RESUMEN
Highly conserved transport protein particle (TRAPP) complexes regulate subcellular trafficking pathways. Accurate protein trafficking has been increasingly recognized to be critically important for normal development, particularly in the nervous system. Variants in most TRAPP complex subunits have been found to lead to neurodevelopmental disorders with diverse but overlapping phenotypes. We expand on limited prior reports on TRAPPC6B with detailed clinical and neuroradiologic assessments, and studies on mechanisms of disease, and new types of variants. We describe 29 additional patients from 18 independent families with biallelic variants in TRAPPC6B. We identified seven homozygous nonsense (n = 12 patients) and eight canonical splice-site variants (n = 17 patients). In addition, we identified one patient with compound heterozygous splice-site/missense variants with a milder phenotype and one patient with homozygous missense variants. Patients displayed non-progressive microcephaly, global developmental delay/intellectual disability, epilepsy and absent expressive language. Movement disorders including stereotypies, spasticity and dystonia were also observed. Brain imaging revealed reductions in cortex, cerebellum and corpus callosum size with frequent white matter hyperintensity. Volumetric measurements indicated globally diminished volume rather than specific regional losses. We identified a reduced rate of trafficking into the Golgi apparatus and Golgi fragmentation in patient-derived fibroblasts that was rescued by wild-type TRAPPC6B. Molecular studies revealed a weakened interaction between mutant TRAPPC6B (c.454C>T, p.Q152*) and its TRAPP binding partner TRAPPC3. Patient-derived fibroblasts from the TRAPPC6B (c.454C>T, p.Q152*) variant displayed reduced levels of TRAPPC6B as well as other TRAPP II complex-specific members (TRAPPC9 and TRAPPC10). Interestingly, the levels of the TRAPPC6B homologue TRAPPC6A were found to be elevated. Moreover, co-immunoprecipitation experiments showed that TRAPPC6A co-precipitates equally with TRAPP II and TRAPP III, while TRAPPC6B co-precipitates significantly more with TRAPP II, suggesting enrichment of the protein in the TRAPP II complex. This implies that variants in TRAPPC6B may preferentially affect TRAPP II functions compared to TRAPP III functions. Finally, we assessed phenotypes in a Drosophila TRAPPC6B-deficiency model. Neuronal TRAPPC6B knockdown impaired locomotion and led to wing posture defects, supporting a role for TRAPPC6B in neuromotor function. Our findings confirm the association of damaging biallelic TRAPPC6B variants with microcephaly, intellectual disability, language impairments, and epilepsy. A subset of patients also exhibited dystonia and/or spasticity with impaired ambulation. These features overlap with disorders arising from pathogenic variants in other TRAPP subunits, particularly components of the TRAPP II complex. These findings suggest that TRAPPC6B is essential for brain development and function, and TRAPP II complex activity may be particularly relevant for mediating this function.
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Distonía , Epilepsia , Discapacidad Intelectual , Microcefalia , Trastornos del Neurodesarrollo , Animales , Humanos , Microcefalia/genética , Discapacidad Intelectual/genética , Proteínas de Transporte Vesicular/genética , Trastornos del Neurodesarrollo/genética , Epilepsia/genéticaRESUMEN
Proline-5-carboxylate reductase 2, encoded by PYCR2 gene, is an enzyme that catalyzes the last step of proline synthesis from pyrroline-5-carboxylate synthetase to proline. PYCR2 gene defect causes hypomyelinating leukodystrophy 10. Up until now, to our knowledge around 38 patients with PYCR2 defect have been reported. Herein, we describe clinical, neuroradiological, biochemical findings, and metabolomic profiling of three new genetically related cases of PYCR2 defects from a large family. Cerebrospinal fluid (CSF) amino acid levels were measured and untargeted metabolomic profiling of plasma and CSF were conducted and evaluated together with the clinical findings in the patients. While plasma and CSF proline levels were found to be totally normal, untargeted metabolomic profiling revealed mild increases of glutamate, alpha-ketoglutarate, and l-glutamate semialdehyde and marked increases of inosine and xanthine. Our findings and all the previous reports suggest that proline auxotrophy is not the central disease mechanism. Untargeted metabolomics point to mild changes in proline pathway and also in purine/pyrimidine pathway.
