Diagnostic accuracy and the first genotype-phenotype correlation in glycogen storage disease type V.

BACKGROUND: Glycogen storage disease type V (GSDV) is an autosomal recessive metabolic condition caused by pathogenic PYGM variants. This is an underdiagnosed condition as it presents with exercise intolerance in children. We reviewed the GSDV cases of a tertiary hospital center to assess diagnostic timing/accuracy, as well as potential clinical/analytical predictors of such factors. METHODS: We retrospectively reviewed all GSDV cases with follow-up in both Pediatric and Adult Metabolic Diseases consultations. We included 28 cases and assessed their hospital record for clinical information. RESULTS: Over 90% of our cases had late diagnoses, with more than 50% being diagnosed in adulthood despite symptom onset in preschool (very late diagnosis). Diagnostic age was lower in patients exhibiting myoglobinuria. Interestingly, patients with a positive family history of GSDV had similar rates of very late diagnoses, likely since the index case was already detected very late in life. Finally, we observe that the R50* variant is associated with increased myoglobinuria and CK elevation, in a dosage-dependent manner. CONCLUSION: We concluded that GSDV is severely underdiagnosed, and that some clinical and analytical aspects of the condition can be more indicative of this diagnosis. Furthermore, we propose for the first time a genotype-phenotype correlation in GSDV. IMPACT: GSDV is a pediatric-onset metabolic disorder that is mostly diagnosed late in the adult age and commonly misdiagnosed. We observed the first genotype-phenotype correlation in GSDV, regarding the common R50* variant. Awareness of GSDV for pediatricians and the overall medical community is vital.

Professional karate-do and mixed martial arts fighters present with a high prevalence of temporomandibular disorders.

BACKGROUND/AIM: Facial trauma in sports has been associated with temporomandibular disorders. Because of the intensity and duration of training needed for elite-level competitions, high-performance athletes can have two to five times more traumatic injuries than recreational athletes. This study aimed to investigate the prevalence of temporomandibular disorders in high-performance martial arts fighters and compare it with the prevalence in recreational athletes and non-athletes. MATERIAL AND METHODS: The Research Diagnostic Criteria for Temporomandibular Disorders was used to diagnose and classify professional karate-do practitioners (group I; n = 24), amateur karate-do practitioners (group II; n = 17), high-performance mixed martial arts fighters (group III; n = 13), and non-athletes (n = 28). The groups were compared with the chi-square test and tested for the difference between two proportions using a significance level of 5% (P < 0.05). RESULTS: The prevalence of temporomandibular disorders in groups I (54.2%; P = 0.003) and III (61.5%; P = 0.002) was significantly higher than in group IV (14.3%). The prevalence in group II was similar to that in group IV (P > 0.05). A diagnosis of arthralgia from disk displacement was made more frequently in groups I (45.8%; P = 0.013) and III (38.5%; P = 0.012) than in group IV (7.1%). The chronic pain associated with TMD was low intensity and low disability. CONCLUSIONS: While there was a high prevalence of temporomandibular disorders in the professional athletes in our study, the prevalence of the condition in recreational athletes was similar to that in individuals who did not practice martial arts.

Cleaning and disinfection of the root canal system provided by four active supplementary irrigation methods.

This in vitro study evaluated the bacterial reduction provided by the EndoActivator (EA), Easy Clean (EC), passive ultrasonic irrigation (PUI), and XP-Endo Finisher. Eight-four mesial roots of mandibular first molars were instrumented, inoculated with Enterococcus faecalis, and divided into four groups (n. 20). Bacterial reduction in the main canals and dentinal tubules were respectively determined by MTT assays and Live/Dead BackLight technique through confocal laser scanning microscopy (CLSM) at 50, 100, and 150 µm in-depth (n. 10 per group). Statistical analyses were conducted following a significance level of 95% (P < 0.05). A significant statistical difference was just identified between XPF and EC in the main canals. In the dentinal tubules from the main root canals, at 100 and 150 µm in-depths, significant statistical differences were only observed between XPF and EC (P = 0.027) for the former and between XPF and EC (P = 0.011) and XPF and PUI (P = 0.021) for the latter. In the dentinal tubules from the isthmus, at 100 µm in-depth, statistically relevant differences did occur between XPF and EC (P = 0.038) and EC and EA (P = 0.029). At 150 µm in-depth, these differences were only significant by comparing XPF and PUI (P = 0.025) and XPF and EC (P = 0.036). Although no irrigation method could thoroughly disinfect the RCS, bacterial reduction indexes were generally better after using XPF.

