Dunnett's Multiple Comparison Test Research Articles

Sales trends of psychotropic drugs in the COVID-19 pandemic: A national database study in Brazil.

Background: The social restrictions among coronavirus disease 2019 (COVID-19) pandemic have posed a thoughtful risk to mental health and have implications in the use of drugs, including antidepressants, anxiolytics and other psychotropics. Objective: This study analyzed the sales data of the psychotropics prescribed in Brazil, in order to verify the change in consumption trends of these drugs during the COVID-19 pandemic. Methods: This interrupted time-series analyzed psychotropic sales data, between January 2014 and July 2021, using the National System of Controlled Products Management from The Brazilian Health Regulatory Agency. The monthly mean DDDs per 1,000 inhabitants per day of psychotropic drugs was evaluated by analysis of variance (ANOVA) followed by Dunnett Multiple Comparisons Test. The changes in monthly trends in the use of the psychotropic studied were evaluated by Joinpoint regression. Results: During the period studied, clonazepam, alprazolam, zolpidem and escitalopram were the most sold psychotropic drugs in Brazil. According to Joinpoint regression, an upward trend was observed in sales during the pandemic of pregabalin, escitalopram, lithium, desvenlafaxine, citalopram, buproprion and amitriptyline. An increase in psychotropic consumption was noted throughout the pandemic period, with the maximum consumption (2.61 DDDs) occurring in April 2021, with a downward trend in consumption that accompanied the drop in the number of deaths. Conclusions: The increase in sales, mainly of antidepressants during the COVID-19 pandemic, draws attention to issues related to the mental health of the Brazilian population and on the need for greater monitoring in the dispensing of these drugs.

Functional and morphological study of the effects of carvacrol on smooth muscle of the thoracic aorta in the rat.

The leaves of Origanum are widely used in herbal medicine hence of having many beneficial ingredients, one of these important compounds is Carvacrol. The inhibitory effect of Carvacrol was the core of this study by applying different kinds of stimulants to smooth muscles in the wall of thoracic aorta in rats. To investigate the pharmacological effects of Carvacrol, the main active ingredient present in the medicinal plant Origanum, on the contractile activity and morphology of the smooth muscle of the rat thoracic aorta. After the thoracic aorta arteries were isolated and prepared for the experiments, each thoracic aorta was cut into 5-mm ring segments; different stimulants were used (Potassium Chloride, Norepinephrine, U46619, and α,β-methylene ATP) in the presence and absence of Carvacrol on four groups of rats. The isolated rings were placed and connected to a force transducer which in turn linked to a data acquisition system via an amplifier to record the effect of each stimulant. GraphPad Prism version 5.02 for Windows, one-way analysis of variance followed by Dunnett's multiple comparison test. It was found out that Carvacrol obstructs the contractile responses elicited by exogenous NA, KCl, U46619, and α,β-methylene ATP in a concentration dependent manner. The addition of Carvacrol in the experimental rats showed an increase in the thickness of tunica media as evident by the number of smooth muscle layers and laminae of elastic fibers. It was found that Carvacrol reduced the vascular smooth muscle contractility in the rat thoracic aorta. The mechanism of action is presumed to be achieved through interfering with the mobilization of both intracellular and extracellular Ca2+ through different receptors. Furthermore, it might be suggested that Carvacrol in high doses stimulates smooth muscles in the wall of aorta leading to an increase in the thickness of tunica media layer.

The incidence and characterization of weight gain associated with MEK inhibitors in pediatric patients

Mitogen-activated protein kinase enzyme (MEK) inhibitors are used in the treatment of pediatric patients with neurofibromatosis, low grade glioma, and astrocytoma, and may demonstrate a unique side effect profile in this population. Inhibition of MEK has been shown to decrease interleukin (IL)-6 production, a proinflammatory cytokine. The inhibition of IL-6 and other proinflammatory cytokines is thought to decrease muscle wasting and may contribute to weight gain. However, there is limited information on the association of MEK inhibition and weight gain in children and adolescents. This study aimed to characterize and define the incidence of significant weight gain associated with MEK inhibitors in pediatric patients. This was a retrospective chart review conducted at a tertiary pediatric hospital. Children 1-18.99 years old were included if they started a MEK inhibitor from July 1, 2013-October 31, 2021, and continued therapy for at least 6 months. Significant weight gain was defined as ≥5% increase in patient's weight-for-age percentile. Sixty-seven patients were included in the analysis. Sixty-two received trametinib and 5 received selumetinib. An increase in weight-for-age percentile ≥5% was seen in 60% of patients receiving selumetinib and 56% on trametinib. The Dunnett's multiple comparisons test revealed a difference in weight-for-age percentile from baseline to end of data collection (p=.0173). Patients who were obese at baseline were more likely to lose weight during treatment, while underweight patients increased in weight-for-age percentiles. Weight gain may be a notable side effect associated with the use of MEK inhibitors in pediatric patients.

Downregulation of MICA/B tumor surface expressions and augmented soluble MICA serum levels correlate with disease stage in breast cancer.

