did not differ significantly between the trials. Maximum 13C excretion time (an indication of the gastric emptying rate) was significantly delayed in the HYP (NOR: 38.5 ± 5.0min, HYP: 45.5 ± 9.6min; p = 0.010). Furthermore, the score of nausea increased slightly, but increased significantly after exercise only in the HYP (p = 0.04). However, exercise-induced changes in plasma I-FABP, adrenaline, and noradrenaline concentrations did not differ significantly between the two trials. These results suggest that endurance exercise under hypoxic conditions impairs digestive function in the stomach compared to exercise under normoxic conditions performed at the same relative intensity.

Effect of feeding time and time of harvest of fresh pasture on urinary and faecal nitrogen excretion patterns in sheep

ABSTRACT Under New Zealand grazing systems, a high proportion of the nitrogen consumed by ruminants is excreted. This experiment examined the effect of feeding morning and afternoon harvested annual ryegrass, fed shortly after harvest, on the timing of nitrogen excretion in one-year-old sheep during winter. Dry matter and water-soluble carbohydrate (WSC) content increased in the afternoon-harvested ryegrass. This resulted in increased protein and WSC intakes and a decrease in nitrogen loss in the sheep fed in the afternoon with afternoon-harvested grass. The pattern of urinary nitrogen excretion was related to feeding time with peak urinary urea excretion in the period 4–8 hours after feeding, with over 60% of the daily urinary urea excreted in the 12 hours after the feed was offered. This study suggests that shifting stock onto new pasture breaks in the afternoon could reduce nitrous oxide production by increasing the efficiency of nitrogen utilisation in animals and reducing the total amount of nitrogen excreted. It may be possible to utilise the potential diurnal differences in pasture composition to reduce nitrogen losses. This practise could also improve animal performance as well as change the timing of nitrogen excretion.

Natural compound Sanggenon C inhibits porcine reproductive and respiratory syndrome virus replication in piglets

Porcine reproductive and respiratory syndrome virus is one of the main pathogens threatening the global pig industry, and there is still a lack of effective therapeutic drugs. Sanggenon C is a flavanone Diels-Alder adduct compound extracted from the root bark of the mulberry genus, which has blood pressure-reducing, anti-atherosclerotic, anti-oxidative, and anti-inflammatory effects. In our previous study, Sanggenon C was confirmed to significantly inhibit PRRSV replication in vitro. However, its antiviral potential to inhibit PRRSV infection in vivo has not been evaluated in piglets. Here, the antiviral effect of Sanggenon C was evaluated in PRRSV-challenged piglets based on assessments of rectal temperature, viral load, pathological changes of lung tissue and secretion of inflammatory cytokines. The results showed that Sanggenon C treatment relieved the clinical symptoms, reduced the viral loads in the lungs and bloods, alleviated the pathological damage of lung tissue, decreased the secretion of inflammatory cytokines, and shorten the excretion time of virus from the oral and nasal secretions and feces of piglets after PRRSV infection. The results indicated that Sanggenon C is a promising anti-PRRSV drug, which provides a new strategy for the prevention and control of PRRS in clinical practice.

Enhancing Physicochemical Characteristics of Donkey Meat through High‐Voltage Electrostatic Stimulation during Acid Excretion

