Pathogenic Homocystinuria-Associated T236N Mutation Dramatically Alters the Biochemical Properties of Cystathionine Beta-Synthase Protein.

BACKGROUND: Cystathione beta-synthase (CBS) T236N is a novel mutation associated with pyridoxine non-responsiveness, which presents a significant difficulty in the medical treatment of homocystinuria. Reported severe phenotypes in homocystinuria patients highlight the urgent requirement to comprehend the molecular mechanisms underlying mutation pathogenicity for the advancement of the disease. METHODOLOGY: In this study, we used a multidisciplinary approach to investigate the molecular properties of bacterially expressed and purified recombinant CBST236N protein, which we directly compared to those of the wild-type (CBSWT) protein. RESULTS: Our data revealed a profound impact of the p.T236N mutation on CBS enzymatic activity, with a dramatic reduction of ~96% compared to the CBSWT protein. Circular dichroism (CD) experiments indicated that the p.T236N mutation did not significantly alter the secondary structure of the protein. However, CD spectra unveiled distinct differences in the thermal stability of CBSWT and CBST236N mutant protein species. In addition, chemical denaturation experiments further highlighted that the CBSWT protein exhibited greater thermodynamic stability than the CBST236N mutant, suggesting a destabilizing effect of this mutation. CONCLUSIONS: Our findings provide an explanation of the pathogenicity of the p.T236N mutation, shedding light on its role in severe homocystinuria phenotypes. This study contributes to a deeper understanding of CBS deficiency and may improve the development of targeted therapeutic strategies for affected individuals.

Arrhythmia-Associated Calmodulin E105A Mutation Alters the Binding Affinity of CaM to a Ryanodine Receptor 2 CaM-Binding Pocket.

Calmodulin (CaM) is a small, multifunctional calcium (Ca2+)-binding sensor that binds and regulates the open probability of cardiac ryanodine receptor 2 (RyR2) at both low and high cytosolic Ca2+ concentrations. Recent isothermal titration calorimetry (ITC) studies of a number of peptides that correspond to different regions of human RyR2 showed that two regions of human RyR2 (3584-3602aa and 4255-4271aa) bind with high affinity to CaM, suggesting that these two regions might contribute to a putative RyR2 intra-subunit CaM-binding pocket. Moreover, a previously characterized de novo long QT syndrome (LQTS)-associated missense CaM mutation (E105A) which was identified in a 6-year-old boy, who experienced an aborted first episode of cardiac arrest revealed that this mutation dysregulates normal cardiac function in zebrafish by a complex mechanism that involves alterations in both CaM-Ca2+ and CaM-RyR2 interactions. Herein, to gain further insight into how the CaM E105A mutation leads to severe cardiac arrhythmia, we generated large quantities of recombinant CaMWT and CaME105A proteins. We then performed ITC experiments to investigate and compare the interactions of CaMWT and CaME105A mutant protein with two synthetic peptides that correspond to the two aforementioned human RyR2 regions, which we have proposed to contribute to the RyR2 CaM-binding pocket. Our data reveal that the E105A mutation has a significant negative effect on the interaction of CaM with both RyR2 regions in the presence and absence of Ca2+, highlighting the potential contribution of these two human RyR2 regions to an RyR2 CaM-binding pocket, which may be essential for physiological CaM/RyR2 association and thus channel regulation.

The Relationship between Changes in MYBPC3 Single-Nucleotide Polymorphism-Associated Metabolites and Elite Athletes' Adaptive Cardiac Function.