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Enfermedades Desmielinizantes del Sistema Nervioso Central Hereditarias , Metabolómica , Prolina , Pirrolina Carboxilato Reductasas , Niño , Femenino , Humanos , Masculino , delta-1-Pirrolina-5-Carboxilato Reductasa , Ácido Glutámico/metabolismo , Enfermedades Desmielinizantes del Sistema Nervioso Central Hereditarias/genética , Enfermedades Desmielinizantes del Sistema Nervioso Central Hereditarias/patología , Enfermedades Desmielinizantes del Sistema Nervioso Central Hereditarias/diagnóstico , Ácidos Cetoglutáricos/metabolismo , Ácidos Cetoglutáricos/sangre , Imagen por Resonancia Magnética , Redes y Vías Metabólicas/genética , Metaboloma/genética , Metabolómica/métodos , Mutación/genética , Linaje , Prolina/líquido cefalorraquídeo , Purinas/metabolismo , Pirimidinas , Pirrolina Carboxilato Reductasas/genética , Pirrolina Carboxilato Reductasas/deficiencia , Xantina/sangre , LactanteRESUMEN
One significant constraint in the advancement of biosensors is the signal-to-noise ratio, which is adversely affected by the presence of interfering factors such as blood in the sample matrix. In the present investigation, a specific aptamer binding was chosen for its affinity, while exhibiting no binding affinity towards non-target bacterial cells. This selective binding property was leveraged to facilitate the production of magnetic microparticles decorated with aptamers. A novel assay was developed to effectively isolate S. pneumoniae from PBS or directly from blood samples using an aptamer with an affinity constant of 72.8 nM. The capture experiments demonstrated efficiencies up to 87% and 66% are achievable for isolating spiked S. pneumoniae in 1 mL PBS and blood samples, respectively.
Asunto(s)
Aptámeros de Nucleótidos , Dióxido de Silicio , Aptámeros de Nucleótidos/química , Dióxido de Silicio/química , Streptococcus pneumoniae/aislamiento & purificación , Streptococcus pneumoniae/química , Humanos , Técnicas Biosensibles/métodos , Nanopartículas de Magnetita/químicaRESUMEN
OBJECTIVES: Mechano-sensitive odontoblast cells, which sense mechanical loading and various stresses in the tooth structure, synthesize early signaling molecules such as prostaglandin E2 (PGE2) and nitric oxide (NO) as an adaptive response. It is thought that these synthesized molecules can be used for the diagnosis and treatment of periodontal and periapical diseases. The aim of this study was to investigate the relationship between the severity of apical periodontitis (AP) and chronic periodontitis (CP) and serum (s) TNF-α, IL-10, PGE2 and NO levels, as well as PGE2 and NO levels in gingival crevicular fluid (GCF) samples. MATERIALS & METHODS: A total of 185 subjects were divided into three categories: AP group (n = 85), CP group (n = 50) and healthy control group (n = 50). The AP group was divided into 3 subgroups according to abscess scoring (AS-PAI 1, 2 and 3) based on the periapical index. The CP group was divided into 4 subgroups according to the periodontitis staging system (PSS1, 2,3 and 4). After recording the demographic and clinical characteristics of all participants, serum (s) and gingival crevicular fluid (GCF) samples were taken. TNF-α, IL-10, PGE2 and NO levels were measured in these samples. RESULTS: Unlike serum measurements (sTNF-α, sIL-10, sNO and sPGE2), GCF-NO and GCF-PGE levels of the AP group were significantly higher than the control group in relation to abscess formation (54.4 ± 56.3 vs. 22.5 ± 12.6 µmol/mL, p < 0.001 and 100 ± 98 vs. 41 ± 28 ng/L, p < 0.001, respectively). Confirming this, the GCF-NO and GCF-PGE levels of the AS-PAI 1 group, in which abscesses have not yet formed, were found to be lower than those in AS-PAI 2 and 3, which are characterized by abscess formation [(16.7(3.7-117.8), 32.9(11.8-212.8) and 36.9(4.3-251.6) µmol/mL, p = 0,0131; 46.0(31.4-120.0), 69.6(40.3-424.2) and 74.4(32.1-471.0) ng/L, p = 0,0020, respectively]. Consistent with the increase in PSS, the levels of sTNF [29.8 (8.2-105.5) vs. 16.7(6.3-37.9) pg/mL, p < 0.001], sIL-10 [542(106-1326) vs. 190(69-411) pg/mL, p < 0.001], sNO [182.1(36.3-437) vs. 57.0(15.9-196) µmol/mL, p < 0.001], sPGE2 [344(82-1298) vs. 100(35-1178) ng/L, p < 0.001], GCF-NO [58.9 ± 33.6 vs. 22.5 ± 12.6 ng/L, p < 0.001] and GCF-PGE2 [ 99(37-365) vs. 30(13-119), p < 0.001] in the CP group were higher than the control group. Comparison ROC analysis revealed that the GCF-PGE2 test had the best diagnostic value for both AP and CP (sensitivity: 94.1 and 88.0; specificity: 64.0 and 78.0, respectively; p < 0.001). CONCLUSIONS: GCF-PE2 and GCF-NO have high diagnostic value in the determination of AP and CP, and can be selected as targets to guide treatment. In addition, the measurements of PGE2 and NO in GCF can be used as an important predictor of pulpal necrosis leading to abscess in patients with AP. CLINICAL RELEVANCE: In this article, it is reported that syntheses of early signaling molecules such as PGE2 and NO can be used for the diagnosis and treatment target of periapical and periodontal infections.
