Abstract Glioblastoma multiforme is a tumor of the central nervous system. Focal Adhesion Kinase (FAK) and αB-crystalline are two proteins involved in glioblastoma development. In this study, we investigated whether the FAK/αB-crystalline interaction is important for glioblastoma cells, we aimed to investigate the interaction of these two proteins in the glioblastoma multiforme cell line U87-MG. Two peptides named FP01 peptide (derived from αB-crystalline) and FP02 peptide (derived from FAK) were synthesized for this study. Treatment of U87-MG with the peptides FP01 and FP02 in the concentration at 50 µM reduced the viability cellular to around 41% and 51%, respectively. Morphological alterations in the cells treated with the peptides when compared to the control were observed. This study suggests that the interaction between FAK and αB-crystalline is important for the viability of glioblastoma cells. system (FAK αBcrystalline αB crystalline development FAK/αBcrystalline FAKαBcrystalline FAK/αB U87MG. U87MG UMG U87 MG. MG U FP FP0 derived 5 41 51 51% respectively observed FAKαB U8 4

Abstract A novel, simple and sensitive high-performance liquid chromatography with fluorescence detection method was developed and validated for the characterization of the preclinical pharmacokinetics of melatonin under pregnant conditions. Plasma samples (25 µL) were treated with 30 µL of ethanol absolute (containing the internal standard, IS). After a centrifugation process, aliquots of supernant (5 µL) were injected into the chromatographic system. Compounds were eluted on a Xbridge C18 (150 mm x 4.6 mm i.d., 5 µm particle size) maintained at 30°C. The mobile phase consisted in a mixture of aqueous solution of 0.4% phosphoric acid and acetonitrile (70:30 v/v). The wavelengths were set at 305 nm (excitation) and 408 nm (emission) and the total analysis time was 8 min/sample. All validation tests were obtained with accuracy and precision, according to FDA guidelines, over the concentration range of 0.005-20 µg/mL. Pharmacokinetic study showed that melatonin systemic exposure increased from day 14, with a significant difference at 19 days of gestation compared to the control group. Our findings suggest a decreased metabolism of melatonin as result of temporary physiological changes that occur throughout pregnancy. However, other maternal physiological changes cannot be ruled out. novel highperformance high performance conditions 25 (2 3 containing standard IS. IS . IS) process ( system C C1 150 (15 46 4 6 4. id i d i.d. size 30C 30°C 04 0 0.4 7030 70 (70:3 v/v. vv v/v v v/v) excitation (excitation 40 emission (emission minsample min sample min/sample precision guidelines 0.00520 000520 0.005 20 005 0.005-2 µgmL µg mL µg/mL 14 1 group pregnancy However out 2 15 (1 i.d 0. 703 7 (70: 00520 0.0052 00052 0005 0.00 00 0.005- (70 0052 000 0.0 (7

Abstract The goal of this work is to identify new fatty acid-mimetic 99mTc-complexes to be used as myocardial imaging agents that allow studying heart abnormalities in high-risk patients. In this sense, we designed a fatty acid-mimetic substructure including an amide moiety that, among other properties, could improve myocardial residence time. A diamide with a chain length of 15 atoms and porting a 6-hydrazinonicotinyl (HYNIC) chelator, and an analog with a short carbon-chain, were prepared with convergent organic synthetic procedures and radiolabeled with 99mTc using tricine as the sole coligand. The in vivo proofs of concept were performed using healthy mice. The new 99mTc-complexes were obtained with adequate radiochemical purity. The lipophilicities were in agreement with the length of the chains. While both 99mTc-complexes showed uptake in the myocardial muscle, the designed radiopharmaceutical with the longest chain length had preferential target-uptake and target-retention compared to other complexes described in the bibliography. Further studies, involving imaging assays, synthetic modifications, and assay of new coligands for 99mTc-HYNIC complexes, are currently ongoing. acidmimetic acid mimetic 99mTccomplexes mTccomplexes mTc highrisk high risk patients sense properties time 1 6hydrazinonicotinyl hydrazinonicotinyl 6 HYNIC (HYNIC chelator carbonchain, carbonchain carbon chain, carbon-chain coligand mice purity chains muscle targetuptake target targetretention retention bibliography studies assays modifications 99mTcHYNIC mTcHYNIC ongoing