In this study, the profiling of the expression of major histocompatibility complex (MHC) class I-related chain A and B (MICA/B) in human breast cancer tumor tissue, saliva, and urine samples of breast cancer patients and control is carried out. MICA/B is ligand of NKG2D receptor expressed on malignant cells. The release of MICA/B from tumor tissue comprises an immune escape mechanism that impairs antitumor immunity. Based on this literature we explored the potential of soluble MICA (sMICA) as a marker in breast cancer (BC). The expression was profiled by using immunohistochemistry (MICA/B), western blot (MICA/B) and ELISA (MICA). The optical density of western blot of MICA/B in different stages of BC illustrated significant difference as per one way analysis of variance and significant difference with stage III and IV by Dunnett's multiple comparisons test respectively. Analysis of sMICA in serum, saliva and urine of BC patients revealed significantly higher levels (median 41.0 ± 4.1 pg/ml in pre-treatment sera, 181.9 ± 1.6 pg/ml in saliva and 90.7 ± 1.7 pg/ml in urine) than in control (median <1.2 pg/ml). The elevated levels of sMICA were related to the cancer stage. The elevated levels of sMICA were observed in patients with well differentiated cancer while the poor expression of sMICA was observed in patients with poorly differentiated tumors. Tumor immunity is impaired by the release of MICA in the biofluids and may be useful for detection and diagnosis of the stage of BC.

Mesoporous CuS/SiO2 as a sulfamethoxazole loading carrier against Escherichia coli and Staphylococcus aureus

Metallic nanoparticles (NPs) (Ag, Cu, CuO and CuS) have antibacterial properties that are closely related to the size and morphology that enable a large surface-to-volume ratio. In solution, metallic NPs are likely to be agglomerated or precipitated, thus decreasing their biocidal activity. The surface functionalization of copper sulfide NPs on mesoporous SiO2 spheres (PMSi) was successfully achieved and characterized by XRD, SEM, TEM, UV–vis and nitrogen physisorption. Since the incidence of allergies in humans caused by sulfamethoxazole (SMX), a water insoluble bacteriostatic antibiotic (commonly used in combination with trimethoprim), has been detected to be increasing, in this research work, it was decided to study the antibacterial activity of PMSi loaded with SMX against Escherichia coli and Staphylococcus aureus, Gram-negative and Gram-positive bacteria, respectively. The antibacterial synergistic performance displayed by loading SMX on CuSNPs was studied by the minimum inhibitory concentration (MIC) method. The ANOVA analysis with Dunnett's multiple comparison tests showed a statistically significant difference between the CONTROL and PMSi composites unloaded and loaded with SMX on E. coli and S. aureus. CuS and Cu functionalized on PMSi exhibited a good spectrum of antimicrobial activity against Gram-positive (S. aureus) and Gram-negative (E. coli) bacteria, in contrast with the current literature.

To Determine Therapeutic Effect of Curcuma Gel on Gingivitis as an Adjunct to Scaling

Context: Gingivitis is among the most prevalent periodontal disease occurring in the oral cavity, which is plaque induced and if untreated, can progress to periodontitis. Various adjunct chemotherapeutic agents are available for plaque control with their own pros and cons. Hence, a plaque-controlling therapeutic agent which is readily available at low cost and without any side effects is the need of the hour. According to the ancient texts, Ayurveda and the traditional culture of India, Curcuma longa is used externally for inflammation of skin and mucosa. Aims: To evaluate the therapeutic effect of curcuma gel on gingival health and its possible usage in the treatment of gingivitis. Settings and Design: This study comprises hundred participants with gingivitis which were divided randomly in two groups. Subjects and Methods: Experimental group – Scaling and topical application of curcuma gel and control group – Scaling alone without the topical application of curcuma gel. Clinical parameters (Gingival Index, Gingival Bleeding Index, and Plaque Index) were assessed at different time intervals (0, 7, 14, and 21 days) in both experimental and control groups. Statistical Analysis Used: Within groups, a repeated measures ANOVA model was used for repeated observations over a period which is followed by the Dunnett test for multiple comparisons. Experimental and control groups were compared for normally distributed data by Student's t-test. Whereas for skewed data, the Wilcoxon test or Mann–Whitney U-test was employed. Results: Statistically significant improvement of gingival health parameters was seen in the group which received topical curcuma oral gel application after scaling compared with the control group. Conclusions: Significant improvement in clinical parameters of gingivitis on the application of Curcuma oral gel after scaling with no obvious adverse effects suggesting its possible use as a topical adjunct treatment in clinical or community level prevention as well as treatment programs in the future.