Asunto(s)
Periodontitis Crónica , Dinoprostona , Líquido del Surco Gingival , Interleucina-10 , Óxido Nítrico , Periodontitis Periapical , Factor de Necrosis Tumoral alfa , Humanos , Periodontitis Periapical/metabolismo , Masculino , Femenino , Periodontitis Crónica/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico/biosíntesis , Líquido del Surco Gingival/química , Adulto , Dinoprostona/metabolismo , Interleucina-10/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Persona de Mediana Edad , Ensayo de Inmunoadsorción Enzimática , Estudios de Casos y ControlesRESUMEN
Bacterial products, host immune cells and cytokines have been reported to play an important role in the pathogenesis of apical periodontitis (AP). This study aimed to determine the main bacterial species in the microbiota as gram positive and negative and to compare the relationship between matrix metalloproteinase (MMP)-9 and tumor necrosis factor (TNF)-α with controlled patient groups. 60 patients with AP and extraction indication were included in the study. 30 systemically healthy volunteers without AP were selected as the control group. After access cavity preparation, an initial microbiologic sample (S1) was taken from the root canal. After atraumatic extraction of the tooth, a second microbial sample (S2) was taken from the extraradicular region. After bacterial DNA extraction, 16S rRNA gene primer was designed for sequence analysis. Bacterial community profiling was made by Sanger sequencing of the PCR products. In addition, serum MMP-9 and TNF-α levels were measured from all patients. TNF-α levels of the AP group were higher than the control group, while MMP-9 levels were found to be lower (p = 0.0264 and p = 0.0146, respectively). There was no difference in the main bacterial species isolated from the samples taken from the intracanal and extraradicular region of the tooth with AP (p = 0.714). The main bacterial species in the intracanal region of the tooth with AP are similar to the main bacterial species in the extraradicular region. The pathophysiology of the tooth with AP is associated with low MMP-9 and high TNF-α, independent of the bacterial species in the intracanal and extraradicular regions.
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Metaloproteinasa 9 de la Matriz , Microbiota , Periodontitis Periapical , ARN Ribosómico 16S , Factor de Necrosis Tumoral alfa , Humanos , Periodontitis Periapical/microbiología , Masculino , Femenino , Adulto , Factor de Necrosis Tumoral alfa/sangre , ARN Ribosómico 16S/análisis , ADN Bacteriano/análisis , Estudios de Casos y Controles , Persona de Mediana Edad , Reacción en Cadena de la PolimerasaRESUMEN
Objective: Lower extremity ischemia-reperfusion injury (IRI) may occur with trauma-related vascular injury and various vascular diseases, during the use of a tourniquet, in temporary clamping of the aorta in aortic surgery, or following acute or bilateral acute femoral artery occlusion. Mitochondrial dysfunction and increased basal oxidative stress in diabetes may cause an increase in the effects of increased reactive oxygen species (ROS) and mitochondrial dysfunction due to IRI. It is of great importance to examine therapeutic approaches that can minimize the effects of IRI, especially for patient groups under chronic oxidative stress such as DM. Cerium oxide (CeO2) nanoparticles mimic antioxidant enzymes and act as a catalyst that scavenges ROS. In this study, it was aimed to investigate whether CeO2 has protective effects on skeletal muscles in lower extremity IRI in mice with streptozocin-induced diabetes. Methods: A total of 38 Swiss albino mice were divided into six groups as follows: control group (group C, n = 6), diabetes group (group D, n = 8), diabetes-CeO2 (group DCO, n = 8), diabetes-ischemia/reperfusion (group DIR, n = 8), and diabetes-ischemia/reperfusion-CeO2 (group DIRCO, n = 8). The DCO and DIRCO groups were given doses of CeO2 of 0.5 mg/kg intraperitoneally 30 min before the IR procedure. A 120 min ischemia-120 min reperfusion period with 100% O2 was performed. At the end of the reperfusion period, muscle tissues were removed for histopathological and biochemical examinations. Results: Total antioxidant status (TAS) levels were found to be significantly lower in group DIR compared with group D (p = 0.047 and p = 0.022, respectively). In group DIRCO, total oxidant status (TOS) levels were found to be significantly higher than in group DIR (p < 0.001). The oxidative stress index (OSI) was found to be significantly lower in group DIR compared with group DCO (p < 0.001). Paraoxanase (PON) enzyme activity was found to be significantly increased in group DIR compared with group DCO (p < 0.001). The disorganization and degeneration score for muscle cells, inflammatory cell infiltration score, and total injury score in group DIRCO were found to be significantly lower than in group DIR (p = 0.002, p = 0.034, and p = 0.001, respectively). Conclusions: Our results confirm that CeO2, with its antioxidative properties, reduces skeletal muscle damage in lower extremity IRI in diabetic mice.