Abstract Prostate cancer (PCa) is a highly prevalent condition among men worldwide, resulting in reduced quality of life and increased costs to health systems due to hospitalization and death. This study aimed to explore and understand the evolution of PCa in Brazil from 2008 to 2018. Data were obtained from the National Health System Department of Informatics (DATASUS) using code C61 for malignant prostatic neoplasms. We presented the hospitalization and mortality rates in a temporal-, regional- and age-dependent manner. From 2008 to 2018, a year-dependent increase in hospital admissions due to PCa was reported in Brazil, in which the Southeast region showed the highest prevalence. Men aged ≥80 and those 70-79 years old had similar hospitalization rates, followed by men aged 60-69, 50-59, 40-49 and 30-39 years old. Similarly, an increase in deaths due to PCa was reported during this period, with the highest rates seen in the Southeast. Men aged ≥80 years had higher mortality rates, followed by those aged 70-79, 60-69, 50-59, 40-49 and 30-39 years old. The results obtained indicate an age- and region-dependent increase in PCa morbidity and mortality in Brazil overtime and may contribute to the ongoing discussion on the role and future perspective of the health care system in Brazil. (PCa worldwide death 200 2018 DATASUS (DATASUS C C6 neoplasms temporal, temporal , temporal- regional agedependent age dependent manner yeardependent year prevalence 80 ≥8 7079 70 79 70-7 6069, 6069 60 69, 69 60-69 5059, 5059 50 59, 59 50-59 4049 40 49 40-4 3039 30 39 30-3 Similarly period 7079, 79, regiondependent 20 201 8 ≥ 707 7 70- 606 6 60-6 505 5 50-5 404 4 40- 303 3 30- 2 60- 50-

Abstract Humans are exposed to natural compounds such as phytoestrogens primarily through diet and supplements. These compounds promote health by alleviating the symptoms and illnesses associated with menopause and arthritis. Diosgenin (DSG) occurs naturally in plants such as Dioscorea villosa (DV) and binds to estrogen receptors, so it may have similar effects to this hormone, including against arthritis. Thus, we investigated the effect of chronic treatment with dry extract of DV and its phytoestrogen DSG on ovariectomized mice with arthritis. We found that dry extract of Dioscorea villosa (DV) contains the phytoestrogen diosgenin (DSG) in its composition. Furthermore, arthritic mice treated with DV and DSG showed reduced neutrophil accumulation in the articular cartilage. Also, the dry extract of DV administered orally (v.o) did not alter the leukocyte count in the joints or promote changes in the reproductive tract. However, DSG altered these parameters, with possible beneficial effects by reducing symptoms related to reproductive aging. Thus, oral treatment with dry extract of DV and subcutaneous (s.c) treatment with DSG showed promise by acting against inflammation caused by arthritis and reducing symptoms in the reproductive tract due to menopause. supplements (DSG (DV receptors hormone Thus composition Furthermore cartilage Also v.o vo v o (v.o However parameters aging s.c sc s c (s.c

Abstract Medicines must be subject to physical, chemical, and biological analysis to guarantee their quality, safety, and effectiveness. Despite the efforts to ensure the reliability of analytical results, some uncertainty will always be associated with the measured value, which can lead to false decisions regarding conformity/non-conformity assessment. This work aims to calculate the specific risk of false decisions regarding conformity/non-conformity of acetaminophen oral solution dosage form. The acetaminophen samples from five different manufacturers (A, B, C, D, and E) were subject to an active pharmaceutical ingredient assay, density test, and dose per drop test according to the official compendia. Based on measured values and their respective uncertainties, the risk values were calculated using the Monte Carlo method implemented in an MS Excel spreadsheet. The results for two acetaminophen oral solution samples (C and D) provided an increased total risk value of false acceptance (33.1% and 9.6% for C and D, respectively). On the other hand, the results for the other three acetaminophen samples (A, B, and E) provided a negligible risk of false acceptance (0.004%, 0.025%, and 0.045% for A, B, and E, respectively). This indicates that measurement uncertainty is very relevant when a conformity assessment is carried out, and information on the risks of false decisions is essential to ensure proper decisions. physical chemical quality safety effectiveness conformity/nonconformity conformitynonconformity conformity/non non form A (A B D E assay compendia uncertainties spreadsheet 33.1% 331 33 1 (33.1 96 9 6 9.6 respectively. respectively . respectively) hand 0.004%, 0004 0 004 (0.004% 0025 025 0.025% 0045 045 0.045 out nonconformity conformitynon 33.1 3 (33. 9. 0.004% 000 00 (0.004 002 02 0.025 04 0.04 33. (33 0.004 (0.00 0.02 0.0 (3 0.00 (0.0 0. ( (0. (0