Athletic performance is a multifactorial trait influenced by a complex interaction of environmental and genetic factors. Over the last decades, understanding and improving elite athletes' endurance and performance has become a real challenge for scientists. Significant tools include but are not limited to the development of molecular methods for talent identification, personalized exercise training, dietary requirements, prevention of exercise-related diseases, as well as the recognition of the structure and function of the genome in elite athletes. Investigating the genetic markers and phenotypes has become critical for elite endurance surveillance. The identification of genetic variants contributing to a predisposition for excellence in certain types of athletic activities has been difficult despite the relatively high genetic inheritance of athlete status. Metabolomics can potentially represent a useful approach for gaining a thorough understanding of various physiological states and for clarifying disorders caused by strength-endurance physical exercise. Based on a previous GWAS study, this manuscript aims to discuss the association of specific single-nucleotide polymorphisms (SNPs) located in the MYBPC3 gene encoding for cardiac MyBP-C protein with endurance athlete status. MYBPC3 is linked to elite athlete heart remodeling during or after exercise, but it could also be linked to the phenotype of cardiac hypertrophy (HCM). To make the distinction between both phenotypes, specific metabolites that are influenced by variants in the MYBPC3 gene are analyzed in relation to elite athletic performance and HCM. These include theophylline, ursodeoxycholate, quinate, and decanoyl-carnitine. According to the analysis of effect size, theophylline, quinate, and decanoyl carnitine increase with endurance while decreasing with cardiovascular disease, whereas ursodeoxycholate increases with cardiovascular disease. In conclusion, and based on our metabolomics data, the specific effects on athletic performance for each MYBPC3 SNP-associated metabolite are discussed.

Optimizing the Functional Properties of Starch-Based Biodegradable Films.

A definitive screening design was used in order to evaluate the effects of starch, glycerol and montmorillonite (MMT) concentrations, as well as the drying temperature, drying tray type and starch species, on packaging film's functional properties. Optimization showed that in order to obtain films with the minimum possible thickness, the maximum elongation at break, the maximum tensile strength, as well as reduced water vapor permeability and low opacity, a combination of factors should be used as follows: 5.5% wt starch concentration, 30% wt glycerol concentration on a dry starch basis, 10.5% wt MMT concentration on a dry starch basis, 45 °C drying temperature, chickpea as the starch species and plexiglass as the drying tray type. Based on these results, starch films were prepared, and fresh minced meat was stored in them for 3 days. It was shown that the incorporation of MMT at 10.5% wt on a dry starch basis in the packaging films led to a decreased mesophilic and psychrotrophic bacteria growth factor compared to commercial packaging. When assessed for their biodegradability, the starch films disintegrated after 10 days of thermophilic incubation under simulated composting conditions. Finally, to prove their handling capability during industrial production, the starch films were rewound in a paper cylinder using an industrial-scale rewinding machine.

Navigating paediatric virology through the COVID­19 era (Review).

The present review article presents the key messages of the 8th Workshop on Paediatric Virology organised virtually by the Institute of Paediatric Virology based on the island of Euboea in Greece. The major topics covered during the workshop were the following: i) New advances in antiviral agents and vaccines against cytomegalovirus; ii) hantavirus nephropathy in children; iii) human rhinovirus infections in children requiring paediatric intensive care; iv) complications and management of human adenovirus infections; v) challenges of post­coronavirus disease 2019 (COVID­19) syndrome in children and adolescents; and vi) foetal magnetic resonance imaging in viral infections involving the central nervous system. The COVID­19 era requires a more intensive, strategic, global scientific effort in the clinic and in the laboratory, focusing on the diagnosis, management and prevention of viral infections in neonates and children.

Exploring medical terminology inexpediencies: Tripledemic vs. triple epidemic.

Accurate and consistent medical terminology has a fundamental value in medicine. It enables medical students to understand the meaning of each term, medical physicians to communicate with each other, and it also enables science to adopt a logical language of high-level understanding and scientific regularity. Medical terminology inexpediencies caused by the adoption of etymologically illogical or linguistically false terms lead to misunderstanding and confusion among clinicians. The medical terms epidemic and pandemic are as old as Hippocrates and Sophocles, respectively. The present article evaluates the new medical terms tripledemic and triple epidemic, which were introduced during the recent COVID-19 pandemic.

Long-term effectiveness of non-surgical open-bite treatment: a systematic review and meta-analysis.