To investigate the impact of high‐voltage electrostatic stimulation on the quality of donkey meat, ten 30‐month‐old Dezhou donkeys (each weighing 250 ± 50 kg) were selected. Following slaughter, longissimus dorsi (LD) samples from both left and right carcasses were collected. The control group refrigerated the right muscle in a 0–4°C refrigerator for normal acid excretion. Meanwhile, the high‐voltage electrostatic group subjected the left muscle to high‐voltage electrostatic stimulation (30000 V) for 60 s before refrigerating it in the same conditions for acid excretion. The study showed that at the 1st, 6th, and 24th h of acid excretion, the high‐voltage electrostatic group exhibited significantly lower pH levels than the control group (P&lt;0.05). Furthermore, a∗ values in the high‐voltage electrostatic group showed an increasing trend relative to the control group at the 6th and 24th h of acid excretion (P = 0.0751, P = 0.0860). With increasing acid excretion time, both treatment groups displayed increased PV and TBARS values. At 48 and 72 h, the PV values in the high‐voltage electrostatic group were significantly lower than those in the control group (P &lt; 0.05). As acid excretion time increased, T‐AOC, SOD, and GSH‐Px in both groups decreased. However, the high‐voltage electrostatic group consistently exhibited higher antioxidant enzyme activity than the control group at all time points. SOD and GSH‐Px activity in the high‐voltage electrostatic group showed an increasing trend at 48 h of acid excretion (P = 0.0838, P = 0.0860). At 72nd h of acid excretion, the shear force of donkey meat in the high‐voltage electrostatic group was significantly lower than that in the ordinary treatment group (P &lt; 0.05). With increasing acid excretion time, the MFI of both treatment groups gradually increased. In particular, at the 1st and 48th h of acid excretion, the MFI of the high‐voltage electrostatic group was significantly higher than that of the ordinary treatment group (P &lt; 0.05). Following high‐voltage electrostatic stimulation, ultrastructure images of donkey meat at 6 and 72 h revealed numerous contractions and stretching bands. Therefore, this stimulation significantly improved the tenderness of donkey meat, reduced the oxidation rate of donkey meat, and improved the quality of donkey meat.

Food chain-mediated variation in excretion times of microplastics: Unraveling the interactions with plasticizers

Microplastics are global environmental pollutants. Further, there has been growing concern regarding the contamination of plasticizers. In this study, we first found that Daphnia magna ingests microplastics of red, blue, yellow, green, black, white, and clear colors. Next, using D. magna (crustacean) and zebrafish (fish), we investigated whether the time required to expel microplastics varies when ingestion occurs through a food-chain. As a results, we found that microplastics excretion occurs within 48 h and 24 h for direct ingestion and ingestion through food chain (via D. magna), respectively. Lastly, we examined whether acute toxicity values varied when D. magna was exposed to a combination of plasticizers and microplastics, compared to exposure to plasticizers alone, and found that there were no differences in the acute toxicity values. This study provides important data for investigating the impacts of microplastics on ecosystems and is expected to contribute to future ecological risk assessments of microplastics.

Use of glucarpidase (carboxypeptidase-G2) in pediatric cancer patients: 11-year experience of a tertiary center.

Methotrexate is an essential drug in the treatment of childhood cancer that is not exempt from toxicities. Glucarpidase is a drug used to reduce the toxic concentration of plasma methotrexate in patients with delayed elimination or at risk of toxicity. We describe the characteristics of a cohort of pediatric patients that received glucarpidase and analyze its role in the treatment of toxicity induced by high doses of methotrexate (HDMTX). Retrospective observational study of all pediatric cancer patients who received glucarpidase between 2012 and 2022 at a single center. Fifteen patients were treated with a single dose of glucarpidase, eleven of them presented with acute lymphoblastic leukemia and received HDMTX at 5g/m2 in 24-hour infusion. In eight patients, glucarpidase was administered during the first cycle of HDMTX. The indication in thirteen cases was acute renal failure with delayed elimination of plasma methotrexate. The median maximum creatinine was 1.22mg/dl (0.682.01mg/dl), with a median increase over its baseline level of 313%. All patients normalized renal function after glucarpidase administration, with a median methotrexate excretion time of 193 hours (42-312hours). No grade ≥2 adverse events derived from carboxypeptidase administration. Eleven patients received new doses of HDMTX in subsequent cycles, without new episodes of serious toxicity. The use of glucarpidase is effective and safe in the treatment of acute renal failure and methotrexate elimination delay in pediatric cancer patients. Further HDMTX doses may be prescribed without additional toxicities.

How important is food structure when cats eat mice?