Whole-genome sequencing of biofilm-forming and chromium-resistant mangrove fungus Aspergillus niger BSC-1

Filamentous fungus Aspergillus niger has gained significant industrial and ecological value due to its great potential in enzymatic activities. The present study reports the complete genome sequence of A. niger BSC-1 which was isolated from Indian Sundarban mangrove ecosystem. The study revealed that the genome of A. niger BSC-1 was 35.1 Mbp assembled in 40 scaffolds with 49.2% GC content. A total of 10,709 genes were reported out of which 10,535 genes were predicted for encoding the proteins. BUSCO assessment showed 98.6% of genome completeness indicating high quality genome sequencing. The genome sequencing of A. niger BSC-1 revealed the presence of rodA and exgA genes for initial adhesion to surface and Ags genes for matrix formation, during biofilm growth. OrthoVenn2 analysis revealed that A.niger BSC-1 shared 9552 gene clusters with the reference strain A. niger CBS554.65. Semi-quantitative RT-PCR analysis unveiled the role of Ags1 and P-type ATPase in fungal biofilm formation and chromium (Cr) resistance, respectively. During biofilm growth the expression of Ags1 significantly (P < 0.0001; two-way ANOVA followed by Sidak's multiple comparisons test) increased with respect to planktonic culture revealing the possible involvement of Ags1 in biofilm matrix formation. Expression of P-type ATPase gene was significantly upregulated (P < 0.0001; one-way ANOVA followed by Dunnett's multiple comparisons test) with the increasing chromium concentration in the fungal culture. Besides, several other genes encoding metalloprotease, copper and zinc binding proteins, and NADH-dependent oxidoreductase were also found in the genome of A. niger BSC-1. These proteins are also involved in heavy metal tolerance and nanofabrication indicating that this filamentous fungus A. niger BSC-1 could be potentially utilized for chromium detoxification through biofilm or nanobiremediation.

Deep anterior lamellar keratoplasty in keratoconus

Aims: To analyse the outcome of DALK in patients with keratoconus in terms of best corrected and uncorrected visual acuity, average K and topographic astigmatism. Settings and design: Prospective interventional case series study Material and methods: 28 eyes of 28 patients diagnosed with keratoconus were evaluated from August 2009 – July 2011 in a tertiary eye care hospital. All patients underwent DALK and outcomes were assessed on 4 parameters: Average Keratometric reading, topographic astigmatism, UCVA and BCVA alongwith epidemiological analysis. Statistical analysis used: ANOVA testing and Dunnett multiple comparison test. Results: We had evaluated 28 eyes of 28patients (10 males and 18 females) in the age group of 15 to 35 years. Cases were classified as moderate (4) and severe (24) category. There was a statistical significant decrease Average K and Topographic Astigmatism in post op period which was evaluated at 1, 3 and 6 months. (p value &lt;0.05). From the sample size, allof the patients showed improvement in visual acuity by more than one Snellens. No significant post operative complications occurred. Conclusion: DALK is a safe and effective treatment for severe keratoconus, corneal scarring and contact lens intolerance.

Patient Reported Outcomes and Financial Toxicity in Head and Neck Cancer (PaRTNer): Longitudinal Assessment of Financial Toxicity and an Educational Intervention from a Pilot Study

<h3>Purpose/Objective(s)</h3> Patients undergoing definitive head and neck cancer (HNC) treatment are at risk for high out-of-pocket (OOP) costs given complex treatment and high symptom burden. The PaRTNer pilot study was designed to assess costs, financial toxicity, and the benefit of education for patients undergoing definitive radiation (RT) for HNC. <h3>Materials/Methods</h3> Eligibility: adult patients with newly diagnosed non-metastatic HNC undergoing RT for definitive cancer treatment. Baseline survey collected demographics, cancer details, OOP costs, and COmprehensive Score for financial Toxicity (COST) score to measure financial toxicity. Patients received a binder with educational materials regarding financial toxicity and logs to document OOP costs. Repeat surveys at 3 and 6 months (mo) evaluated OOP costs, financial toxicity, and benefits of the provided binder. COST scores over time were evaluated by mixed-effects model (restricted maximum likelihood) and Dunnett's multiple comparisons test in scientific 2-D graphing and statistics software. <h3>Results</h3> 60 patients were enrolled from 2019-2021 with 58 (baseline), 52 (3 mo), and 48 (6 mo) completing surveys longitudinally. Most were white (76%), non-Hispanic (98%), and male (67%), with median age of 61 (range 42-86). Oropharynx was the most common primary disease site (48%), and 48% received concurrent chemotherapy with RT. 59% were working part-time or full-time, and 55% had employer-sponsored private insurance. Annual household income was >$60,000 for 55% of patients. Mean COST scores at baseline, 3 mo, and 6 mo post-RT were 21.3, 19.2, and 18.6, respectively (adjusted p=0.002, baseline vs 6 mo), with lower COST score indicating worse financial toxicity. Cumulative median OOP cost was $3460 (Range: $16-20614). At baseline, 64% had little or no understanding about the costs of their cancer care, and 69% felt that educational materials about costs would be helpful. 56% and 71% of patients at 3 and 6 mo post-RT felt that the binder of educational materials provided at study enrollment was very helpful. At 3 mo post-RT, 96% indicated this information should be given to all cancer patients. <h3>Conclusion</h3> Patients undergoing definitive RT for HNC experienced borderline financial toxicity prior to RT and a small but significant increase in financial toxicity at 6 mo post-RT. Most patients had limited baseline understanding about cancer costs and found study educational materials increasingly helpful over time. This pilot study demonstrates feasibility of longitudinal assessment of financial toxicity for HNC patients undergoing RT and highlights the potential utility of educational interventions, warranting assessment in future prospective studies. This was a single-institution, prospective longitudinal study (NCT3506451).