BACKGROUND: The etiology of open bite is complex, involving various genetic or environmental factors. Several treatment alternatives have been suggested for the correction of open bite, yet their long-term effectiveness remains controversial. OBJECTIVE: To assess the long-term effectiveness of open-bite treatment in treated with non-surgical approaches versus untreated patients, through lateral cephalometric radiographs. SEARCH METHODS: Unrestricted search of 16 electronic databases and manual searches up to November 2022. SELECTION CRITERIA: Randomized or non-randomized controlled trials reporting on the long-term effects of open-bite treatment through angular lateral cephalometric variables. DATA COLLECTION AND ANALYSIS: Only angular variables on lateral cephalometric radiographs were considered as primary outcomes. For each outcome, the mean differences and 95% confidence intervals were calculated using the random-effects model to consider existing heterogeneity. The revised Cochrane risk-of-bias tool (R.o.B. 2.0) and the risk-of-bias tool for non-randomized studies for interventions (ROBINS-I) were utilized for the randomized and non-randomized trials, respectively. RESULTS: From the initially identified 26,527 hits, only 6 studies (1 randomized and 5 retrospective controlled trials) were finally included in this systematic review reporting on 244 open-bite individuals (134 patients and 110 untreated controls), while five of them were included in the meta-analyses, assessing either the interval ranging from treatment start to post-retention (T3-T1) or from end of treatment to post-retention period (T3-T2). Regarding the vertical plane, for the T3-T2 interval, no significant differences were found for the assessed skeletal measurements, indicating a relative stability of the treatment results. Similarly, with regard to the T3-T1 interval, no significant differences could be identified for the examined skeletal variables, implying that the produced effects are rather minimal and that the correction of the open bite was performed mainly through dentoalveolar rather than skeletal changes. Further, no significant changes could be identified regarding the inclination of the upper and lower incisors. Only the nasolabial angle was significantly reduced in the treated patients in the long term. CONCLUSIONS: According to existing evidence, the influence of non-surgical treatment of open bite on the skeletal tissues and the inclination of the incisors is rather minimal in the long term, while only the nasolabial angle was significantly reduced.

Vaccination programs for children aged up to 18 years in Europe, 2020.

Although all European countries have vaccination policies for children, there are no comprehensive studies of pediatric vaccination programs in Europe. We studied vaccination programs for children in Europe. Vaccinations against diphtheria, tetanus, pertussis, poliomyelitis, Haemophilus influenzae type b, hepatitis B, measles, mumps, rubella, and influenza existed in 42 countries, against human papilloma virus in 41 countries, and against pneumococcus in 40 countries. In addition, the following vaccinations existed: against tuberculosis (35 countries), hepatitis A (33), meningococcus A, C, W, Y (30), rotavirus and varicella (28 countries each), meningococcus B (24), tick-born encephalitis (22), and meningococcus C (16). Mandatory vaccinations are implemented in 21 countries, mainly against diphtheria, tetanus, pertussis, poliomyelitis, H. influenzae type b, hepatitis B, measles, mumps, rubella, tuberculosis, and pneumococcus. There are significant differences among pediatric vaccination programs in Europe regarding number, schedules, indications, and regulatory frame (recommended or mandatory vaccinations). A consensus-based vaccination program for all children is needed.

Adherence to Recommendations for Βacillus Calmette-Guérin Vaccination of High-risk Neonates in Greece.

In 2016 a Βacillus Calmette-Guérin vaccination policy targeting high-risk neonates for tuberculosis before discharge from maternity hospital was adopted in Greece. Vaccination rates were 38.2% in 2019 and 24.7% in 2020. Vaccination coverage varied by risk group (higher for neonates in close contact with an active noncompliant or multidrug-resistant tuberculosis case and lower for Roma and immigrant neonates).

Effectiveness of maternal vaccination with quadrivalent inactivated influenza vaccine in pregnant women and their infants in 2019-2020.