Clinical and Laboratory Findings on Glycogen Storage Disease Type V: Results from a Retrospective Observational Study in a Tertiary Hospital.

Introduction - Glycogen storage disease type V (GSDV, MIM #232600) is an autosomal recessive metabolic myopathy caused by pathogenic variants in the PYGM gene. The characteristic symptoms of exercise intolerance, myalgia, and cramps, which improve after a few minutes of rest, are frequently unrecognized in affected children. When there is clinical suspicion, the initial approach with a forearm exercise test has diagnostic value by detecting low post-exercise plasma lactate-to-ammonia ratio values. The diagnostic algorithm is followed by genetic testing if the results suggest myophosphorylase deficiency. Methods - This was a retrospective observational study conducted based on reviewing medical records of patients with GSDV in a tertiary hospital. We assessed demographic variables, including the timing of onset and diagnosis, relevant clinical characteristics, and whether genetic testing was performed, including its results. Results/Case Report - Our goal was to review the GSDV cases in our center to assess our cohort's diagnostic timing and clinical and genetic characteristics. We identified 28 patients from 24 families, three with consanguinity. The mean age at the time of the study was 43 years. While most (26/28; 93%) recalled their first symptoms in childhood/adolescence, only 25% (7/28) were diagnosed then. All patients had exercise intolerance and CK elevation, while about half reported the second wind phenomenon. Genetic testing was performed in 22 patients, revealing biallelic PYGM variants (9 homozygous, 13 compound heterozygous) as the most common (p.R50*). Conclusion - GSDV is rare and presents in the pediatric age, with subtle manifestations often underestimated for decades. A late diagnosis may negatively impact the psychosocial development of affected children. It is essential to recognize some unique features that facilitate diagnosis: history of exercise intolerance, the second wind sign, and high resting serum CK levels. Identifying the disease-causing variants in PYGM is currently the gold standard for diagnosis as it is less invasive than performing a muscle biopsy, and may promptly diagnose the condition and avoid wrongful labelling of patients.

Leukocyte Imbalances in Mucopolysaccharidoses Patients.

Mucopolysaccharidoses (MPSs) are rare inherited lysosomal storage diseases (LSDs) caused by deficient activity in one of the enzymes responsible for glycosaminoglycans lysosomal degradation. MPS II is caused by pathogenic mutations in the IDS gene, leading to deficient activity of the enzyme iduronate-2-sulfatase, which causes dermatan and heparan sulfate storage in the lysosomes. In MPS VI, there is dermatan sulfate lysosomal accumulation due to pathogenic mutations in the ARSB gene, leading to arylsulfatase B deficiency. Alterations in the immune system of MPS mouse models have already been described, but data concerning MPSs patients is still scarce. Herein, we study different leukocyte populations in MPS II and VI disease patients. MPS VI, but not MPS II patients, have a decrease percentage of natural killer (NK) cells and monocytes when compared with controls. No alterations were identified in the percentage of T, invariant NKT, and B cells in both groups of MPS disease patients. However, we discovered alterations in the naïve versus memory status of both helper and cytotoxic T cells in MPS VI disease patients compared to control group. Indeed, MPS VI disease patients have a higher frequency of naïve T cells and, consequently, lower memory T cell frequency than control subjects. Altogether, these results reveal MPS VI disease-specific alterations in some leukocyte populations, suggesting that the type of substrate accumulated and/or enzyme deficiency in the lysosome may have a particular effect on the normal cellular composition of the immune system.

Niemann-Pick Type B: A Rare Cause of Interstitial Lung Disease.

Niemann-Pick disease (NPD) is a rare lysosomal storage disease (LSD) with multisystemic involvement. The disease is heterogeneous and classified into three subtypes: type A and B result from deficient acid sphingomyelinase activity and leads to the accumulation of sphingomyelin and type C is a genetically different disease resulting from defective intracellular trafficking of cholesterol with accumulation of glycosphingolipids. Type A is generally a neurodegenerative disease and is fatal in infancy. Type B is a less severe form characterized by pulmonary involvement, hepatosplenomegaly, hyperlipidemia and most patients live into adulthood. In type C, clinical presentation is dominated with neurological involvement. Pulmonary involvement occurs in all three types of Niemann-Pick but most frequently in type B. Clinical manifestations range from a lack of symptoms to respiratory failure, and respiratory symptoms are usually mild with recurrent cough, dyspnoea on exertion and recurrent respiratory infections. Interstitial lung disease (ILD) is the most prominent feature with slow progression, characterized by worsening pulmonary function tests. In recent years, enzyme replacement therapy has shown promising results in clinical trials, such as improvement in organomegaly and pulmonary involvement with the potential to improve patients' lives. We present three cases of NPD with pulmonary involvement, each exhibiting a different pattern of ILD and evaluate therapeutic options.