Efficacies of two medicinal plants [Goma Guar Walmart (Cyamopsis tetragonolobus L.) and Cinnamon (Cinnamomum verum)] versus efficacy of allopathic treatment [Sil–Norboral (Glibenclamide 5 mg/Metformin 1000 mg + Janumet (Sitagliptin 50 mg/Metformin 850 mg)

The objective of this study was to statistically compare –using the One–Way Analysis of Variance and Dunnett's multiple comparisons test– the efficacy of Cyamopsis tetragonolobus L. and Cinnamomum verum versus the efficacy of the allopathic treatment Sil–Norboral (Glibenclamide 5 mg/Metformin 1000 mg) + Janumet (Sitagliptin 50 mg/Metformin 850 mg) as hypoglycemic agents in patients with type 2 diabetes mellitus who attend the "Dr. Agustin O'Horán" General Hospital for medical care. The epistemological approach is quantitative, probabilistic or positivistic. The study design corresponds to that of a therapeutic experimental epidemiological study (therapeutic clinical trial) with prospective temporality. Forty–eight patients with type 2 diabetes mellitus were studied. The 48 patients were randomly assigned to three groups: two experimental groups and one control group. Each group was made up of 16 patients. The first experimental group was given Cyamopsis tetragonolobus L. (twelve dry leaves of Cyamopsis tetragonolobus L. and 500 ml of hot water); pour the 500 ml of hot water into a container and add the twelve dry leaves of Cyamopsis tetragonolobus L.; take 250 ml in the morning and 250 ml at night); the second experimental group was given Cinnamomum verum (two g of dry leaves of Cinnamomum verum and 500 ml of hot water; pour the 500 ml of hot water into a container and add two g of dry leaves of Cinnamomum verum; take 250 ml in the morning and 250 ml at night); and the control group was given the Sil–Norboral allopathic treatment (Glibenclamide 5 mg/Metformin 1000 mg) + Janumet (Sitagliptin 50 mg/Metformin 850 mg). In ascending numerical order, the arithmetic means of the hematic glucose values ​​corresponded to the following treatments: Cyamopsis tetragonolobus L. (84.06 mg/100mL); Cinnamomum verum (88.63mg/100mL); and Sil–Norboral (Glibenclamide 5 mg/Metformin 1000 mg) + Janumet (Sitagliptin 50 mg/Metformin 850 mg) (122.25 mg/100mL). The One–Way Analysis of Variance reported a value of F= 56.03 with a value of p= 0.0000, which indicates a statistically significant difference between a pair or between more than one pair of arithmetic means. Dunnett's multiple comparisons test showed statistically significant differences between the treatment with the medicinal plant Cyamopsis tetragonolobus L. and the allopathic treatment Sil–Norboral + Janumet: p= 0000. Likewise, Dunnett's multiple comparisons test showed a statistically significant difference between the treatment with the medicinal plant Cinnamomum verum and the allopathic treatment Sil–Norboral + Janumet: p= 0.0000. It is concluded that Cyamopsis tetragonolobus L. is, from a numerical and not a statistical point of view, the best hypoglycemic agent for the treatment of type 2 diabetes mellitus.

Effects of Cellgevity® on the milt quality of catfish,Clarias gariepinus extended in sodium citrate during chilled storage

Cellgevity® is a supplement reported to comprise mostly D-Ribose and L-Cysteine enriched glutathione, known to be an effective antioxidant that improves spermatozoa quality. However, its effect on milt characteristics has not been reported. This study, therefore, aimed to evaluate the effects of Cellgevity® on the milt quality of catfish (Clarias gariepinus) extended in sodium citrate during chilled storage. Pooled milt sample from three fishes was divided into three groups (T1, T2 and T3). The milt was extended in sodium citrate, and each group in triplicate was supplemented with Cellgevity® at 0 mg (T1), 125 mg (T2) and 250 mg (T3). The spermatozoa motility, concentration, viability and morphology were evaluated on days 0, 1, 2, 3, 4 and 5 of chilled storage. Data were expressed as mean ± standard deviation (SD) and analysed with a one-way analysis of variance (ANOVA) followed by Dunnett's multiple comparison test. Mean ± SD spermatozoa motility was significantly (P &lt; 0.001) lower in T2 and T3 than T1 before and during the first 3-days storage period. Mean ± (SD) spermatozoa concentration was significantly (P &lt; 0.001) higher in T2 and T3 than T1 before and throughout the 5-days storage period. Mean ± SD live spermatozoa were significantly (P &lt; 0.001) lower in T3 than T1 at day 2 of the storage. Mean ± SD total abnormal spermatozoa did not differ significantly (P &gt; 0.05) among the groups before and throughout the 5-days storage period. It was concluded that although supplementation of Cellgevity® at 125 mg and 250 mg in milt of catfish, extended in sodium citrate in chilled storage maintained the sperm cells alive and motile up to four days of the storage. However, it did not improve the milt quality. Hence, it should not be supplemented in sodium citrate extended milt of catfish, Clarias gariepinus in chilled storage.

