Soluble fluoride in Na2FPO3/CaCO3-based toothpaste as an indicator of systemically bioavailable fluoride

Fluoride chemically soluble in toothpaste is an indicator of fluoride bioavailability when the teeth are brushed and the same should be expected systemically when toothpaste is ingested. A four-phases study was conducted, in which eight participants were subjected in each phase to one of the assigned treatment groups: Group I: fresh sample of a Na2FPO3/CaCO3 toothpaste with 1,334 μg F/g of total soluble fluoride (TSF); groups II–IV: aged samples of toothpaste presenting TSF concentrations of 1,128, 808, and 687 μg F/g, respectively. In all phases, the participants ingested an amount of toothpaste equivalent to 70.0 µg F/kg body weight, as total fluoride (TF). Blood was collected before (baseline) and up to 180 min after toothpaste ingestion as indicator of fluoride bioavailability. Total urine (24 h before and 24 h after ingestion) was collected as indicator of absorbed fluoride that was excreted. F concentration in blood plasma and urine was determined with a fluoride ion-specific electrode. The areas under the curve of F concentration vs. time (AUC=ng F/ml x min) and the peaks of fluoride concentration in blood plasma (Cmax) were calculated. The net amount of fluoride excreted (mg/day) was calculated by subtraction. A significant correlation of the amount (mg) of TSF ingested was found between the AUC (r= 0.76; p<0.01) and Cmax (r= 0.86; p<0.01) in plasma, and the fluoride excreted (r= 0.65; p<0.01). For TF no statistical correlations were found (p>0.05). Data suggest that the concentration of TSF found in Na2FPO3/CaCO3-based toothpastes is a useful predictor of how much fluoride will be systemically bioavailable when this type of formulation is ingested.

PSI-2 A procedure for urine collection in pre-weaned heifer calves

Urine collection is a useful tool to analyze intestinal permeability in cattle for research and diagnostic purposes. However, urine sampling techniques often rely on total waste collection, which reduces the ability to perform more frequent sampling and obtain accurate and sterile urine volumes. A potential alternative is urethral catheters, which have been used in cows and weaned heifers. However, urethral catheters have not been thoroughly tested in pre-weaned dairy heifer calves. The study objective was to develop a urethral catheter placement procedure in pre-weaned heifer calves for continuous and accurate urine collection. Fifteen Holstein heifer calves had catheters placed at 8 ± 2 days (37.5 ± 3.38 kg BW) and 40 ± 2 days (59.3 ± 5.38 kg BW) of age. During the procedure, calves were individually housed (1.87 m2/calf) and restrained. The vulva was cleaned using betadine and 70% ethanol and then a sterile, lubricated 8.9 cm speculum was inserted into the vagina. A sterile guidewire (145 cm x 0.89 cm) was inserted into a lubricated sterile 10 FR catheter. The catheter was inserted into the urethral opening (~5–7 cm into vagina), guided into the bladder, and the catheter balloon was filled with water (10 mL). The guide wire was removed, and urine flow confirmed correct placement before a 4 L urinary drainage bag was attached to the catheter. Individual calf health observations were made twice over a 24-hour period and included vaginal discharge, bleeding, redness or inflammation, and tissue discharge in the urine. Occurrence rate was determined using PROC FREQ in SAS 9.4. Regardless of catheterization timing, bleeding and tissue discharge occurred at a rate of 3.33% ± 0.18, and vaginal discharge and inflammation occurred at a rate of 6.70% ± 0.25. In summary, this procedure is a viable method for total urine collection in pre-weaning heifer calves.