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Cerio , Diabetes Mellitus Experimental , Músculo Esquelético , Estrés Oxidativo , Daño por Reperfusión , Animales , Cerio/farmacología , Cerio/uso terapéutico , Ratones , Músculo Esquelético/efectos de los fármacos , Diabetes Mellitus Experimental/complicaciones , Estrés Oxidativo/efectos de los fármacos , Masculino , Estreptozocina , Antioxidantes/farmacología , Antioxidantes/uso terapéutico , Modelos Animales de Enfermedad , Especies Reactivas de Oxígeno/metabolismoRESUMEN
OBJECTIVES: To distinguish whether idiopathic intracranial hypertension (IIH) is a condition predisposing to multiple sclerosis (MS) or an isolated disease, the current gene transcription factor Activator Protein-1 (AP-1) was evaluated with its potential to differentiate both diseases. BACKGROUND: The aim of this study was to investigate the use of AP-1 as biomarkers for the discrimination of IIH and MS. METHODS: AP-1, TNF-α, and IL-6 protein values in the CSF of the cases were evaluated by the ELISA method. The numerical measures of the groups and the ability of AP-1 to distinguish the groups were analyzed with the ROC curve. RESULTS: There was no difference between the groups in CSF TNF-α, IL-6, CSF, and serum biochemistry analyses. However, it was determined that the AP-1 concentration (pg/ml) was significantly higher in the IIH group, the sensitivity of AP-1 in separating those with IIH was 75%, and the specificity in separating those with MS was 60% in those with an AP-1 concentration of 606.5 and above. CONCLUSION: According to our results, the fact that CSF TNF-α and IL-6 values did not differ in IIH compared to MS revealed that IIH could not methodologically control MS, and AP-1 was a supportive parameter in differentiating both diseases (Tab. 2, Fig. 1, Ref. 31).
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Biomarcadores , Esclerosis Múltiple , Factor de Transcripción AP-1 , Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Biomarcadores/líquido cefalorraquídeo , Diagnóstico Diferencial , Interleucina-6/líquido cefalorraquídeo , Esclerosis Múltiple/líquido cefalorraquídeo , Esclerosis Múltiple/diagnóstico , Seudotumor Cerebral/líquido cefalorraquídeo , Seudotumor Cerebral/diagnóstico , Curva ROC , Sensibilidad y Especificidad , Factor de Transcripción AP-1/líquido cefalorraquídeo , Factor de Transcripción AP-1/metabolismo , Factor de Necrosis Tumoral alfa/líquido cefalorraquídeoRESUMEN
AIM: The aim was to compare SARS-CoV-2 IgG antibody levels in chronic hepatitis B patients and healthcare personnel selected as the control group and to determine factors such as age, gender, vaccine type, and number of vaccines that may affect the antibody levels. MATERIALS AND METHODS: 87 chronic hepatitis B (CHB) patients followed in Ankara Training and Research Hospital Infectious Diseases Clinic and Mamak State Hospital Infectious Diseases outpatient clinic and 89 healthcare personnel selected as the control group were included in the study.SARS-CoV-2 IgG antibody levels in the serum samples of patients and healthcare personnel who received the COVID-19 vaccine were studied with the ELISA method in the Microbiology Laboratory of Ankara Training and Research Hospital, using a commercial ELISA kit (Abbott, USA) in line with the recommendations of the manufacturer. In the study, SARS-CoV-2 IgG levels were compared in CHB patients and healthcare personnel. In addition, the relationship between SARS-CoV-2 antibody level, gender, average age, natural history of the disease, number of vaccinations, vaccine type (Coronavac TM vaccine alone, BNT162b2 vaccine alone or Coronavac TM and BNT162b2 vaccine (heterologous vaccination)), treatment duration of CHB was investigated. Statistical analyses were made in the SPSS program. A value of p≤ 0.05 was considered statistically significant. FINDINGS: A total of 167 people, including 87 CKD patients and 80 healthcare personnel as the control group, were included in the study. SARS-CoV-2 IgG antibody levels were detected above the cut-off level in the entire study group, regardless of the vaccine type. No difference was detected in SARS-CoV-2 IgG titers after COVID-19 vaccination between CHB patients and healthcare personnel. There was a statistically significant difference in SARS-CoV-2 IgG antibody levels among individuals participating in the study according to vaccine types. Compared to those who received Coronavac TM vaccine alone, the average SARS-CoV-2 IgG level was found to be statistically significantly higher in those who received BNT162b2 vaccine alone or heterologous vaccination with Coronavac TM + BNT162b2 vaccine. There was no difference between the groups in terms of age, gender, number of vaccinations, natural transmission of the disease, and duration of antiviral therapy in the CHD patient group. CONCLUSION: As a result, SARS-CoV-2 IgG antibody levels above the cut-off value were achieved with Coronavac TM and BNT162b2 vaccines in both CHD patients and healthy control groups. however, both CHD patients and healthcare personnel had higher antibody levels than those who received BNT162b2 alone or those who received heterologous vaccination had higher antibody levels than those with Coronavac TM alone. Therefore, if there are no contraindications, BNT162b2 vaccine may be preferred in CHB and health personnel (Tab. 2, Ref. 14).