Effect of heat-moisture treated brown rice crackers on postprandial flow-mediated dilation in adults with mild endothelial dysfunction

BackgroundEndothelial dysfunction is an early pathophysiological feature and independent predictor of a poor prognosis in most forms of cardiovascular disease. We evaluated the effect of brown rice crackers (BR-C) on endothelial function. MethodsEffect of heat-moisture treated (HMT) -BR-C on postprandial flow-mediated dilation (FMD) in adults with mild endothelial dysfunction was compared with that of BR-C and white rice crackers (WR-C) in 12 adults with mild endothelial dysfunction (less than 7.0% of FMD) by a randomized, single-blind, three-treatment three-period crossover trial (UMIN 000034898). Since we considered that the FMD increase was associated with the treatment of HMT-BR-C, we examined the effect of three possible factors: postprandial glucose levels, polyphenol content, and polyphenol release from the food matrix. ResultsMean pre-intake baseline FMD values of HMT-BR-C, BR-C, and WR-C were 4.9%, 5.1%, and 4.9%, respectively, and those values 1 h post-intake were 6.3%, 5.1%, and 4.8%, respectively. There was no difference in intergroup comparisons of FMD using Dunnett's multiple comparison test. There was a significant increase in FMD only in HMT-BR-C in intragroup comparisons (P = 0.042 by paired-t test). In comparison with BR-C, no significant difference was noted in the postprandial glucose level nor in the content of total polyphenols and ferulic acid derivatives in HMT-BR-C. However, the 70% ethanol extracted from HMT-BR-C contained a significantly larger amount of free and bound ferulic acids than from BR-C. ConclusionHMT-BR-C intake increased the postprandial FMD response.

New Cluster Analysis Method for Quantitative Dynamic Contrast-Enhanced MRI Assessing Tumor Heterogeneity Induced by a Tumor-Microenvironmental Ameliorator (E7130) Treatment to a Breast Cancer Mouse Model.

Dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) can provide insight into tumor perfusion. However, a method that can quantitatively measure the intra-tumor distribution of tumor voxel clusters with a distinct range of Ktrans and ve values remains insufficiently explored. Two-dimensional cluster analysis may quantify the distribution of a tumor voxel subregion with a distinct range of Ktrans and ve values in human breast cancer xenografts. Prospective longitudinal study. Twenty-two female athymic nude mice with MCF-7 xenograft, treated with E7130, a tumor-microenvironmental ameliorator, or saline. 9.4 Tesla, turbo rapid acquisition with relaxation enhancement, and spoiled gradient-echo sequences. We performed two-dimensional k-means clustering to identify tumor voxel clusters with a distinct range of Ktrans and ve values on Days 0, 2, and 5 after treatment, calculated the ratio of the number of tumor voxels in each cluster to the total number of tumor voxels, and measured the normalized distances defined as the ratio of the distance between each tumor voxel and the nearest tumor margin to a tumor radius. Unpaired t-tests, Dunnett's multiple comparison tests, and Chi-squared test were used. The largest and second largest clusters constituted 44.4% and 27.5% of all tumor voxels with cluster centroid values of Ktrans at 0.040 min-1 and 0.116 min-1 , and ve at 0.131 and 0.201, respectively. At baseline (Day 0), the average normalized distances for the largest and second largest clusters were 0.33 and 0.24, respectively. E7130-treated group showed the normalized distance of the initial largest cluster decreasing to 0.25, while that of the second largest cluster increasing to 0.31. Saline-treated group showed no change. A two-dimensional cluster analysis might quantify the spatial distribution of a tumor subregion with a distinct range of Ktrans and ve values. 1 TECHNICAL EFFICACY: Stage 1.

Comparison of non-invasive measurements of penile curvature in Peyronie's Disease

ABSTRACT Introduction Peyronie's disease (PD) is a fibrotic disorder of the tunica albuginea, leading to penile deformities. Non-invasive estimates of penile deformities are practical, but have low accuracy when compared to objective measurements with induced erection and a goniometer. Our aim was to evaluate the agreement between different non-invasive methods of measuring penile curvature and the objective measurement. Methods We retrospectively reviewed medical records of patients who underwent surgery in a sexual medicine tertiary center. During the clinical visit, pre-operatively, a set of different non-invasive estimates were performed including asking the patient to explain their primary curvature with a goniometer, a drawing of the penis and a malleable implant prosthesis to be molded. Also, at home photography were required with maximal erection. The agreement of those methods and intra-operative measurement using a goniometer after saline induced erection was analyzed. A tolerable error was considered if a difference smaller than 10o or 20% of the gold standard measurement was found. Results The sample consisted of 69 men with a mean age of 62 ± 6 y. The direction of the curvature was dorsal in 59%, left in 35%, right in 5% and ventral in 1%. Mean curvature with of estimates with the goniometer was 57o (±31), drawing 60 o (±28), Prosthesis 60o (±25), Photography 57 o (±27), and mean intra-operative measurement was 51 o (± 24). Overall precision considering the above mentioned tolerable error definition was 45%, and the single non-invasive method with better precision was Prosthesis with 46,5% fo concordance. There was no statistical difference between them (p=0.99) for concordance and disagreement by the Dunnett's multiple comparisons test. Conclusion In our comparison study of 4 non-invasive curvature assessment methods, we found a low precision (approximately 45%), regardless of the chosen method. Formal curvature assessment with induced erection and a goniometer appears to be imperative. Disclosure Work supported by industry: no.