337 Efficient RFI Bulls Presented Lower Fractional Synthesis Rates of Plasma Proteins When Fed Corn but Not Grass Based Diets

Protein turnover (PT), the continual synthesis and degradation of body proteins not leading to protein gain, is an essential high energy-demanding process. We assumed that PT might explain the between-animal variations of residual feed intake (RFI). The objective was to measure PT in extreme RFI cattle fed two contrasted diets (grass or corn-based). We conducted a RFI test for 84 days with 100 Charolais bulls and we selected the 32 most extreme (8 per diet and RFI group) for PT measurements using 1) the urinary 3-methyl-histidine to creatinine ratio, as a biomarker of the fractional protein degradation rate (FDR) of skeletal-muscle and 2) the isotopic N turnover rate measured in urine and plasma, as a proxy, respectively, of the whole-body FDR and the fractional protein synthesis rates (FSR) of plasma proteins. The 3-methyl-histidine and creatinine were determined from 10 d total urine collection. Isotopic N turnover in urine and plasma was evaluated by modelling the 15N depletion rate over 112 d following an isotopic N dietary change. Higher plasma FSR and higher skeletal-muscle and whole-body FDR were observed with corn-vs-grass diets (≥11%; P ≤ 0.03), in line with higher metabolizable protein and net energy intakes (≥10%, P = 0.001). Differences between extreme RFI animals were noted with the corn diets only, where efficient animals presented significant lower plasma FSR (-10%; P = 0.04) and numerically lower skeletal-muscle and whole-body FDR (-13% and - 8.9%; P &gt; 0.16 respectively) than non-efficient. Non-significant differences were probably due to an insufficient size of our experimental setup. Plasma FSR is related to the PS of hepatic exportation, hence the lower plasma FSR observed in efficient RFI animals fed corn diets may reflect a lower organs to carcass ratio. Altogether results suggests that efficient RFI bulls fed corn diets had a lower hepatic PT with no-significant changes of whole-body and skeletal muscle PT.

The Clinical Application of Urine Soluble CD163 in ANCA-Associated Vasculitis

Background Up to 70% of patients with ANCA-associated vasculitis (AAV) develop glomerulonephritis, with 26% progressing to ESKD. Diagnostic-grade and noninvasive tools to detect active renal inflammation are needed. Urinary soluble CD163 (sCD163) is a promising biomarker of active renal vasculitis, but a diagnostic-grade assay, assessment of its utility in prospective diagnosis of renal vasculitis flares, and evaluation of its utility in proteinuric states are needed. Methods We assessed a diagnostic-grade urinary sCD163 assay in (1) a real-world cohort of 405 patients with AAV and 121 healthy and 488 non-AAV disease controls; (2) a prospective multicenter study of 84 patients with potential renal vasculitis flare; (3) a longitudinal multicenter cohort of 65 patients with podocytopathy; and (4) a cohort of 29 patients with AAV (with or without proteinuria) and 10 controls. Results We established a diagnostic reference range, with a cutoff of 250 ng/mmol for active renal vasculitis (area under the curve [AUC], 0.978). Using this cutoff, urinary sCD163 was elevated in renal vasculitis flare (AUC, 0.95) but remained low in flare mimics, such as nonvasculitic acute kidney injury. Urinary sCD163's specificity declined in AAV patients with nephrotic-range proteinuria and in primary podocytopathy, with 62% of nephrotic patients displaying a "positive" urinary sCD163. In AAV patients with significant proteinuria, urinary sCD163 normalization to total urine protein rather than creatinine provided the greatest clinical utility for diagnosing active renal vasculitis. Conclusions Urinary sCD163 is elevated in renal vasculitis flare and remains low in flare mimics. Nonspecific protein leakage in nephrotic syndrome elevates urinary sCD163 in the absence of glomerular macrophage infiltration, resulting in false-positive results; this can be corrected with urine protein normalization.