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Anticuerpos Antivirales , Vacuna BNT162 , Vacunas contra la COVID-19 , COVID-19 , Hepatitis B Crónica , Inmunoglobulina G , SARS-CoV-2 , Humanos , Femenino , Masculino , Adulto , Persona de Mediana Edad , Vacunas contra la COVID-19/inmunología , COVID-19/prevención & control , COVID-19/inmunología , COVID-19/sangre , Inmunoglobulina G/sangre , Anticuerpos Antivirales/sangre , SARS-CoV-2/inmunología , Hepatitis B Crónica/inmunología , Hepatitis B Crónica/sangre , Vacuna BNT162/inmunología , Personal de Salud , Anciano , Adulto JovenRESUMEN
BACKGROUND: Old age, obesity, and certain chronic conditions are among the risk factors for severe COVID-19. More information is needed on whether inherited metabolic disorders (IMD) confer risk of more severe COVID-19. We aimed to establish COVID-19 severity and associated risk factors in patients with IMD currently followed at a single metabolic center. METHODS: Among all IMD patients followed at a single metabolic referral center who had at least one clinic visit since 2018, those with accessible medical records were reviewed for SARS-CoV-2 tests. COVID-19 severity was classified according to the WHO recommendations, and IMD as per the international classification of IMD. RESULTS: Among the 1841 patients with IMD, 248 (13.5%) had tested positive for COVID-19, 223 of whom gave consent for inclusion in the study (131 children and 92 adults). Phenylalanine hydroxylase (48.4%) and biotinidase (12.1%) deficiencies were the most common diagnoses, followed by mucopolysaccharidoses (7.2%). 38.1% had comorbidities, such as neurologic disabilities (22%) or obesity (9.4%). The majority of COVID-19 episodes were asymptomatic (16.1%) or mild (77.6%), but 6 patients (2.7%) each had moderate and severe COVID-19, and two (0.9%) had critical COVID-19, both of whom died. 3 patients had an acute metabolic decompensation during the infection. Two children developed multisystem inflammatory syndrome (MIS-C). Long COVID symptoms were present in 25.2%. Presence of comorbidities was significantly associated with more severe COVID-19 in adults with IMD (p < 0.01), but not in children (p = 0.45). Compared to other categories of IMD, complex molecule degradation disorders were significantly associated with more severe COVID-19 in children (p < 0.01); such a significant IMD category distinction was not found in adults. DISCUSSION: This is the largest study on COVID-19 in IMD patients relying on real-word data and objective definitions, and not on merely expert opinions or physician surveys. COVID-19 severity and long COVID incidence in IMD are probably similar to the general population, and the risk of acute metabolic decompensation is not likely to be greater than that in other acute infections. Disease category (complex molecule degradation) in children, and comorbidities in adults may be associated with COVID-19 severity in IMD. Additionally, the first documented accounts of COVID-19 in 27 different IMD are recorded. The high occurrence of MIS-C may be coincidental, but warrants further study.
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COVID-19 , Enfermedades Metabólicas , Adulto , Niño , Humanos , COVID-19/epidemiología , SARS-CoV-2 , Síndrome Post Agudo de COVID-19 , Factores de Riesgo , Gravedad del Paciente , Enfermedades Metabólicas/epidemiología , Obesidad/complicaciones , Obesidad/epidemiologíaRESUMEN
BACKGROUND: Phenylalanine (Phe)-restricted diet is associated with lower quality of life for patients with phenylketonuria (PKU), and a concern for caregivers of recently-diagnosed infants. Sapropterin is an oral drug used as an alternative or adjunct to dietary treatment. We have observed that some of the young infants initially managed successfully with sapropterin monotherapy have required dietary treatment in long-term follow-up. We aimed to determine the baseline factors associated with future initiation of dietary treatment in these patients. METHODS: Data were obtained retrospectively from the medical records of 80 PKU patients started on sapropterin monotherapy before 3 months of age between 2011 and 2021. RESULTS: The patients were followed for a median of 3.9 years (Q1-Q3: 2.5-5.75 years). Sapropterin was tapered down and discontinued in 5 patients (6.3%) as their Phe levels remained below 360 µmol/L without treatment. Sapropterin monotherapy was sufficient in 62 patients (77.5%), while 13 (16.2%) required dietary treatment. Phe and tyrosine (Tyr) levels, and Phe:Tyr ratios differed significantly among the patients maintained on sapropterin monotherapy and those started on dietary treatment, but the Phe:Tyr ratio at diagnosis was the most important independent baseline variable (OR: 1.61, 95% CI: 1.15-2.27, p = 0.006), with Phe:Tyr ratio at diagnosis >5.25 associated with dietary treatment (sensitivity: 90.0%, specificity: 81.8%). Genotypic phenotype value (GPV), unavailable at baseline, was also associated with dietary treatment (median GPV 9.2 vs. 3.8, p = 0.006), but some genotypes were not specific to the final treatment modality. DISCUSSION: We propose that the Phe:Tyr ratio at diagnosis is an important indicator to predict dietary requirement in young infants initially managed with sapropterin monotherapy.