Feeding whole prey to felids has shown to benefit their gastrointestinal health. Whether this effect is caused by the chemical or physical nature of whole prey is unknown. Fifteen domestic cats, as a model for strict carnivores, were either fed minced mice (MM) or whole mice (WM), to determine the effect of food structure on digestibility, mean urinary excretion time (MUET) of 15N, intestinal microbial activity and fermentation products. Faeces samples were collected after feeding all cats a commercially available extruded diet (EXT) for 10 d before feeding for 19 d the MM and WM diets with faeces and urine collected from day 11 to 15. Samples for microbiota composition and determination of MUET were obtained from day 16 to 19. The physical structure of the mice diet (minced or not) did not affect large intestinal fermentation as total SCFA and branched-chain fatty acid (BCFA), and most biogenic amine (BA) concentrations were not different (P > 0·10). When changing from EXT to the mice diets, the microbial community composition shifted from a carbolytic (Prevotellaceae) to proteolytic (Fusobacteriaceae) profile and led to a reduced faecal acetic to propionic acid ratio, SCFA, total BCFA (P < 0·001), NH3 (P = 0·04), total BA (P < 0·001) and para-cresol (P = 0·08). The results of this study indicate that food structure within a whole-prey diet is less important than the overall diet type, with major shifts in microbiome and decrease in potentially harmful fermentation products when diet changes from extruded to mice. This urges for careful consideration of the consequences of prey-based diets for gut health in cats.

Effect of deep needling "Tianshu" (ST25) on colonic motility in rats with slow transit constipation

To observe the effect of acupuncture of "Tianshu"(ST25) at different depths on colonic transportation function, expressions of colonic substance P (SP) and interstitial cells of Cajal (ICC) in rats with slow transit constipation (STC), so as to explore its mechanisms underlying improvement of STC．. Fifty male Wistar rats were selected and randomly divided into control，STC model，conventional acupuncture，deep needling group 1 and deep needling group 2 groups，with 10 rats in each group．The STC model was established by gavage of 1 mg/mL compound diphenoxylate suspension (10 mg/kg), once every other day for 21 days, and rats of the control group were given the same dose of distilled water by gavage．EA (2 Hz, 2 mA) was applied to "Tianshu"(ST25), with the acupuncture needle inserted to a depth of 3 mm for rats of the conventional acupuncture group, 4.5 mm for those of deep needling group 1, and 10 mm for those of the deep needling group 2. The acupuncture needle was twirled for 1 min, then retained for 15 min each time, once a day for 15 consecutive days．Following modeling, rats of the 4 groups and the control group received gavage of active carbon 2 mL (100 g/L) for observing the excretion time of the first black stool grain to assess the intestinal transit function. The colonic myoelectric activities (frequency and amplitude) were recorded by using BIOPAC multichannel physiograph. The immunoactivity of SP and c-kit (a transmembrane protein kinase for identification of ICC) of colonic musculature was detected by immunohistochemistry. Compared with the control group，the time of excretion of the first black stool grain, and the amplitude of colonic electromyogram (EMG) were significantly increased (P<0.01), while the frequency of EMG, expressions of SP and c-kit (ICC) were significantly decreased in the model group (P<0.01). In contrast to the model group, both deep needling group 1 and 2 had a decrease in the time of excretion of the first black stool grain, and amplitude of intestinal EMG, and an increase of frequency of intestinal EMG, and immunoactivity of SP and c-kit (P<0.01). The effect of deep needling 2 is superior to that of deep needling 1 in reducing the time of excretion of the first black stool grain (P<0.05), lowering the amplitude of EMG of the gut smooth muscle (P<0.05) and in increasing the frequency of EMG (P<0.05) and the expressions of SP and c-kit (P<0.05). No significant changes were found in the levels of frequency and amplitude of EMG, and expressions of SP and c-kit after routine needling in comparison with the model group (P>0.05), except the excretion time of the first black stool grain (P<0.05). Deep needling at ST25 at depth of 4.5 mm and 10 mm，especially at depth of 10 mm，has a significant effect in promoting gut motility to ameliorate constipation in rats with STC, which may be related to its function in up-regulating the expressions of SP and ICC activity.