Hunter Syndrome: The Phenotype of a Rare Storage Disease.

Hunter syndrome is a rare lysosomal storage disorder with systemic involvement that occurs over time. Affected patients have coarse facial features, growth retardation with short stature, and skeletal deformities called dysostosis multiplex; joint stiffness, progressive mental retardation, and organomegaly are some of the clinical signs. It ranges from mild to severe manifestations and the distinction between them is related to neurological involvement. Cardiac and respiratory failure is commonly the cause of early death (before adulthood) for severe forms, but those with attenuated forms who have normal cognitive development can survive until late adulthood. Treatment with enzyme replacement therapy is available and can improve the prognosis of this disease. The authors present a case of a 36-year-old male with Hunter syndrome to show not only the clinical features typical of this multisystemic disease that should alert to a prompt investigation but also to remind that treatment must start as early as possible to reach the best outcome. Management of this disease is typically challenging and requires a multidisciplinary approach.

Glutaric Aciduria Type 2 Presenting in Adult Life With Hypoglycemia and Encephalopathic Hyperammonemia.

Glutaric aciduria type 2 is a rare inborn disease of fatty acid metabolism. The clinical manifestation of this disease is heterogeneous and ranges from severe neonatal forms to mild late-onset forms. We present a case of a previously healthy 22-year-old woman with unexplainable hypoglycemia and encephalopathic hyperammonemia. Acylcarnitine profile and organic acids analysis were compatible with glutaric aciduria type 2. On suspicion of this disease, the patient started supplements with carnitine and riboflavin, along with hemodialysis with a complete recovery. The genetic test confirmed the diagnosis. Glutaric aciduria type 2 has no cure and the metabolic decompensation can be a severe event, but treatable and preventable, if this pathology gets recognized.

Is (R)-ketamine a potential therapeutic agent for treatment-resistant depression with less detrimental side effects? A review of molecular mechanisms underlying ketamine and its enantiomers.

Approximately one-third of individuals with major depressive disorder are resistant to conventional antidepressants (i.e., monoamine-based therapies), and, even among respondents, a proper therapeutic effect may require weeks of treatment. Ketamine, a racemic mixture of the two enantiomers, (R)-ketamine and (S)-ketamine, is an N-methyl-d-aspartate receptor (NMDAR) antagonist and has been shown to have rapid-acting antidepressant properties in patients with treatment-resistant depression (TRD). Although (R)-ketamine has a lower affinity for NMDAR, it presents greater potency and longer-lasting antidepressant properties, with no major side effects, than racemic ketamine or (S)-ketamine in preclinical findings. Thereby, ketamine and its enantiomers have not only an antagonistic effect on NMDAR but also a strong synaptogenic-modulatory effect, which is impaired in TRD pathophysiology. In this review, we summarize the current evidence regarding the modulation of neurotransmission, neuroplasticity, and neural network activity as putative mechanisms of these rapid-acting antidepressants, highlighting differences on intracellular signaling pathways of synaptic proteins such as mammalian target of rapamycin (mTOR), extracellular signal-regulated kinase (ERK) and brain-derived neurotrophic factor (BDNF). In addition, we discuss probable mechanisms involved in the side effects of ketamine and its enantiomers.

Role of central obesity on pain onset and its association with cardiovascular disease: a retrospective study of a hospital cohort of patients with osteoarthritis.