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Fenilalanina Hidroxilasa , Fenilcetonurias , Humanos , Lactante , Estudios Retrospectivos , Calidad de Vida , Fenilalanina , Fenilcetonurias/tratamiento farmacológico , Fenilcetonurias/genética , Dieta , Biopterinas , Fenilalanina Hidroxilasa/genéticaRESUMEN
This study aims to evaluate the effect of synchronization and different superstimulation protocols on oocyte yield before ovum-pick up (OPU) to provide a homogeneous follicle population. Animals in all study groups except the control group applied a synchronization protocol including modified ovsynch+progesterone and dominant follicle ablation (DFA, on the 6th day following synchronization). In group 1, oocytes were retrieved by ultrasonography only on the 4th day after DFA. In group 2, 250 µg pFSH (100 µg IM, 150 µg SC) was administered as a single dose on the 2nd day following DFA, and oocytes were retrieved on the 2nd day following this injection. In group 3, 250 µg pFSH was administered IM in four equal doses 12 h apart on the 1st and 2nd days following DFA, and oocytes were retrieved 2 days after the last FSH injection. In group 4, 250 µg pFSH (IM) dissolved in Montanide ISA 206 adjuvant was administered as a single dose on the 2nd day following DFA, and oocytes were retrieved 2 days after this treatment. In the control group (group 5), oocytes were retrieved from animals without any hormonal treatment on a random day of the oestrus cycle. In order to assess the follicle population in the ovary on the day of OPU, the number of follicles according to their diameter was determined by ultrasonography in all groups. The ratio of medium-sized follicles (3-8 mm) was higher in the synchronized groups (Groups 1, 2, 3 and 4) than in the control group (Group 5) (p < .05). It was determined that the total number of oocytes obtained after OPU and the number of oocytes of suitable quality (Grade A and B) in in vitro embryo production were higher in the superstimulated groups (Groups 2, 3 and 4) compared to the control group. It was found that the number of Grade-A quality oocytes was higher in the superstimulated groups (Groups 2, 3 and 4) than the other groups. As a result, it was found that the synchronization and superstimulation treatments prior to the OPU increased the ratio of medium-sized follicles and the total number of oocytes obtained. In addition to the synchronization protocol, it was determined that superstimulation treatments increased the oocyte quality obtained with OPU. Furthermore, it was observed that a single dose of FSH dissolved in Montanide ISA 206 adjuvant produced a superstimulation response similar to that produced by repeated doses of FSH.
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Fertilización In Vitro , Hormona Folículo Estimulante , Femenino , Animales , Hormona Folículo Estimulante/farmacología , Fertilización In Vitro/métodos , Fertilización In Vitro/veterinaria , Inducción de la Ovulación/veterinaria , Inducción de la Ovulación/métodos , Oocitos/fisiología , ÓvuloRESUMEN
To investigate the relationship between apical periodontitis [AP] severity and inflammatory markers [interleukin (IL)-12, tumor necrosis factor-alpha (TNF-α), and Mid-Regional Pro Adrenomedullin (MR-proADM)] in patients with AP. A total of 162 subjects were divided into three categories: AP group (n = 80), periodontitis (P) group (n = 42), healthy control group (n = 40). The scoring of disease severity in 80 AP patients without any periodontal disease, using dental radiographs, was based on "The Abscess Score" (AS), as those having at least 1 tooth with AP and severity of PAI 3-4 were classified as AS 1 (mild); those with only1 tooth and severity of PAI 5 as AS 2 (moderate) and those having > 2 tooth with severity of PAI 5 as AS 3 (severe). Blood samples were collected from all of the patients. Enzyme-linked immunosorbent assay was used to evaluate the samples. The MR-pro ADM levels of both the AP and P groups were considerably higher than the control group (p < 0.01). The IL-12 levels of the AP group were higher than the P and control groups (p < 0.05). TNF-α levels of the P group were significantly higher than both the AP and control groups (p < 0.01). MR-pro ADM levels of both the AP and Periodontitis groups were higher than the control group. TNF-α was a biomarker of periodontitis, while IL-12 was a biomarker of apical periodontitis.
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Periodontitis Periapical , Periodontitis , Humanos , Factor de Necrosis Tumoral alfa/metabolismo , Regulación hacia Arriba , Adrenomedulina , Biomarcadores , Interleucina-12RESUMEN
Intercellular Adhesion Molecule-1 (ICAM-1) is considered to be a cancer biomarker in the assessment of metastatic potential in patients and an early indicator of atherosclerosis. A labelless biosensor based on the surface plasmon resonance (SPR) signal from the specific affinity interaction of an aptamer and a soluble ICAM-1 protein was developed for blood samples. The developed aptasensor provided real-time information on the concentration of the ICAM-1 protein in blood when integrated to a purification step based on a magnetic pull-down separation. The SPR aptasensor was highly specific with a limit of detection of 1.4/0.2 ng ml-1, which was achieved through aptamer-functionalized silica-coated magnetic nanoparticles.
Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , Oro , Humanos , Molécula 1 de Adhesión Intercelular , Límite de Detección , Resonancia por Plasmón de SuperficieRESUMEN
The purpose of this study is to investigate the effects of different root canal irrigation protocols applied to the dentin and artificial aging procedures on the micro pushout bond strength (mPBS) between dentin and hybrid ceramic posts. Seventy-five single-rooted mandibular premolar teeth were divided into 5 groups (Gr1-5). 50 of the teeth were used for the mPBS tests (n = 10), whereas 25 were used for the smear layer examinations (n = 5). Post space were prepared and irrigated with different irrigation-protocols in each group. (Gr1:[SS], Gr2:[NaOCl] + SS, Gr3:[EDTA] + NaOCl + SS, Gr4:[MA] + NaOCl + SS, Gr5:[Ch] + NaOCl + SS). Post and core pattern were fabricated with pattern resin and a fiber post, after scanning, the posts were milled with Vita Enamic resin ceramic block, and cemented. After 7 days the roots were sliced at thicknesses of 1 mm; half of them were subjected to mPBS test, while the other half were tested after undergoing mechanical cycling for artificial aging. For data analysis, the Shapiro-Wilk test was utilized to test normal distributions, 3-way analysis of variance was used to compare mPBS, and Tukey's HSD test was conducted for multiple comparisons. SEM analysis was performed for examination of failure modes and smear layer removal. Different root canal irrigation protocols affected mPBS significantly. While Gr4 had the highest mPBS, Gr1 had the lowest. Regarding to different zones, the highest mPBS was in coronal zone, and the lowest one was in the apical zone. The aging procedure also led to a statistically-significant decrease in mPBS. Most frequent failure modes were cohesive failure in dentin and mixed failure. Irrigation with 7%MA (Gr4) showed better performance than 17% EDTA (Gr3) in smear layer removal, especially at the apical zone of the tooth. This is critical for the success of root canal treatment and increased the mPBS to a higher extent in all zones of the tooth.
Asunto(s)
Recubrimiento Dental Adhesivo , Capa de Barro Dentinario , Humanos , Irrigantes del Conducto Radicular/uso terapéutico , Irrigantes del Conducto Radicular/química , Cavidad Pulpar , Dentina , Recubrimiento Dental Adhesivo/métodos , Hipoclorito de Sodio/uso terapéutico , Hipoclorito de Sodio/química , Ácido Edético/análisis , Ácido Edético/química , Ácido Edético/farmacología , Diente Premolar , Preparación del Conducto Radicular/métodos , Ensayo de MaterialesRESUMEN
Disorders of intracellular trafficking are a group of inherited disorders, which often display multisystem phenotypes. Vacuolar protein sorting (VPS) subunit C, composed of VPS11, VPS18, VPS16, and VPS33A proteins, is involved in tethering of endosomes, lysosomes, and autophagosomes. Our group and others have previously described patients with a specific homozygous missense VPS33A variant, exhibiting a storage disease phenotype resembling mucopolysaccharidosis (MPS), termed "MPS-plus syndrome." Here, we report two siblings from a consanguineous Turkish-Arabic family, who have overlapping features of MPS and intracellular trafficking disorders, including short stature, coarse facies, developmental delay, peripheral neuropathy, splenomegaly, spondylar dysplasia, congenital neutropenia, and high-normal glycosaminoglycan excretion. Whole exome sequencing and familial segregation analyses led to the homozygous NM_022575.3:c.540G>T; p.Trp180Cys variant in VPS16 in both siblings. Multiple bioinformatic methods supported the pathogenicity of this variant. Different monoallelic null VPS16 variants and a homozygous missense VPS16 variant had been previously associated with dystonia. A biallelic intronic, probably splice-altering variant in VPS16, causing an MPS-plus syndrome-like disease has been very recently reported in two individuals. The siblings presented herein display no dystonia, but have features of a multisystem storage disorder, representing a novel MPS-plus syndrome-like disease, associated for the first time with VPS16 missense variants.
Asunto(s)
Mucopolisacaridosis/genética , Mutación Missense , Proteínas de Transporte Vesicular/genética , Anomalías Múltiples , Femenino , Homocigoto , Humanos , Lactante , Masculino , Mucopolisacaridosis/patología , Linaje , Fenotipo , Hermanos , SíndromeRESUMEN
We report a 42-year-old patient who had Hodgkin lymphoma and developed bilateral symmetrical peripheral gangrene (SPG) in the feet and hands, which occurred during septic shock after autologous hematopoietic stem-cell transplantation. SPG is a rare but severe complication of disseminated intravascular coagulation (DIC) and is frequently associated with sepsis. The pathophysiology of SPG includes DIC-mediated intravascular thrombosis and thrombotic occlusion of microcirculation, resulting in low blood flow. Sepsis-induced hypotension has been suspected as one of the other causes of SPG, and it is thought to be aggravated by vasopressor treatments given for hypotension. Our patient first experienced coldness, paleness, and cyanosis in his body's acral parts, and then SPG later developed in both his feet and hands. Septic shock management was performed with cytokine hemoadsorption, broad-spectrum antibiotics, and massive fluid replacement rapidly. The patient fully recovered without the need for amputation. Hemoadsorption is an extracorporeal cytokine-adsorption method for removing excess cytokines. Prompt management of septic shock and early monitoring of peripheral ischemia are essential to avoid SPG.