OBJECTIVES: Influenza is associated with an increased risk for serious illness, hospitalization, and death in pregnant women and young infants. Our aim was to estimate the effectiveness of a quadrivalent inactivated influenza vaccine (QIV) in pregnant women and their infants during 2019-2020. METHODS: A QIV vaccine was offered to pregnant women followed in a maternity hospital. Women were contacted weekly during the influenza season and asked about symptoms. Polymerase chain reaction testing in pharyngeal samples was offered to pregnant women and infants with influenza-like illness. A Bayesian beta-binomial model was used. RESULTS: We studied 636 pregnant women (406 vaccinated and 230 unvaccinated) and 474 infants (281 of mothers vaccinated in pregnancy and 193 of unvaccinated mothers). Using a Bayesian beta-binomial model, it was estimated that influenza vaccination of pregnant women reduced their logit to develop laboratory-confirmed influenza by -4.2 (95% CI -3,7 - 4,7) and the logit of their infants to develop laboratory-confirmed influenza by -4.2 (95% CI -3.6, -4.9). The QIV effectiveness against laboratory-confirmed influenza was 43.5% in pregnant women and 31.4% in infants. CONCLUSION: Maternal influenza vaccination with QIV in pregnancy reduced the odds of pregnant women and their infants to develop influenza. CLINICAL TRIAL REGISTRATION: www.clinicaltrials.gov identifier is NCT04723771.

Insights in paediatric virology during the COVID-19 era (Review).

The present article provides an overview of the key messages of the topics discussed at the '7th Workshop on Paediatric Virology', which was organised virtually on December 20, 2021 by the Institute of Paediatric Virology, located on the Island of Euboea in Greece. The workshop's plenary lectures were on: i) viral pandemics and epidemics in the ancient Mediterranean; ii) the impact of obesity on the outcome of viral infections in children and adolescents; and iii) COVID-19 and artificial intelligence. Despite the scarcity of evidence from fossils and remnants, viruses have been recognised as significant causes of several epidemics in the ancient Mediterranean. Paediatric obesity, a modifiable critical health risk factor, has been shown to impact on the development, progression and severity of viral infections. Thus, the prevention of paediatric obesity should be included in formulating public health policies and decision-making strategies against emerging global viral threats. During the current COVID-19 pandemic, artificial intelligence has been used to facilitate the identification, monitoring and prevention of SARS-CoV-2. In the future, it will play a fundamental role in the surveillance of epidemic-prone infectious diseases, in the repurposing of older therapies and in the design of novel therapeutic agents against viral infections. The collaboration between different medical specialties and other diverse scientific fields, including archaeology, history, epidemiology, nutritional technologies, mathematics, computer technology, engineering, medical law and ethics is essential for the successful management of paediatric viral infections. The current COVID-19 pandemic has underscored this need, which should be further encouraged in modern medical education.

Vaccination programs for pregnant women in Europe, 2021.

Vaccination during pregnancy is increasingly adopted worldwide in order to protect the mother and her offspring. We studied the current vaccination programs specifically for pregnant women in 42 European countries. Vaccination programs for pregnant women are in place in 37 countries, as follows: influenza (36 countries), pertussis (28), hepatitis B (12), tetanus (10), pneumococcal disease (10), meningococcal disease (10), rabies (8), tick-borne encephalitis (6), hepatitis A (5), poliomyelitis (4), diphtheria (3), Haemophilus influenzae (2), and human papilloma virus (1). Recommendations for vaccination against influenza and pertussis concern almost exclusively pregnant women regardless of high-risk conditions, however differences between vaccination recommendations are noted in terms of timing. Vaccinations against hepatitis B, hepatitis A, pneumococcal disease, meningococcal disease, poliomyelitis, H. influenzae, rabies, and tick-born encephalitis mainly concern pregnant women at high-risk for exposure or serious illness and post-exposure vaccinations. Overall, five European countries have no vaccination recommendations specifically for pregnant women. In conclusion, there are significant differences in vaccination programs for pregnant women in Europe. Vaccination programs for pregnant women should expand in order to protect maternal and infant health. A consensus-based vaccination program is needed.

Post-COVID Syndrome: Incidence, Clinical Spectrum, and Challenges for Primary Healthcare Professionals.