OBJECTIVES: To determine the role of central obesity (CO) in the onset and severity of joint pain and in predicting cardiovascular disease (CVD) in subjects affected with osteoarthritis (OA). DESIGN: Retrospective analysis on the onset of OA joint pain and CO. Waist circumference (WC), Waist-to-height ratio andwaist-to-hip ratio (WHR) were measured at the interview and defined according to the WHO criteria. Cross-sectional analyses on the association of comorbidities, including CVD, pain severity (number of joints and pain score) and CO. SETTINGS AND PARTICIPANTS: Medical records and interviews of a hospital cohort study of 609 patients with OA. Analyses included analysis of variance, mean differences (MDs), SE and logistic regression. Areas under the receiver operating characteristic curve (AUROC) compared the predictive value of the sex-specific CVD models. OUTCOME MEASURES: Onset of OA joint pain (years) and severity according to body mass index (BMI) and WC categories. Predictive value of WC for CVD by sex. Education level, disability, smoking and alcohol use were used to adjust the analysis. RESULTS: Subjects with OA and CO by WHR started 2 years earlier with pain symptoms and had more joints affected than those without CO (MD=1.96 years, SE=0.95, p=0.04 and MD=0.32, SE=0.15 and p=0.04, respectively). Age and hypertension were associated with CVD in both genders, and NSAIDs use only in males. In addition, respiratory disease, hypercholesterolaemia, stairs difficulty, a wider WC and obesity were significant risk factors in females, improving 12.7% in the prediction of CVD cases, compared with only age and BMI (AUROCC=0.793 and 0.666, respectively, p=0.03 for the difference between AUROCs). CONCLUSION: CO is associated with the onset of joint pain, and all pain analysed variables. CO has a role in CVD in women affected with OA and might help predict CVD cases.

Demographic and epidemiological characteristics of pregnant and postpartum women who died from severe acute respiratory syndrome in Brazil: A retrospective cohort study comparing COVID-19 and nonspecific etiologic causes.

The objective of this study is to compare the demographic characteristics and symptoms in pregnant and postpartum women who died from Severe Acute Respiratory Syndrome (SARS) caused by COVID-19 or by nonspecific cause in different states of Brazil. This is a retrospective cohort study and the analysis was conducted on SARS death records between 02/16/2020 and 04/17/2021, obtained from the Information System for the Epidemiological Surveillance of Influenza (Sistema de Informação da Vigilância Epidemiológica da Gripe, SIVEP-Gripe). Pregnant and postpartum women, aged between 10 and 55 years, who died from SARS, were included and classified into two groups: SARS due to confirmed COVID-19 or SARS due to nonspecific cause. The cases were analyzed according to the women's demographic and epidemiological characteristics, clinical symptoms, risk factors and disease evolution. As results, 19,333 pregnant and postpartum women were identified. From these, 1,279 died (1,026 deaths from COVID-19 and 253 deaths from SARS with nonspecific cause). The groups showed significant differences in age, education, race, and occurrence of obesity and chronic lung disease. The group of women who died from confirmed COVID-19 presented a significantly higher frequency of symptoms of fever, cough, fatigue, loss of taste, and loss of smell, as well as a higher rate of admission to the intensive care unit (ICU). Data analysis draws attention to the high number of cases of SARS without a causal diagnosis, the low access to ICU and orotracheal intubation (OTI), which might be explained by the demographic and regional inequalities in the access to healthcare.

The Effect of Being Pregnant during Respiratory Pandemics: A Comparison between 2009/10 Flu and 2020/21 COVID-19 Pandemic in Brazil.

Pregnant women undergo physiological changes that make them a challenging group of patients during pandemic respiratory diseases, as previously found during H1N1 2009 pandemic and recently ratified in COVID-19 pandemic. We conducted a retrospective cohort analysis on 5888 hospitalized women for H1N1 flu pandemic (2190 pregnant and 3698 non-pregnant) and 64,515 hospitalized women for COVID-19 pandemic (5151 pregnant and 59,364 non-pregnant), from the Brazilian national database, to compare demographic profile, clinical aspects, and mortality in childbearing aged women during both pandemics. Additionally, the effect of being pregnant was compared between both pandemics. In both pandemics, pregnant women were younger than non-pregnant women. Overall, pregnant women had lower frequencies of comorbidities and were less symptomatic. Among hospitalized women, pregnant women presented lower mortality rates than non-pregnant women (9.7% vs. 12.6%, p = 0.002 in the H1N1 pandemic and 9.7% vs. 17.4%, p < 0.001 in the COVID-19 pandemic) and this difference was statistically more pronounced in the COVID-19 pandemic, even after balancing pregnant and non-pregnant groups regarding age and chronic diseases.

Virulence modulation of Candida albicans biofilms by metal ions commonly released from orthodontic devices.