Abstract We developed poly-ε-caprolactone (PCL)-based nanoparticles containing D-α-tocopherol polyethylene glycol-1000 succinate (TPGS) or Poloxamer 407 as stabilizers to efficiently encapsulate genistein (GN). Two formulations, referred to as PNTPGS and PNPol, were prepared using nanoprecipitation. They were characterized by size and PDI distribution, zeta potential, nanoparticle tracking analysis (NTA), GN association (AE%), infrared spectroscopy (FT-IR), and differential scanning calorimetry (DSC). PNTPGS-GN exhibited a particle size of 141.2 nm, a PDI of 0.189, a zeta potential of -32.9 mV, and an AE% of 77.95%. PNPol-GN had a size of 146.3 nm, a better PDI than PNTPGS-GN (0.150), a less negative zeta potential (-21.0 mV), and an AE% of 68.73%. Thermal and spectrometric analyses indicated that no new compounds were formed, and there was no incompatibility detected in the formulations. Cellular studies revealed that Poloxamer 407 conferred less toxicity to PCL nanoparticles. However, the percentage of uptake decreased compared to the use of TPGS, which exhibited almost 80% cellular uptake. This study contributes to the investigation of stabilizers capable of conferring stability to PCL nanoparticles efficiently encapsulating GN. Thus, the PCL nanoparticle proposed here is an innovative nanomedicine for melanoma therapy and represents a strong candidate for specific pre-clinical and in vivo studies. polyεcaprolactone poly ε caprolactone PCLbased based Dαtocopherol D α tocopherol glycol1000 glycol 1000 glycol-100 TPGS (TPGS 40 . (GN) formulations PNPol nanoprecipitation distribution NTA, NTA , (NTA) AE%, AE (AE%) FTIR, FTIR FT IR (FT-IR) DSC. DSC (DSC) PNTPGSGN 1412 141 2 141. nm 0189 0 189 0.189 32.9 329 32 9 -32. mV 7795 77 95 77.95% PNPolGN 1463 146 3 146. 0.150, 0150 0.150 150 (0.150) 21.0 210 21 (-21. mV) 6873 68 73 68.73% formed However 80 Thus preclinical pre clinical glycol100 100 glycol-10 4 (GN (NTA (AE% (FT-IR (DSC 14 018 18 0.18 32. -32 779 7 77.95 015 0.15 15 (0.150 21. (-21 687 6 68.73 8 glycol10 10 glycol-1 (AE 1 01 0.1 -3 77.9 (0.15 (-2 68.7 glycol1 glycol- 0. - 77. (0.1 (- 68. (0. ( (0

Abstract There are a limited number of studies examining the effects of the pandemic on the daily lives of Turkish community pharmacists, and no research investigating the impact on the lives of Turkish hospital pharmacists has been found. This study aimed to examine the effects of the pandemic on the personal and professional lives of Turkish community pharmacists and hospital pharmacists. In this qualitative study design, a comprehensive set of interviews was conducted with a total of 13 community pharmacists and 7 hospital pharmacists, employing a semi-structured interview guide. Through thematic content analysis of the interviews, four main themes, 1) long-term impacts, 2) dealing strategies, 3) professional life impacts, 4) personal life impacts, have emerged for both community pharmacists and hospital pharmacists. In addition to the psychological impacts and supply chain issues commonly mentioned in the literature, the study revealed ongoing effects such as the inability to sell available products and economic difficulties. Also, the increased demand for over-the-counter products during the pandemic highlights the need for the government to develop policies to address this issue. found design 1 semistructured semi structured guide themes longterm long term 2 strategies 3 4 literature difficulties Also overthecounter over counter issue