Plasma Metabolome Profiling for the Diagnosis of Catecholamine Producing Tumors

ContextPheochromocytomas and paragangliomas (PPGL) cause catecholamine excess leading to a characteristic clinical phenotype. Intra-individual changes at metabolome level have been described after surgical PPGL removal. The value of metabolomics for the diagnosis of PPGL has not been studied yet.ObjectiveEvaluation of quantitative metabolomics as a diagnostic tool for PPGL.DesignTargeted metabolomics by liquid chromatography-tandem mass spectrometry of plasma specimens and statistical modeling using ML-based feature selection approaches in a clinically well characterized cohort study.PatientsProspectively enrolled patients (n=36, 17 female) from the Prospective Monoamine-producing Tumor Study (PMT) with hormonally active PPGL and 36 matched controls in whom PPGL was rigorously excluded.ResultsAmong 188 measured metabolites, only without considering false discovery rate, 4 exhibited statistically significant differences between patients with PPGL and controls (histidine p=0.004, threonine p=0.008, lyso PC a C28:0 p=0.044, sum of hexoses p=0.018). Weak, but significant correlations for histidine, threonine and lyso PC a C28:0 with total urine catecholamine levels were identified. Only the sum of hexoses (reflecting glucose) showed significant correlations with plasma metanephrines.By using ML-based feature selection approaches, we identified diagnostic signatures which all exhibited low accuracy and sensitivity. The best predictive value (sensitivity 87.5%, accuracy 67.3%) was obtained by using Gradient Boosting Machine Modelling.ConclusionsThe diabetogenic effect of catecholamine excess dominates the plasma metabolome in PPGL patients. While curative surgery for PPGL led to normalization of catecholamine-induced alterations of metabolomics in individual patients, plasma metabolomics are not useful for diagnostic purposes, most likely due to inter-individual variability.

Antibacterial Residue Excretion via Urine as an Indicator for Therapeutical Treatment Choice and Farm Waste Treatment

Many of the infectious diseases that affect livestock have bacteria as etiological agents. Thus, therapy is based on antimicrobials that leave the animal’s tissues mainly via urine, reaching the environment through slurry and waste water. Once there, antimicrobial residues may lead to antibacterial resistance as well as toxicity for plants, animals, or humans. Hence, the objective was to describe the rate of antimicrobial excretion in urine in order to select the most appropriate molecule while reducing harmful effects. Thus, 62 pigs were treated with sulfamethoxypyridazine, oxytetracycline, and enrofloxacin. Urine was collected through the withdrawal period and analysed via LC-MS/MS. Oxytetracycline had the slowest rate of degradation (a half-life time of 4.18 days) and the most extended elimination period in urine (over 2 months), followed by enrofloxacin (a half-life time of 1.48 days, total urine elimination in ca. 3 weeks) and sulfamethoxypyridazine (a half-life time of 0.49 days, total urine elimination in ca. 1 week). Bacterial sensitivity and recommendations for responsible use are limiting when selecting the treatment. Nevertheless, with similar effectiveness, sulfamethoxypyridazine would be the choice, as waste treatment would only need to be implemented for 1 week after treatment. Thus, more in-depth knowledge regarding antibacterial elimination would improve resource management, while protecting animals and consumers’ health.

Paraganglioma

Introduction: Paragangliomas are rare neuroendocrine tumors arising from extra-adrenal medullary neural crest derivatives. The terms pheochromocytoma and paraganglioma are often used interchangeably because morphologically and functionally these entities are almost the same. However, paragangliomas that arise in the adrenal medulla are called pheochromocytomas and those outside the adrenal gland are called paragangliomas. Paragangliomas are often discovered incidentally during imaging studies performed for other reasons. We report a case of a patient who had incidental finding of retroperitoneal paraganglioma on imaging done for evaluation of thigh and gluteal cellulitis. Clinical Case: A 24-year-old female presented with chief complaints of episodic headaches, multiple skin abscesses, fever and malaise. She has a history of Type 1DM and had been non-compliant on insulin. Vitals on presentation was significant for tachycardia with heart rate of 124, blood pressure of 119/81, respiratory rate of 16. Initial labs were consistent with DKA. CT abdomen and pelvis done for further evaluation of gluteal and thigh abscess showed incidental finding of a 3.4 cm retroperitoneal/para-aortic well-circumscribed rim-enhancing mass with central hypoenhancing component, suspicious for neoplasm such as paraganglioma. Biochemical testing was performed. 24-hour urine catecholamine levels obtained showed elevated urine norepinephrine level of 1008μg/day (reference range 15-100μg/day), urine dopamine 410μg/day (reference range 65- 400μg/day), urine epinephrine less than 2μg/day (reference range 0-20μg/day). Total urine catecholamines was elevated 1008μg/day (reference range 15-100μg/day). These results confirmed diagnosis of paraganglioma. Treatment options were discussed with the patient including surgery for removal of paraganglioma which she has currently declined. Conclusion: Diagnosis of a paraganglioma can usually be made using biochemical and radiographic testing. All patients with paraganglioma should be tested for hypersecretion of catecholamines in a 24-hour urine or serum collection, even if they do not present with a clinical picture of catecholamine hypersecretion. Importantly these extra-adrenal tumors do not have the enzymatic capacity to form epinephrine from norepinephrine as was exemplified by our case. For catecholamine-secreting tumors, biochemical diagnosis should be followed by radiological evaluation (typically either CT or MRI of the abdomen and pelvis) to locate the tumor. Treatment options are dependent on location of tumor, size, presence of symptoms and if there is metastatic disease present.