Establishment and identification of an animal model of Hirschsprung disease in suckling mice

BackgroundHirschsprung disease (HSCR) is a congenital intestinal malformation. Previous HSCR animal model needs invasive operation on adult animal. The aim of this study is to establish an early-onset animal model which is consistent with the clinical manifestation of HSCR patients.MethodsThe neonatal mice were randomly divided into the benzalkonium chloride (BAC) group, treated with BAC via enema, and the control group, treated with saline. Weight changes, excretion time of carmine, CT scan, hematoxylin-eosin staining and immunofluorescence staining were used to evaluate the effect of the model. Differentially expressed genes (DEGs) in the HSCR mice were analyzed by using DAVID 6.8 database and compared with DEGs from HSCR patients.ResultsThe weight of mice was lower and the excretion time of carmine was longer in the BAC group. Moreover, distal colon stenosis and proximal colon enlargement appeared in the BAC group. Neurons in the distal colon decreased significantly after 4 weeks of BAC treatment and almost disappeared completely after 12 weeks. Transcriptome profiling of the mouse model and HSCR patients is similar in terms of altered gene expression.ConclusionsAn economical and reliable HSCR animal model which has similar clinical characteristics to HSCR patients was successfully established.ImpactThe animal model of Hirschsprung disease was first established in BALB/c mice.This model is an animal model of early-onset HSCR that is easy to operate and consistent with clinical manifestations.Transcriptome profiling of the mouse model and HSCR patients is similar in terms of altered gene expression.

Pharmacological profile, phase I metabolism, and excretion time profile of the new synthetic cathinone 3,4-Pr-PipVP.

1-(2,3-Dihydro-1H-inden-5-yl)-2-(piperidin-1-yl)pentan-1-one (3,4-Pr-PipVP), a novel synthetic cathinone (SCat), was first identified in 2022 in Germany. The product was marketed as 1-(bicyclo[4.2.0]octa-1,3,5-trien-3-yl)-2-(pyrrolidin-1-yl)pentan-1-one (3,4-EtPV), a substance not covered by the German New Psychoactive Substances Act (NpSG). Although originally intended to be an exploratory new synthetic cathinone containing the novel bicyclo[4.2.0]octatrienyl function, the compound was subsequently confirmed to contain an indanyl ring system scheduled under generic legislation like the NpSG. However, it is one of only a few marketed SCats carrying a piperidine ring. Inhibition experiments involving norepinephrine, dopamine, and serotonin transporters showed that 3,4-Pr-PipVP was a low potency blocker at all three monoamine transporters compared to related substances such as MDPV. Additionally, pharmacokinetic data were collected from pooled human liver microsomes incubations and from the analysis of authentic urine samples received after oral administration of 5mg 3,4-Pr-PipVP hydrochloride. Phase I metabolites were tentatively identified in vitro and in vivo using liquid chromatography-time-of-flight mass spectrometry. Main metabolites were formed by metabolic reduction of the carbonyl function with and without additional hydroxylations at the propylene bridge of the molecule. Keto-reduced H2 -3,4-Pr-PipVP and H2 -piperidine-OH-3,4-Pr-PipVP as well as aryl-OH-3,4-Pr-PipVP, and indanyl-OH-piperidine-OH-3,4-Pr-PipVP are suggested as most suitable biomarkers for the detection of 3,4-Pr-PipVP since they were detected for much longer than the parent compound. 3,4-Pr-PipVP could be detected for up to 21 h whereas its metabolites were detectable for up to about 4 days.

Welfare impacts associated with using aversive geofencing devices on captive Asian elephants

Animal-borne aversive geofencing devices (AGDs, or satellite-linked shock collars) are commercially available and used on livestock to restrict their movement within a virtual boundary. This technology has potential application as a human-wildlife conflict mitigation tool, where problem animals might be conditioned to avoid human-dominated habitats by associating an audio warning with a subsequent electric shock, which is delivered if the audio warning is ignored. Ensuring that high standards of animal welfare are maintained when implementing such tools is important for acquiring manager and community acceptance of such approaches. We conducted two pilot experiments with eight captive Asian elephants using mild electric shocks from a modified dog-training collar fitted around the neck, as part of an ongoing effort to develop AGDs suitable for mitigating human-elephant conflict. As part of these experiments, we assessed elephants' behavioural and physiological stress before, during and after the experiments. During the experiments elephants wore collars for up to nine consecutive days and received a small number of electric shocks on 1–3 consecutive days. Bootstrapped principal component analysis showed that daily activity budgets of individual elephants on experiment days were not different from the pre-experiment days. Generalised linear mixed-effects model (GLMM) showed that anxiety/stress behaviours increased on the first day of acclimatising to the collar and on testing days (i.e. days they received shocks) of the first experiment, but not during the second experiment relative to pre-experiment days. Analysis of faecal cortisol metabolite (FCM) concentrations using GLMM showed that FCM concentrations were higher in samples collected ∼24 hrs and ∼48 hrs after testing days compared to baseline levels as expected given the lag time for excretion of cortisol metabolites. These elevated anxiety/stress behaviours and FCM concentrations returned to baseline levels shortly after the experiment. Therefore, we conclude that AGDs did not produce lasting behavioural or physiological stress effects in elephants during this short-term study but recommend further studies with a larger sample of elephants to confirm the transferability of these findings.