Abstract This study attempted to describe the disease modifying therapies (DMTs) use patterns and related adverse events in multiple sclerosis (MS) patients assisted by the Brazilian Unified Health System (SUS). This is a cross-sectional study conducted at a reference center in the Midwestern Brazil. Demographic, socioeconomic, and clinical data, and adverse events associated with DMTs were collected. We observed 291 patients with a mean age of 41.4 years, and mostly (68.6%) women. Most patients (58.8%) were using first-line treatment. Fingolimod (29.9%) and beta interferons (23.7%) were the most used drugs. About 74.2% of patients used DMTs for more than six months. In 26.5% of patients, 238 adverse events were reported, 67.2% of which were mild and 32.8% of which were moderate. The most frequent adverse events were headaches (6.9%), myalgia (3.8%), and flu-like symptoms (3.1%). The proportion of adverse events proportion ranged from 17.3% (natalizumab) to 41.2% (dimethyl fumarate). Most MS patients treated by SUS used first-line DMTs. After adjustment, there was noticed that adverse events associated with DTMs are twice as likely to occur in users of the first-line treatment than other lines (OR 1.99, p=0.01). It is essential to develop DMTs safety monitoring strategies to promote their rational use. (DMTs (MS SUS. . (SUS) crosssectional cross sectional Brazil Demographic socioeconomic data collected 29 414 41 4 41. years 68.6% 686 68 6 (68.6% women 58.8% 588 58 8 (58.8% firstline first line 29.9% 299 9 (29.9% 23.7% 237 23 7 (23.7% drugs 742 74 2 74.2 months 265 26 5 26.5 reported 672 67 67.2 328 32 32.8 moderate 6.9%, 69 6.9% , (6.9%) 3.8%, 38 3.8% 3 (3.8%) flulike flu like 3.1%. 31 3.1% 1 (3.1%) 173 17 17.3 natalizumab (natalizumab 412 41.2 dimethyl fumarate. fumarate fumarate) adjustment OR 199 99 1.99 p=0.01. p001 p p=0.01 0 01 p=0.01) (SUS 68.6 (68.6 58.8 (58.8 29.9 (29.9 23.7 (23.7 74. 26. 67. 32. 6.9 (6.9% 3.8 (3.8% 3.1 (3.1% 17. 19 1.9 p00 p=0.0 68. (68. 58. (58. 29. (29. 23. (23. 6. (6.9 3. (3.8 (3.1 1. p0 p=0. (68 (58 (29 (23 (6. (3. p=0 (6 (5 (2 (3 p= (

Abstract In Brazil, hepatitis C virus (HCV) treatment has gone through several stages until it reached the current schemes. This study aimed to evaluate the clinical, epidemiologic, and effectiveness of treatment with second-generation direct-acting antivirals (DAAs) for HCV-infected patients in southern Brazil. Medical records of a reference center of Uruguaiana-RS/Brazil were evaluated from December 2017 to August 2020. Data collected included demographics, disease severity and comorbidities, genotype, viral load, medications, treatment side effects, and sustained virological response (SVR). A multivariate linear regression model was developed to identify the factors associated with non-sustained virological response. The HCV-infected patients were predominantly male, white, Genotype 1, with initial liver fibrosis (F0/F1), and untreated patients. Laboratory parameters of the liver profile showed significant improvement after completion of treatment with second-generation DAA for 12 weeks. Significant results were found in the Genotype where 7.3 and 6.0 times more chances of Genotype 1a and Genotype 3, respectively, to present detectable SVR compared to Genotype 1b. Variable cardiovascular comorbidity also indicated a higher risk of absence of SVR compared to individuals without this comorbidity. Treatment with second-generation DAAs can contribute to the Brazilian hepatitis C elimination plan proposed by the World Health Organization. Brazil HCV (HCV schemes clinical epidemiologic secondgeneration second generation directacting direct acting (DAAs HCVinfected infected UruguaianaRS/Brazil UruguaianaRSBrazil Uruguaiana RS/Brazil RS 201 2020 demographics comorbidities genotype load medications effects SVR. . (SVR) nonsustained non male white 1 F0/F1, F0F1 FF F0/F1 , F0 F1 F (F0/F1) weeks 73 7 3 7. 60 6 0 6. respectively 1b b Organization UruguaianaRS RSBrazil 20 202 (SVR F0F F0/F (F0/F1 2 (F0/F