Post-COVID syndrome also known as long COVID refers to symptoms persisting for more than three weeks after the diagnosis of COVID-19. We reviewed the current evidence on post-COVID syndrome, focusing on its clinical manifestations and addressing the challenges for its management in primary healthcare. The incidence of post-COVID syndrome is estimated at 10-35%, while for hospitalized patients it may reach 85%. Fatigue is the most common symptom reported in 17.5-72% of post-COVID cases, followed by residual dyspnea with an incidence ranging from 10-40%. Mental problems, chest pain, and olfactory and gustatory dysfunction may affect up to 26, 22 and 11% of patients, respectively. More than one third of patients with post-COVID syndrome have pre-existing comorbidities, hypertension and diabetes mellitus being the most common. Beyond the prolonged duration of symptoms, the scarce published data indicate that most patients with post-COVID syndrome have a good prognosis with no further complications or fatal outcomes reported. Given the clinical spectrum of patients with post-COVID syndrome, most of them will be managed by primary healthcare professionals, in conjunction with pre-existing or new co-morbidities, which, in turn, may increase the burden of COVID-19 on primary healthcare. In conclusion approximately 10% of patients with COVID-19 may have symptoms persisting beyond three weeks, fulfilling the criteria of post-COVID syndrome. Primary healthcare professionals have a key role in the management of patients with post-COVID syndrome. Research is needed to elucidate the pathogenesis, clinical spectrum, and prognosis of post-COVID syndrome.

Determinants of intention to get vaccinated against COVID-19 among healthcare personnel in hospitals in Greece.

BACKGROUND: To investigate intention rates to get vaccinated against COVID-19 among healthcare personnel (HCP) in Greece. METHODS: Cross-sectional survey. RESULTS: The response rate was 14.5%. Of 1521 HCP with a known profession, 607 (39.9%) were nursing personnel, 480 (31.6%) physicians, 171 (11.2%) paramedical personnel, 72 (4.7%) supportive personnel, and 191 (12.6%) administrative personnel. Overall, 803 of 1571 HCP (51.1%) stated their intention to get vaccinated while 768 (48.9%) stated their intention to decline vaccination. Most HCP (71.3%) who reported intent to get vaccinated noted contributing to the control of the pandemic and protecting their families and themselves as their reasons, while the most common reason for reporting intent to decline vaccination was inadequate information about the vaccines (74.9%), followed by concerns about vaccine safety (36.2%). Logistic regression analysis revealed that the probability of intending to get vaccinated increased with male gender, being a physician, history of complete vaccination against hepatitis B, history of vaccination against pandemic A (H1N1) in 2009-2010, belief that COVID-19 vaccination should be mandatory for HCP, and increased confidence in vaccines in general during the COVID-19 pandemic. The following factors were associated with a lower intention to get vaccinated: no vaccination against influenza the past season, no intention to get vaccinated against influenza in 2020-2021, and no intention to recommend COVID-19 vaccination to high-risk patients. CONCLUSION: There is an urgent need to built safety perception towards COVID-19 vaccines and raise vaccine uptake rates by HCP, and thus to protect the healthcare workforce and the healthcare services.

COVID-19 and paediatric challenges: An interview with Professor of Paediatrics Vana Papaevangelou (University of Athens School of Medicine).

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a novel coronavirus, which causes coronavirus disease 2019 (COVID-19) and affects children less frequently than adults. According to Professor Vana Papaevangelou, Professor of Paediatrics at the University of Athens School of Medicine, children comprise only 2-6% of COVID-19 cases, worldwide, and they are not considered as super-spreaders of this infection. SARS-CoV-2 is transmitted through droplets, fomites, aerosol and fecal-oral route, while there is no strong evidence as yet, supporting transplacental transmission. Professor Papaevangelou highlights the epidemiological differences between seasonal influenza and COVID-19 and accepts that school closure had no direct impact since children are not the main transmitters of SARS-CoV-2. On the other hand, social distancing clearly limited the transmission of SARS-CoV-2, while quarantine seemed necessary during the first wave of this pandemic. She refers to antivirals, as well as other therapeutic agents able to diminish the immune response producing multisystem inflammatory syndrome, which is associated with increased mortality, and she notes that these agents were rarely used in children with COVID-19, while in most cases supportive treatment sufficed. She finishes with the ongoing scientific efforts for the development of an effective and safe vaccine against SARS-CoV-2 indicating that so far the most promising vaccine developments include vaccines that use viral vectors.