LC-HRMS based metabolomics workflow: an alternative strategy for metabolite identification in the anti-doping field.

The proposed metabolomic workflow, based on coupling high-resolution mass spectrometry with computational tools, can be an alternative strategy for metabolite detection and identification. This approach allows the extension of the investigation field to chemically different compounds, maximizing the information obtainable from the data and minimizing the time and resources required. Urine samples were collected from 5 healthy volunteers before and after oral administration of 3β-hydroxyandrost-5-ene-7,17-dione as a model compound and defining three excretion time intervals. Raw data were acquired in both positive and negative ionization modes using an Agilent Technologies 1290 Infinity II series HPLC coupled to a 6545 Accurate-Mass Quadrupole Time-of-Flight. They were then processed to align peak retention times with the same accurate mass, and the resulting data matrix was subjected to multivariate analysis. Multivariate analysis (PCA and PLS-DA models) demonstrated high similarity between samples belonging to the same collection time interval and clear discrimination between different excretion intervals. The blank and long excretion groups were distinguished suggesting the presence of long excretion markers, which are of remarkable interest in anti-doping analyses. The correspondence of some significant features with metabolites reported in the literature confirmed the rationale and usefulness of the proposed metabolomic approach. The presented study proposes a metabolomics workflow for the early detection and characterization of drug metabolites by untargeted urinary analysis to reduce the range of substances still excluded from routine screening. Its application has detected minor steroid metabolites, as well as unexpected endogenous alterations, proving to be an alternative strategy that can allow gathering a more complete range of information in the antidoping field.

The macrophages regulate intestinal motility dysfunction through the PGE2 Ptger3 axis during Klebsiella pneumonia sepsis.

Gut motility dysfunction, the most common complication of post-septic organ dysfunction, depends on immune and neuronal cells. This study aimed to investigate the mechanisms that activate these cells and the contribution of macrophages to the recovery of intestinal motility dysfunction after sepsis. Postoperative gut motility dysfunction was induced by establishing Klebsiella pneumonia sepsis in mice with selective deletion of neutrophils and macrophages in the gut. The distribution of orally administered fluorescein isothiocyanate-dextran and carmine excretion time was used to determine the severity of small bowel disease. The effect of macrophages on intestinal motility was evaluated after prostaglandin E2 therapy. We found that muscular neutrophil infiltration leading to neuronal loss in the intestine muscle triggered intestinal motility dysfunction after pneumonia sepsis; however, reduced neutrophil infiltration did not improve intestinal motility dysfunction. Moreover, macrophage depletion aggravated gut motility dysfunction. The addition of macrophages directly to a smooth muscle was responsible for the recovery of intestinal motility. Our results suggest that a direct interaction between macrophages and smooth muscle is neurologically independent of the restoration of intestinal dysmotility.

Different Animal Metabolism Markers for Artemia Nauplii in Crude Protein Digestibility Assay for Lophiosilurus alexandri Larvae

This work aimed to develop a methodology for marking Artemia with different markers and evaluating their passage rate and digestibility in Lophiosilurus alexandri larvae of different ages. In the first phase, Artemia nauplii were marked 28 h after hatching with chromium oxide, titanium dioxide and NANOLIPE®. Contact times with the markers were 30, 60, 90 min at a concentration of 500 mg/L. Titanium dioxide had a higher concentration in Artemia than the other markers. The passage rate did not differ significantly between titanium and NANOLIPE®. In the second phase, the apparent digestibility of the protein in Artemia nauplii was evaluated for L. alexandri larvae at three ages. Average real protein digestibility at the three ages was 95.08%, demonstrating Artemia as a good quality food. All ages exhibited the same final time of fecal excretion of 30 min after feeding. Artemia nauplii presented satisfactory results for assimilation for all markers tested. In determining the digestibility of Artemia protein, we can conclude that chromium oxide and NANOLIPE® were good markers of animal metabolism and titanium dioxide was the worst marker of Artemia digestibility for the initial stages of L. alexandri.