Abstract The pharmaceutical industry is mostly dedicated to the production of drugs in solid forms such as tablets and capsules with active ingredients and excipients in the same physical state. Regarding the regulatory requirements for the analytical development and implementation of solid-state analyses, internationally recognized pharmaceutical compendia play an important role. However, the information contained in the various general chapters and monographs differs from each other, especially in countries that have national pharmacopeias, such as Brazil. Thus, the main objective of this work is to critically evaluate, based on the technical-scientific literature, the harmonization of the United States, European, British, Japanese and Brazilian pharmacopeias with respect to the following analytical techniques used for solid-state characterization: X-ray powder diffraction, differential scanning calorimetry, thermogravimetric analysis, and infrared and Raman spectroscopies. The working principle of each analytical technique, as well as the methodological parameters that impact the implementation of the analyses, are detailed. The results indicate that, in terms of solid-state characterization analysis, the adoption of unified scientific standards and principles is not yet a reality for all compendia. Additionally, the lack of harmonization between BrazP and the other compendia is especially significant, considering that ANVISA is an ICH member, the main entity responsible for promoting harmonization. state solidstate analyses role However Brazil Thus evaluate technicalscientific technical literature States European British Xray X ray diffraction calorimetry analysis spectroscopies technique detailed Additionally significant member

Abstract Valproic acid (VPA) is an effective and inexpensive anticonvulsant commonly prescribed as an antiepileptic. However, there is a percentage of susceptible patients who experience unwanted effects, such as weight gain and fatty liver. Ivermectin (IVM), a versatile antiparasitic, has shown potential in the treatment of non-alcoholic fatty liver disease. In this study, the potential hepatoprotective effect of ivermectin on valproic acid-induced hepatotoxicity was investigated in rats. VPA (250 mg/kg/day) was orally administered for 14 consecutive days to induce hepatocellular damage in adult Wistar rats, after which the animals were treated with a subcutaneous injection of IVM (0.4 and 1.3 mg/kg/day) for another five days. Animals receiving VPA developed obesity and microvesicular and macrovesicular steatosis. They also had increased levels of plasma triglycerides and aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase enzymes. IVM treatment significantly decreased serum triglyceride and cholesterol levels and attenuated valproic acid-induced hepatic lipid accumulation in rats. These results were confirmed by histopathological examination. (VPA antiepileptic However effects IVM, , (IVM) antiparasitic nonalcoholic non alcoholic disease study acidinduced induced rats 250 (25 mg/kg/day mgkgday mg kg day 1 0.4 04 0 4 (0. 13 3 1. steatosis enzymes examination (IVM 25 (2 0. (0 2 (

Abstract Candida albicans is the main fungus responsible for acute and recurrent vaginal infections. Low adherence to treatment is due to dosing frequency and available topical medications. Thus, we developed mucoadhesive tablets based on Schinopsis brasiliensis Engler extract composed of Carboxymethylcellulose (F1), Hydroxypropylmethylcellulose (F2), or Pluronic (F3), against C. albicans . The extract metabolites determination showed presence of polyphenols (15 μg.mg-1), flavonoids (5.51 μg.mg-1), and tannins (4.80 μg.mg-1). The in vitro antifungal activity was performed by broth microdilution method. Quality control tests were performed according international pharmacopeias. Regarding the mucoadhesion test, in terms of the force expressed between the tablet:mucin disc, the average values were approximately 0.1267 N (F1), 0.0411 N (F2) and 0.814 N (F3), while in the tablet:vaginal mucosa, the average values were approximately 0.0294 N (F1), 0.0166 N (F2) and 0.0365 N (F3). The dry extract showed a minimum inhibitory concentration of 62.5 μg.mL-1. In evaluating the in vitro release of tablets, polymers derived from cellulose released 100% in less than 4 hours, while Pluronic released around 56.3% in 72 hours. It is concluded that F1 has greater mucoadhesion while F3 has a modified release rate, providing promising results for future application of the formulation in the clinical assay. infections medications Thus F1, F , (F1) F2, F2 F3, (F3) C 15 (1 μg.mg1, μgmg1 μgmg μg.mg 1 μg mg μg.mg-1) 5.51 551 5 51 (5.5 4.80 480 80 (4.8 μg.mg1. method pharmacopeias test tabletmucin tablet mucin disc 01267 0 1267 0.126 00411 0411 0.041 (F2 0814 814 0.81 tabletvaginal mucosa 00294 0294 0.029 00166 0166 0.016 00365 0365 0.036 F3. 625 62 62. μg.mL1. μgmL1 μgmL μg.mL 1. mL μg.mL-1 100 hours 563 56 3 56.3 7 rate assay (F1 (F3 ( mg1 μg.mg1 μg.mg-1 5.5 55 (5. 4.8 48 8 (4. 0126 126 0.12 0041 041 0.04 (F 081 81 0.8 0029 029 0.02 0016 016 0.01 0036 036 0.03 6 mL1 μg.mL1 μg.mL- 10 56. μg.mg- 5. (5 4. (4 012 12 0.1 004 04 0.0 08 0. 002 02 001 01 003 03 00