Development of an Automated, High‐Throughput Assay to Detect Angiotensin AT2‐Receptor Agonistic Activity by Nitric Oxide Measurement In Vitro

BackgroundThe angiotensin AT2‐receptor (AT2R) is a key component within the protective arm of the renin‐angiotensin system inducing vasodilation by nitric oxide (NO) production. Currently, no high‐throughput assay is available to identify AT2R agonists in vitro, which could explain the low number of AT2R selective ligands in drug development programs.ObjectiveTo design and validate a high‐throughput method for detection of AT2R activation in vitro based on NO release.MethodsHuman aortic endothelial cells (HAEC) were seeded into 96‐well plates at a density of 5000 cells per well. 48 hours after seeding, cells were loaded for 30 minutes with the NO probe, DAF‐FM diacetate (5µM). Cells were stimulated for 15 minutes with the AT2R‐agonist C21 (1µM) or with classical NO‐releasing agents as positive controls: bradykinin (BK, 0.1µM), acetylcholine (ACh, 1µM) or Ang‐(1‐7) (MasR agonist, 0.1µM). Cells were fixed and fluorescence signals captured by fluorescence microscopy with automated, built‐in image analysis (ImageXpress Pico, Molecular Devices, USA). AT2R‐transfected (AT2R‐CHO) or non‐transfected (NT‐CHO) Chinese hamster ovary cells were cultured and loaded with DAF‐FM diacetate according to the same protocol as HAEC. Concentration‐response curves were obtained for C21, BK, Ang II and Ang‐(1‐5) (0.1nM to 10µM each) using the automated protocol for ImageXpress Pico described above on AT2R‐CHO. To evaluate whether effects of the compounds in AT2R‐CHO were AT2R specific, the response to these compounds at the highest concentration (10µM) was also tested in NT‐CHO.ResultsUsing primary HAEC we could show that the automated, high‐throughput workflow was able to detect NO release by AT2R agonists as efficiently as by classical NO‐releasing agents [C21: 16.16 ± 6.84%; Ang‐(1‐7): 25.45 ± 3.37%, BK: 19.30 ± 2.92%; ACh: 24.01 ± 4.24%; one‐way ANOVA followed by Dunnett’s multiple comparison test, p≤0.05 vs control, ≥3 independent experiments for all compounds). In AT2R‐CHO, we detected that both C21 and Ang II (AT2R agonists) increased NO release in a concentration‐dependent manner, while BK had no effect (AT2R‐CHO lacks BK receptors) at any tested concentration. Surprisingly Ang‐(1‐5), a degradation product of Ang II thought to be inactive, induced NO release in a concentration‐dependent manner in AT2R‐CHO. In NT‐CHO, none of the compounds stimulated NO release, indicating that the effects by C21, Ang II and Ang‐(1‐5) in AT2R‐CHO were AT2R specific.ConclusionMeasurement of NO release from AT2R‐CHO or HAEC by DAF‐FM diacetate in an automated way is suitable as high‐throughput assay for the identification of AT2R‐agonistic compounds in vitro. Application of the assay revealed that Ang‐(1‐5), which has been regarded as an inactive metabolite of Ang II, has AT2R agonistic properties.

Consumption Trends of Antibiotics in Brazil During the COVID-19 Pandemic.

Background: In 2019, a new type of coronavirus emerged and spread to the rest of the world. Numerous drugs were identified as possible treatments. Among the candidates for possible treatment was azithromycin alone or in combination with other drugs. As a result, many clinicians in Brazil have prescribed azithromycin in an attempt to combat or minimize the effects of COVID19. Aim: This study analyzed the sales data of the main antibiotics prescribed in Brazil to verify the change in consumption trends of these drugs during the COVID-19 pandemic. Methods: This is an interrupted time series that analyzed antimicrobial sales data between January 2014 and July 2021, publicly accessible information obtained from the Brazilian government’s website. Monthly means of “defined daily doses of DDDs” (DDDs per 1,000 inhabitants per day) of antibiotics were compared by analysis of variance, followed by the Dunnett Multiple Comparisons Test. Monthly trend changes in antibiotic use were verified using Joinpoint regression. Results: Amoxicillin (31.97%), azithromycin (18.33%), and cefalexin (16.61%) were the most sold antibiotics in Brazil during the evaluation period. Azithromycin consumption rose from 1.40 DDDs in February 2020 to 3.53 DDDs in July 2020. Azithromycin sales showed a significant increase in the pandemic period [Monthly Percent Change (MPC) 5.83%, 95% 1.80; 10.00], whereas there was a fall in amoxicillin sales (MPC −9.00%, 95% CI −14.70; −2.90) and cefalexin [MPC-2.70%, 95% (CI −6.30; −1.10)] in this same period. Conclusion: The COVID-19 pandemic changed the pattern of antibiotic consumption in Brazil, with a decrease in the use of amoxicillin and cefalexin and an increase in the consumption of azithromycin.