Urinary iodine excretion and optimal time point for sampling when estimating 24-h urinary iodine.

Iodine deficiency may cause thyroid dysfunction. The iodine intake in a population is measured by urinary iodine concentration (UIC) in spot samples or 24-h urinary iodine excretion (24UIE). 24UIE is considered the gold standard and may be estimated using an equation including UIC, urinary creatinine concentration, sex and age (e24UIE). The aims of this study were to evaluate the preferable timing of UIC when using this equation and assess the variability of UIE. Sixty healthy non-smoking women (n 31) and men (n 29) were included in Gothenburg, Sweden. Twelve urine samples were collected at six fixed times on two separate days. Variability was calculated for UIC, 24UIE, e24UIE, iodine excretion per hour (iHr) and UIC adjusted for creatinine and specific gravity. Median 24UIE was 156 µg/24 h and the median UIC (all spot samples) was 104 µg/l. UIC (P < 0·001), 24UIE (P = 0·001) and e24UIE (P < 0·001) were significantly higher in men. e24UIE was relatively similar to 24UIE. However, when e24UIE was calculated from UIC in the first void, it was about 15 % lower than 24UIE (P < 0·001). iHr was lowest in the morning and highest in the afternoon. Median iHr was higher in men (7·4 v. 5·3 µg/h, P < 0·001). The variability of UIE was higher within individuals than between individuals. This study suggests that most time points for estimation of individual 24UIE are appropriate, but they should preferably not be collected in the first void.

Effect of electroacupuncture at "Tianshu" (ST25) on the regulation of nitrergic neurons in different intestinal segments in streptozotocin induced type 2 diabetic rats

To observe the effect of electroacupuncture (EA) at "Tianshu" (ST25) on nitrergic neurons in jejunum and distal colon in type 2 diabetic rats, so as to explore its mechanism of regulating different intestinal segments. Twenty-four SD rats were randomly divided into control, model and EA groups (n=8 in each group). The diabetes model was established by intraperitoneal injection of streptozotocin (35 mg/kg) and high-sugar and high-fat diet for 2 weeks. EA (2 Hz/15 Hz, 2 mA) was applied to bilateral ST25 for 20 min, once a day, 6 days a week for 4 weeks. The intestinal motility was evaluated by observing the first red stool excretion time and the distal colon bead excretion time. HE staining was used to observe the histological changes of jejunum and distal colon. The positive expression and protein expression of intestinal total neuronal marker protein gene product 9.5（PGP9.5） and neuronal nitric oxide synthase (nNOS) in jejunum and distal colon were detected by immunofluorescence staining and Western blot, respectively. After modeling, the blood glucose was significantly increased (P<0.01), the first red stool excretion time and the distal colon bead excretion time were shortened (P<0.01), the expression levels of PGP9.5 and nNOS in jejunum and distal colon were decreased (P<0.01) in the model group relevant to the control group. After treatment, compared with the model group, the blood glucose was decreased (P<0.01), the first red stool excretion time and the distal colon bead excretion time were prolonged (P<0.01, P<0.05), and the expression levels of PGP9.5 and nNOS in jejunum and distal colon were increased (P<0.05, P<0.01) in the EA group. HE staining showed disordered structure in intestinal mucosa of the jejunum and distal colon, and reduction of the number of goblet cells in the model group, which was relatively milder in the EA group. EA can effectively improve the intestinal mucosal damage and restore intestinal motor function in type 2 diabetic rats, which may be related to its function in regulating the number of nitrergic neurons in the intestinal nervous system.

Magnetic resonance urography: a practical approach to preparation, protocol and interpretation.