Clinical case of gestational diabetes insipidus

Diabetes insipidus is a rare endocrinological disease and occurs in 2–4 per 100,000 pregnancies. Diagnosis of gestational diabetes insipidus is very difficult because it develops against the background of physio­logical mechanisms that accompany pregnancy: thirst threshold decreases leading to polydipsia and plasma osmolarity decreases causing hypotonic polyuria. Understanding of pathophysiology of the disorder is very important for further management of these vulnerable patients. A 32-year-old patient at 36 weeks of gestation, primigravida, was referred to an endocrinologist with complaints of polyuria (6.5 l/day), nocturia — up to 5 times, severe polydipsia. At 12 weeks of gestation, there was a risk of abortion for prevention of which the patient received progesterone 100 mg intravaginally twice a day until 34 weeks. She has a history of subacute thyroiditis, with no family history of endocrine pathology. Physical examination revealed a decrease in skin turgor, blood pressure 110/85 mm Hg. Heart rate 115 bpm, weight 71 kg (body mass index 26.9 kg/m2). The patient was at high risk of developing preeclampsia. Laboratory data: analysis of urine according to Zimnitsky: volume per day — 6.8 l, specific gravity in portions: 1.012; 1.008; 1.010; 1.005; 1.012; 1.014; 1.010. Total blood count, total urine test, serum sodium and potassium, liver function tests, level of thyroid-stimulating hormone, free thyroxine, thyroid peroxidase antibodies and morning free cortisol level were normal. The patient was administered desmopressin 10 μg intranasally twice daily. Six weeks after delivery, desmopressin was stopped and she had no further evidence of polyuria, polydipsia or nocturia.

Ameliorative effect of apelin-13 against renal complications in L-NAME-induced preeclampsia in rats

Pre-eclampsia (PE) accompanying acute liver and kidney injury has remained a master cause of both fetal and maternal mortality and morbidity. Vasoactive mediators, oxidative stress and inflammatory imbalanceshave an important role in PE pathogenesis. Apelin is an adipokine that improves endothelial dysfunction; has anti-inflammatory and antioxidant effects; moreover, its level reduced during PE. This study aimed to explore the effects of apelin-13 administration on preeclampsia-associated renal dysfunction and proteinuria. Thirty-three pregnant female rats were divided into three groups; group: 1 (normal pregnant rats), group: 2 (preeclamptic rats); where rats were injected subcutaneously with 75 mg L-NAME/ kg body weight/day beginning from 9th to 20th day of pregnancy andgroup 3 (apelin-13 treated preeclamptic rats); In which L-NAME-induced preeclamptic rats were subcutaneously injected with 6 × 10−8 mol apelin-13/kg body weight/twice daily starting from 6th to 20th day of pregnancy. In all groups, mean arterial blood pressure, total urine protein, serum urea, creatinine, nitric oxide (NO), endothelin-1 (ET-1), interleukin–6 (IL-6) and malondialdhyde (MDA) were measured. Histopathological examination of kidney tissues was also done. preeclamptic rats showed significantly increased mean arterial blood pressure, total urine proteins, serum urea, creatinine, ET-1, IL-6, and MDA, but revealed a significantly decreased serum NO level. On the other hand, apelin treatment significantly improved these parameters together with amelioration of kidney histoarchitecture in the treated group. In conclusion, apelin may be a potentially curative candidate for prohibiting kidney damage and have a therapeutic benefit in PE rat models.