Determination of the standardized ileal digestible calcium requirement of male Arbor Acres Plus broilers from day 11 to 24 post-hatch

Male broilers (n = 576) were obtained and allocated to 96 cages with 6 birds per cage. From hatch to d 24, 16 randomly allocated cages were fed a nutrient adequate reference diet. The reference diet was formulated to contain 0.96 and 0.87% total Ca from hatch to d 10 and d 11 to 24, respectively. The remaining 80 cages were fed a nutrient adequate diet, formulated to contain 0.50% standardized ileal digestible (SID) Ca from hatch to d 10. On d 11, five diets containing 0.56, 0.46, 0.36, 0.26, or 0.16% SID Ca were randomly assigned to 16 cages per diet. Means were separated using polynomial contrasts and Dunnett's Multiple Comparison tests. From hatch to d 10, birds fed diets formulated to contain 0.50% SID Ca gained more (P < 0.05) compared with birds fed the reference diet. From d 11 to 24, there was no effect of diet on feed intake or BW gain. Birds fed 0.46 or 0.16% SID Ca were less (P < 0.05) efficient compared with birds fed the reference diet. Birds fed 0.26 or 0.36% SID Ca were more efficient compared with birds fed all other levels of SID Ca (quadratic, P < 0.05). Tibia ash percent was greatest in birds fed 0.56 or 0.46% SID Ca, decreased (quadratic, P < 0.05) as dietary SID Ca decreased and was lower (P < 0.001) in birds fed the 0.26 or 0.16% SID Ca diets compared with birds fed the reference diet. Apparent ileal digestibility (AID) of Ca or retention of P was greater (P < 0.05) in birds fed the diets formulated using SID Ca compared with birds fed the reference diet. Apparent P retention was greatest in birds fed 0.36% SID Ca (quadratic, P < 0.05). Regression equations developed using P retention, AID of Ca, and percent tibia ash estimated the SID Ca requirement of 11 to 24-d-old broilers was 0.380, 0.488, and 0.515%, respectively. This corresponds to a SID Ca to available P ratio of 0.86 to 1.17.

Effect of Mouthwash on Release of Elements from Dental Casting Alloys: An In-vitro Study

Introduction: Mouthwashes are regularly prescribed as a maintenance phase of treatment by the dentist. The pH of the mouthwash can enhance the elemental release from the cemented prosthesis. Aim: To evaluate the effect of two types (Oral-B and Listerine) of mouthwashes on the release of elements from Nickel-Chromium (Ni-Cr) dental alloys. Materials and Methods: An in-vitro comparative study was conducted in the Department of Prosthodontics, School of Dental Sciences, Karad, Maharashtra, India, from April 2018 to April 2022. A total of 90 crowns were fabricated using three different brands of dental casting alloys (DCA) using lost wax technique. The crowns were immersed into two different mouthwashes (Oral-B and Listerine) in polypropylene test tubes, and then incubated at 37°C to simulate the oral temperature for 1, 4, and 7 days. The leaching of the elements from the fabricated crowns into the mouthwashes was analysed using inductively coupled plasma mass spectrometry (ICP-MS). Data analysis was performed using the t test and repeated measure Analysis of Variance (ANOVA) followed by post hoc Dunnett Multiple Comparisons Test. Results: The Ni and Cr levels in the mouthwashes were significantly elevated in brand I (Ruby, Japan Dentistry Products Pvt. Ltd.), brand II (Dentmark, R&amp;D Impex International), and brand III (BEGO, GmbH &amp; Co KG). Presence of Ni released from all the three brands of DCA in mouthwash over a period of 1, 4, and 7 days was detected to be in the range of 16 to 39.9 µg/L. Cr released from all the three brands of DCA in mouthwash over a period of 1, 4, and days was detected to be in the range of 10.3 to 42.8 µg/L. Conclusion: The Ni and Cr were released in the mouthwash and the release increased with time. The acidic pH of Listerine adds to the increased release of Ni and Cr.

P-004: Pharmacological inhibition and depletion strategies for SLAMF7 CAR-T cells in multiple myeloma