The installation of metal devices leads to an increase in the salivary concentration of metal ions and in the growth of salivary Candida spp. However, the relationship between released metal ions and Candida virulence has not been previously examined. The objective of this study was to evaluate whether metal ions affect fungal virulence. We prepared culture media containing Ni(2+), Fe(3+), Cr(3+), Co(2+) or a mixture of these metal ions at concentrations similar to those released in saliva of orthodontic patients. Biofilms of Candida albicans SC5314 were grown for 72 h and their biomasses were determined. The supernatants were analyzed for secretory aspartyl protease (SAP) and hemolysin activities. To verify changes in virulence following treatment with metals, proteolytic and hemolytic activities were converted into specific activities. The results revealed that all ions, except Co(2+), caused increases in biofilm biomass. In addition, Ni(2+) caused an increase in SAP activity and Fe(3+) reduced hemolytic activity. However, the SAP and hemolysin activities in the presence of the mixture of ions did not differ from those of control. These results indicate that metal ions released during the degradation of orthodontic appliances can modulate virulence factors in C. albicans biofilms.

Metabolic Control of Patients with Phenylketonuria in a Portuguese Metabolic Centre Comparing Three Different Recommendations.

Blood phenylalanine (Phe) is used as the primary marker to evaluate metabolic control. Our study aimed to describe the metabolic control of patients with phenylketonuria (PKU) comparing three different treatment recommendations (European guidelines/US guidelines/Portuguese consensus). This was a retrospective, observational, single centre study in patients with PKU collecting data on blood Phe levels from 2017. Nutritional intake data and sapropterin (BH4) prescription were collected at the last appointment of 2017. The final sample studied included 87 patients (48% females) [13 hyperphenylalaninemia; 47 mild PKU; 27 classical PKU] with a median age of 18 y (range: 1-36 y). The median number of blood Phe measurements for patients was 21 (range: 6-89). In patients aged < 12 y, the median blood Phe level was 300 µmol/L (range 168-480) and 474 µmol/L (range 156-1194) for patients ≥ 12 y. Overall, a median of 83% of blood Phe levels were within the European PKU guidelines target range. In patients aged ≥ 12 years, there was a higher median % of blood Phe levels within the European PKU guidelines target range (≥12 y: 84% vs. <12 y: 56%). In children < 12 y with classical PKU (n = 2), only 34% of blood Phe levels were within target range for all 3 guidelines and 49% with mild PKU (n = 11). Girls had better control than boys (89% vs. 66% median Phe levels within European Guidelines). Although it is clear that 50% or more patients were unable to achieve acceptable metabolic control on current treatment options, a globally agreed upper Phe target associated with optimal outcomes for age groups is necessary. More studies need to examine how clinics with dissimilar resources, different therapeutic Phe targets and frequency of monitoring relate to metabolic control.

Implementing a Transition Program from Paediatric to Adult Services in Phenylketonuria: Results After Two Years of Follow-Up with an Adult Team.

We aimed to report the implementation of a phenylketonuria (PKU) transition program and study the effects of follow-up with an adult team on metabolic control, adherence, and loss of follow-up. Fifty-five PKU patients were analysed in the study periods (SP): 2 years before (SP1) and after the beginning of adult care (SP2). Retrospective data on metabolic control and number of clinic appointments were collected for each SP, and protein intakes were analysed. In SP2, three patients (6%) were lost to follow-up. There was a small but statistically significant increase in median number of annual blood spots from SP1 to SP2: 11 (7-15) vs. 14 (7-20); p = 0.002. Mean ± SD of median blood Phe remained stable (525 ± 248 µmol/L vs. 552 ± 225 µmol/L; p = 0.100); median % of blood Phe < 480 µmol/L decreased (51 (4-96)% vs. 37 (5-85)%; p = 0.041) and median number of clinic appointments increased from SP1 to SP2: (5 (4-6) vs. 11 (8-13); p < 0.001). No significant differences were found regarding any parameter of protein intake. Our results suggest that the implementation of an adult service was successful as impact on metabolic control was limited and attendance remained high. Continuous dietetic care likely contributed to these results by keeping patients in follow-up and committed to treatment.

[Community Therapy and psychic distress in the family system: an approach based on the new paradigm of science]. / Terapia Comunitária e sofrimento psíquico no sistema familiar: um enfoque baseado no novo paradigma da ciência.