Abstract Cutaneous melanoma, a highly aggressive skin cancer, is the leading cause of death among skin neoplasms. Hence, this study focused on developing a 3,3'-diindolylmethane (DIM)-loaded pomegranate oil (PO) nanocapsule suspensions and their potential effects on melanoma cells. Nanocapsule suspensions were prepared by the interfacial deposition method (Eudragit ® RS100). Medium-chain triglyceride (MCT) nanocapsules were produced for comparison. The of nanocapsule suspensions showed nanometric size (<180 nm), low polydispersity index (<0.12) (as measured by dynamic light scattering), positive zeta potential (tested by capillary microelectrophoresis), pH below 5.0 (as measured by potentiometry), DIM content, and nearly 100% encapsulation efficiency (evaluated by HPLC). The formulations were considered non-irritating (according to HET-CAM) regardless of the core constitution. Notably, PO-based nanocapsules showed a superior performance compared to the MCT-containing formulation in delaying DIM light degradation (tested by UVC radiation), lowering genotoxicity (determined by Allium cepa ), and increasing in vitro antioxidant action (measured by ABTS and FRAP). Additionally, the PO-based formulation demonstrated higher in vitro antitumoral activity in B16F10 than MCT nanocarriers (IC 50 of 6.49 and >24 µg/mL, respectively), and antimetastatic capacity by reducing B16F10 cell adhesion, colonization, and migration. Based on these results, this study suggests that the nanoencapsulation of DIM into PO nanocapsules may be a promising approach in melanoma treatment. cancer neoplasms Hence 3,3diindolylmethane 33diindolylmethane diindolylmethane 3,3 3 DIMloaded loaded (PO cells Eudragit RS100. RS100 RS . RS100) Mediumchain Medium chain (MCT comparison <180 180 (<18 nm, nm , nm) <0.12 012 0 12 (<0.12 as scattering, scattering scattering) tested microelectrophoresis, microelectrophoresis microelectrophoresis) 5 5. potentiometry, potentiometry potentiometry) content 100 evaluated HPLC. HPLC HPLC) nonirritating non irritating according HETCAM HET CAM HET-CAM constitution Notably PObased based MCTcontaining containing radiation, radiation radiation) determined ) FRAP. FRAP FRAP) Additionally BF B F B16F1 IC 649 6 49 6.4 24 >2 µgmL µg mL µg/mL respectively, respectively respectively) adhesion colonization migration results treatment 3diindolylmethane 33 3, RS10 <18 18 (<1 <0.1 01 1 (<0.1 10 B16F 64 4 6. 2 > RS1 <1 (< <0. (<0. < ( <0 (<0

Abstract Caffeine consumption by academics, especially those in medical school, is done indiscriminately due to the need to reduce fatigue during their activities and, consequently, increasing their performance. Although it seems to be a harmless practice, it can be related to some health problems. Among these other damages are: arrhythmias, urinary incontinence, loss of appetite, nausea, increased blood pressure, insomnia and sudden death. In this context, this work evaluated cardiovascular damage associated to long-term use of caffeine based on clinical evaluation of students aged 18 years or older in a private medical college in Espírito Santo. Analysis was performed through a brief questionnaire followed by assessment of heart rate and blood pressure. The work was performed with 26 men and 69 women and an average of 70% of the population claimed to consume caffeine. It was also seen that among the students who had the greatest changes in relation to blood pressure were those who were in the 1st and 3rd years of the course. In short, no significant changes were observed in relation to caffeine consumption throughout graduation or cardiovascular impacts associated with its use. However, the evaluated individuals are mostly young and the study follow-up time was short. academics school consequently performance practice problems arrhythmias incontinence appetite nausea death context longterm long term 1 Santo 2 6 70 st rd course short However followup follow up 7