<h3>Background</h3> Recent and soon-to-expect approvals have significantly increased the number of patients treated with CAR-T cells and clinical responses can be very impressive. However, side effects including cytokine storm, neurotoxicity or on-target/off-tumor toxicity can be challenging to manage. Further, malignant transformation of piggyBac modified CAR-T cells was recently reported in a phase I clinical trial as a potentially life threatening event. Reliable strategies to control or even deplete CAR-T cells are thus urgently needed. <h3>Methods</h3> We generated SLAMF7 CAR-T cells as previously described and exposed the cells to clinically relevant doses of the following drugs: dexamethasone, fludarabine, mafosfamide, dasatinib, cetuximab, and belantamab-mafodotin. After defined time periods, we evaluated absolute numbers of CAR-T cells and their effector functions against myeloma cell lines (cytotoxicity, cytokine secretion, proliferation). Statistical analysis was performed using one-way ANOVA, followed by Dunnett's multiple comparisons test if appropriate. The study was approved by the local ethics committee. <h3>Results</h3> Dexamethasone, used in the setting of severe CRS or ICANS, minimally decreased T cell numbers and cytokine release, but did not interfere with lytic capacity of the CAR-T cells. Fludarabine and the pre-activated cyclophosphamide analog mafosfamide, both drugs with potent lymphodepleting properties, significantly depleted numbers of CAR-T cells, reduced antigen-specific proliferation and led to an overall decrease in target cell elimination, even if specific lysis and cytokine release of the few surviving cells was preserved. Of note, fludarabine and mafosfamide similarly depleted unmanipulated T cells. Dasatinib, previously identified as a potent inhibitor of intracellular CAR signaling, had no impact on absolute T cell numbers, however lysis of myeloma cells, cytokine secretion and antigen-specific proliferation were entirely blocked. Evaluation of selective elimination of CAR-T cells with cetuximab, targeting the EGFR transduction marker of our CAR, showed modest CAR-T cell depletion in an ADCC assay. This was potentially related to competing natural killer cell elimination by the SLAMF7 CAR-T cells. To circumvent the problem of impaired ADCC, T cells were genetically engineered to express BCMA to enable simultaneous depletion of T cells and myeloma cells. Exposure to the BCMA-directed antibody-drug conjugate belantamab-mafodotin resulted in potent elimination of BCMA-positive T cells while unmanipulated cells were largely spared. <h3>Conclusions</h3> In aggregate, our data show that the different drugs can have justification in different clinical settings of CAR-T cell toxicity. While dasatinib and dexamethasone mainly act on T cell function, fludarabine and mafosfamide effectively deplete T cells in an unselective manner. Targeted elimination of CAR-T cells with antibodies can be augmented further by the use of antibody-drug-conjugates.

PD48-09 COMPARISON OF NON-INVASIVE MEASUREMENTS OF PENILE CURVATURE IN PEYRONIE'S DISEASE

You have accessJournal of UrologySexual Function/Dysfunction: Peyronie's Disease (PD48)1 Sep 2021PD48-09 COMPARISON OF NON-INVASIVE MEASUREMENTS OF PENILE CURVATURE IN PEYRONIE'S DISEASE Rubens Pedrenho Neto, Bruno C. G. Nascimento, Cristovão Machado Barbosa Neto, Jose de Bessa Junior, Miguel Srougi, and William Carlos Nahas Rubens Pedrenho NetoRubens Pedrenho Neto More articles by this author , Bruno C. G. NascimentoBruno C. G. Nascimento More articles by this author , Cristovão Machado Barbosa NetoCristovão Machado Barbosa Neto More articles by this author , Jose de Bessa JuniorJose de Bessa Junior More articles by this author , Miguel SrougiMiguel Srougi More articles by this author , and William Carlos NahasWilliam Carlos Nahas More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002070.09AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Peyronie’s disease (PD) is a fibrotic disorder of the tunica albuginea, leading to penile deformities. Non-invasive estimates of penile deformities are practical, but have low accuracy when compared to objective measurements with induced erection and a goniometer. Our aim was to evaluate the agreement between different non-invasive methods of measuring penile curvature and the objective measurement. METHODS: We retrospectively reviewed medical records of patients who underwent surgery in a sexual medicine tertiary center. During the clinical visit, pre-operatively, a set of different non-invasive estimates were performed including asking the patient to explain their primary curvature with a goniometer, a drawing of the penis and a malleable implant prosthesis to be molded. Also, at home photography were required with maximal erection. The agreement of those methods and intra-operative measurement using a goniometer after saline induced erection was analyzed. A tolerable error was considered if a difference smaller than 10o or 20% of the gold standard measurement was found. RESULTS: The sample consisted of 69 men with a mean age of 62 ± 6 years old. The direction of the curvature was dorsal in 59%, left in 35%, right in 5% and ventral in 1%. Mean curvature with of estimates with the goniometer was 57o (±31), drawing 60 o (±28), Prosthesis 60o (±25), Photography 57 o (±27), and mean intra-operative measurement was 51 o (± 24). Overall precision considering the above mentioned tolerable error definition was 45%, and the single non-invasive method with better precision was Prosthesis with 46,5% fo concordance. There was no statistical difference between them (p=0.99) for concordance and disagreement by the Dunnett's multiple comparisons test. CONCLUSIONS: In our comparison study of 4 non-invasive curvature assessment methods, we found a low precision (approximately 45%), regardless of the chosen method. Formal curvature assessment with induced erection and a goniometer appears to be imperative. Source of Funding: None © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e841-e841 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Rubens Pedrenho Neto More articles by this author Bruno C. G. Nascimento More articles by this author Cristovão Machado Barbosa Neto More articles by this author Jose de Bessa Junior More articles by this author Miguel Srougi More articles by this author William Carlos Nahas More articles by this author Expand All Advertisement Loading ...