The scope of this study was to analyze the behavior patterns adopted by family members dealing with individuals in psychological distress within the scope of the relationships established in the family system, after participating in Community Therapy sessions. It was based on research with a qualitative approach using the research-action-intervention method. The intervention used centered around twelve Community Therapy sessions. Ten family members of people in psychological distress monitored by a Psychosocial Care Center took part in the study. Field notebook records, videos obtained with the tapes of the Community Therapy sessions, the interview material and Community Therapy itself were used as the empirical material production technique. The study revealed that the family members' understanding of psychological distress could be transformed as they attributed new meanings and performed fundamental actions to balanced living in the family. Community Therapy was seen as a practice that facilitated changes in the lives of its participants, as it contributed to generate social and individual interactions that stimulated family members to rethink concepts and to create spaces of coexistence and human and respectful care that permeated the family unit.

O presente estudo buscou analisar os comportamentos adotados por familiares que lidam com entes em sofrimento psíquico, no âmbito das relações estabelecidas no sistema familiar, depois de participarem de rodas de Terapia Comunitária. Pesquisa de abordagem qualitativa que utilizou o método da pesquisa-ação-intervenção. A intervenção utilizada foram doze rodas de Terapia Comunitária. Fizeram parte do estudo dez familiares de pessoas em sofrimento psíquico acompanhadas em um Centro de Atenção Psicossocial. Como técnicas de produção do material empírico foram utilizados os registros do caderno de campo, os vídeos obtidos com a gravação das rodas de Terapia Comunitária, o material das entrevistas e a própria Terapia Comunitária. O estudo revelou que a compreensão dos familiares acerca do sofrimento psíquico pôde ser transformada na medida que atribuíam novos significados e realizavam ações fundamentais ao convívio equilibrado na família. A Terapia Comunitária pôde ser elucidada como uma prática que facilitou mudanças na vida de seus participantes, pois contribuiu para gerar interações sociais e individuais que estimularam os familiares a repensarem conceitos e a criarem espaços de convivência e cuidado humanos e respeitosos que se difundiram para o sistema familiar.

Continuous use of glycomacropeptide in the nutritional management of patients with phenylketonuria: a clinical perspective.

BACKGROUND: In phenylketonuria (PKU), modified casein glycomacropeptide supplements (CGMP-AA) are used as an alternative to the traditional phenylalanine (Phe)-free L-amino acid supplements (L-AA). However, studies focusing on the long-term nutritional status of CGMP-AA are lacking. This retrospective study evaluated the long-term impact of CGMP-AA over a mean of 29 months in 11 patients with a mean age at CGMP-AA onset of 28 years (range 15-43) [8 females; 2 hyperphenylalaninaemia (HPA), 3 mild PKU, 3 classical PKU and 3 late-diagnosed]. Outcome measures included metabolic control, anthropometry, body composition and biochemical parameters. RESULTS: CGMP-AA, providing 66% of protein equivalent intake from protein substitute, was associated with no significant change in blood Phe with CGMP-AA compared with baseline (562 ± 289 µmol/L vs 628 ± 317 µmol/L; p = 0.065). In contrast, blood tyrosine significantly increased on CGMP-AA (52.0 ± 19.2 µmol/L vs 61.4 ± 23.8 µmol/L; p = 0.027). CONCLUSIONS: Biochemical nutritional markers remained unchanged which is an encouraging finding in adults with PKU, many of whom are unable to maintain full adherence with nutritionally fortified protein substitutes. Longitudinal, prospective studies with larger sample sizes are necessary to fully understand the metabolic impact of using CGMP-AA in PKU.

Over Restriction of Dietary Protein Allowance: The Importance of Ongoing Reassessment of Natural Protein Tolerance in Phenylketonuria.