Magnetic resonance urography (MRU) is an important MRI application that provides noninvasive comprehensive morphological and functional evaluation of the kidneys and urinary tract. It can be used to assess congenital anomalies of the kidney and urinary tract, which often present as urinary tract dilation. In children, MRU allows for high tissue contrast and high spatial resolution without requiring ionizing radiation. Magnetic resonance urography requires patient preparation in the form of pre-examination intravenous hydration, placement of a urinary catheter, and the administration of diuretics at the time of the exam. The imaging protocol is based on T2-weighted images for anatomical assessment and dynamic post-contrast images for functional evaluation. These images are then used to generate quantitative and graphic results including contrast transit and excretion time as well as to calculate differential renal function. This review focuses on a simple approach to pediatric MRU acquisition and interpretation based on clinical cases and the authors' experience.

Metabolic study of methoxpropamine (MXPr) in mice and determination of its metabolites in biological samples by UHPLC-QTOF-HRMS

The main metabolites of methoxpropamine (MXPr) and the evaluation of their formation and excretion over time, using biological samples from mice, was investigated. MXPr is an arylcyclohexylamine dissociative drug structurally similar to 3-MeO-PCE, ketamine and deschloroketamine, which was recently identified on the European illegal market and added to the class of New Psychoactive Substances (NPS). Forensic laboratories are often required to identify new drugs and their metabolites, albeit the latter are yet not known or their reference standards are lacking. Therefore, we performed the investigation of methoxpropamine (MXPr) metabolism, in order to elucidate the distribution of the parent drug and its metabolites in urine and plasma over time. Furthermore, body hair was collected and analysed. Initially, urine samples from 16 mice were collected every hour for 6 consecutive hours. Furthermore, one sample was collected after 12 and 24 hours from the administration of increasing doses of MXPr (1-3-10 mg/kg). After 16 days of wash out, two doses of 1 and 3 mg/kg were administered and plasma samples were collected. After adding ketamine-d4, urine and plasma were diluted 1:3 with methanol/acetonitrile (95:5) and 5 μL were injected into the UHPLC-QTOF-HRMS. Based on the typical phase I and II metabolic reaction, the metabolic pattern was hypothesized. The tentative metabolites were identified by means of the fragmentation patterns, the exact masses of both their precursor and fragment ions. Finally, body hair collected one month before and after the injection of MXPr was analyzed following a specific procedure for keratin matrix. A wide array of metabolites was identified in urine, including normethoxpropamine, desmethylmethoxpropamine, dihydromethoxpropamine, desmethyl-normethoxpropamine, dihydro-desmethylmethoxpropamine and phase II glucuronide conjugates. Furthermore, the metabolic pathway that produces desmethylmethoxpropamine-glucoronide was identified as the preferential pathway for the elimination of MXPr from the body. In fact, 1 h after the administration, an abundant peak of excretion was observed with an average intensity 16 times higher than the free phase desmetyl-MXPr. The presence of normethoxpropamine (NorMXPr) in urine and plasma is also relevant, following the typical metabolic pathways based on dealkylation. Since norketamine is known to be pharmacologically active, so it may be for NorMXPr, thus probably contributing to the toxicity of this substance. In addition, desmethylmethoxpropamine-glucoronide, together with other minor products of metabolism, were detectable in urine up to 24 h after injection. Finally, MXPr and the major phase I metabolites were observed also in plasma (average concentration for the parent drug: 1.0 ng/mL) and hair (average concentration for the parent drug: 0.18 ng/mg). Few phases I and II metabolites were confirmed to be present in body hair, approximately one month after body administration. Some interesting differences were observed in the metabolites generated over time between female and male mice, a phenomenon that could be related to sexually dimorphic metabolism, both involving phase I and phase II enzymes. A comprehensive workflow for the detection of MXPr and its I and II phase metabolites are presented. Data acquisition included a TOF-MS high-resolution scan combined with TOF-MS/MS acquisition and demonstrated a considerable capability to detect and identify target compounds with high resolution levels. Thanks to this study, the identification of MXPr metabolites and their excretion times from the body was possible. The introduction of new target compounds will improve the efficiency of toxicological screening analysis on real samples and will extend the window of detection of the dissociative drug MXPr in biological matrices. In the future, human urine samples will be analyzed to confirm the detectability of the tentative metabolites.