Phenylalanine (Phe) tolerance is highly variable in phenylketonuria (PKU) and rarely described in patients aged ≥12 years. Patients ≥12 years of age with PKU were systematically challenged with additional natural protein (NP) if blood Phe levels remained below 480 µmol/L (i.e., upper target blood Phe level for patients aged ≥12 years using Portuguese PKU guidelines). In PKU patients, NP tolerance was calculated at baseline and a median of 6 months after systematic challenge with NP whilst patients were maintaining a blood Phe ≤480 µmol/L. Anthropometry was assessed at both times. Routine blood Phe levels were collected. We studied 40 well-controlled PKU patients (10 hyperphenylalaninemia (HPA), 23 mild and 7 classic PKU), on a low-Phe diet with a mean age of 17 years (12-29 years). Median daily NP intake significantly increased between assessments (35 vs. 40 g/day, p = 0.01). Twenty-six patients (65%) were able to increase their median NP intake by a median 12 g/day (2-42 g)/day and still maintain blood Phe within target range. Out of the previous 26 patients, 20 (77%) (8 HPA, 11 mild and 1 classical PKU) increased NP from animal sources (e.g. dairy products, fish and meat) and 6 patients (23%) (3 mild and 3 classical PKU) from plant foods (bread, pasta, potatoes). Median protein equivalent intake from Phe-free/low-Phe protein substitute decreased (0.82 vs. 0.75 g/kg, p = 0.01), while median blood Phe levels remained unchanged (279 vs. 288 µmol/L, p = 0.06). Almost two-thirds of patients with PKU tolerated additional NP when challenged and still maintained blood Phe within the national target range. This suggests that some patients with PKU treated by a low-Phe diet only may over restrict their NP intake. In order to minimise the burden of treatment and optimise NP intake, it is important to challenge with additional NP at periodic intervals.

The use of a custom-made mouthguard stabilizes the electromyographic activity of the masticatory muscles among Karate-Do athletes.

AIMS: To analyze and compare the electromyographic activity of the temporal (anterior portion) and masseter muscles among Karate-Do athletes before and after training, with and without the use of a mouthguard. METHODS: Twenty athletes (14 males and 6 females) with a mean age of 23.7 ± 7.5 years participated. They had surface electromyography recordings taken of their bilateral temporal and masseter muscles before and after training under the following conditions: no mouthguard, with a ready-made mouthguard, and with a custom-made mouthguard. Activity was examined at mandibular rest, while clenching, and at maximum voluntary contraction. The data were normalized using the mean maximum voluntary contraction. RESULTS: The right (p = 0.005) and left (p = 0.015) temporal muscles showed significantly lower electromyographic activity with a custom-made mouthguard compared with no mouthguard after training while clenching. The electromyographic activity of the temporal and masseter muscles did not show significant differences when tested at mandibular rest and while clenching before or after training with a custom-made mouthguard (p > 0.05). CONCLUSION: The use of a custom-made mouthguard preserved participants' electromyographic profiles before and after training; thus, they allow for stable muscle activity during the training of Karate-Do athletes.

Antimicrobial Activity of Two Garlic Species (Allium Sativum and A. Tuberosum) Against Staphylococci Infection. In Vivo Study in Rats.

Purpose: This study observed the effect of garlic extracts and amoxicillin against an induced staphylococcal infection model. MIC and MBC were also obtained for aqueous extracts of Allium sativum (Asa) and Allium tuberosum (Atu) against Staphylococcus aureus penicillin-sensitive (PSSA - ATCC 25923) and MRSA (ATCC 33592). Methods: Granulation tissues were induced in the back of 205 rats. After 14 days, 0.5 mL of 108 CFU/mL of PSSA or MRSA were injected inside tissues. After 24h, animals were divided: G1 (Control) - 0.5 mL of NaCl 0.9%; G2 - Asa 100 mg/kg or 400mg/kg; G3 - Atu 100 mg/kg or 400 mg/kg; G4 - amoxicillin suspension 50 mg/kg, considering PSSA infection; and G5 (Control) - 0.5 mL of NaCl 0.9%; G6 - Asa 400mg/kg; G7 - amoxicillin 50 mg/kg; and G8 - Asa 400 mg/kg + amoxicillin 50 mg/kg for MRSA. All treatments were administered P.O. every 6h. Animals were killed at 0, 6, 12 and 24h. Samples were spread on salt-mannitol agar. Colonies were counted after 18 h at 37 °C. Atu was not able to inhibit or kill PSSA and MRSA. Considering Asa, MIC and MBC against PSSA were 2 mg/mL and 4 mg/mL, respectively; and 16 mg/mL and 64 mg/mL against MRSA. Results: No effect was observed in vivo for control, Asa 100 mg/kg and Atu 100 mg/kg, while amoxicillin, Atu 400 mg/kg and Asa 400 mg/kg decreased PSSA counts in all-time points. No effect of any group against MRSA was observed at any time. Conclusion: Thus, A. sativum and A. tuberosum were able to reduce PSSA infection, but not MRSA infection.