Asunto(s)
Gangrena/etiología , Gangrena/terapia , Enfermedad de Hodgkin/complicaciones , Enfermedad de Hodgkin/diagnóstico , Choque Séptico/terapia , Adulto , Antibacterianos/farmacología , Citocinas/metabolismo , Progresión de la Enfermedad , Coagulación Intravascular Diseminada , Hemabsorción , Humanos , Hipotensión , Masculino , Sepsis/complicaciones , Sepsis/fisiopatología , Trombocitopenia , Resultado del Tratamiento , Vasoconstrictores/efectos adversosRESUMEN
In addition to tetrahydrobiopterin deficiencies and phenylalanine hydroxylase deficiency (phenylketonuria) due to PAH variants, the deficiency of the co-chaperone protein DNAJC12 was identified in 2017 as a novel cause of inherited hyperphenylalaninemia, revealing the genetic etiology in previously unresolved cases. In this study, we aimed to investigate DNAJC12 deficiency in non-tetrahydrobiopterin-deficient persistent hyperphenylalaninemia cases without biallelic PAH variants in a single pediatric metabolic center. It was determined retrospectively that 471 patients with non-tetrahydrobiopterin deficiency-hyperphenylalaninemia had undergone PAH gene sequencing and 451 patients had biallelic variants in PAH. DNAJC12 sequencing was performed in the remaining 20 patients, identifying a previously reported homozygous splice-site variant (c.158-2A > T) in one patient with axial hypotonia and developmental delay, and a novel, homozygous c.404del (p.Arg135Lysfs*21) frameshift variant in an asymptomatic patient. In segregation analysis, the asymptomatic patient's both parents were also found to be homozygous for this variant and hyperphenylalaninemic. The parents may have had academic difficulties but intellectual disability could not be confirmed due to lack of cooperation. The symptomatic patient significantly benefited from treatment with sapropterin dihydrochloride and neurotransmitter precursors. DNAJC12 deficiency might be responsible for approximately 10% or more of cases with unexplained hyperphenylalaninemia. The phenotypic spectrum is broad, ranging from early infantile hypotonia to incidental diagnosis in adulthood. Similar to tetrahydrobiopterin deficiencies, early diagnosis and treatment with sapropterin dihydrochloride and neurotransmitter precursors can be beneficial, supporting the analysis of DNACJ12 gene in patients with unexplained hyperphenylalaninemia.
Asunto(s)
Errores Innatos del Metabolismo de los Aminoácidos/genética , Proteínas del Choque Térmico HSP40/deficiencia , Fenilalanina/sangre , Errores Innatos del Metabolismo de los Aminoácidos/complicaciones , Biopterinas/análogos & derivados , Biopterinas/uso terapéutico , Niño , Discapacidades del Desarrollo/genética , Femenino , Variación Genética , Humanos , Recién Nacido , Discapacidad Intelectual/genética , Masculino , Hipotonía Muscular/genética , Neurotransmisores/uso terapéutico , Fenilalanina Hidroxilasa/genética , Isoformas de Proteínas/genéticaRESUMEN
A Correction to this paper has been published: https://doi.org/10.1007/s11011-021-00759-8.
RESUMEN
To examine the use of IL-6 and PAPP-A along with the new imaging for scoring methods to determine the severity of apical periodontitis (AP). The scoring of disease severity in 121 patients with AP, using dental radiographs, was based on "The Apical Periodontitis Grading Scale" (APGS), as those having 1 tooth with AP and severity of 3-4 were classified as grade 1 (mild); those having > 1 tooth and severity of 3-4 as grade 2 (moderate) and those with at least one tooth with a severity of 5 as grade 3 (severe). 45 systematically and orally healthy volunteers were selected as the control group (grade 0). Dental demographic data of all participants were recorded. Besides, venous blood samples were collected to study their complete blood count and levels of IL-6 and PAPP-A. Both NLR and PAPP-A levels of the grade 3 AP patients were found significantly higher than those of the control group and grade 1 and grade 2 AP patients (2.55 ± 1.40 vs 1.98 ± 1.05, 1.94 ± 0.58 and 1.86 ± 0.50; 0.606 ± 0.211 vs 0.422 ± 0.273, 0.447 ± 0.224 and 0.436 ± 0.242, p < 0.01, respectively). AP grades identified by the new scoring system were moderately correlated with IL-6 levels (Spearman r = 0.4168, p < 0.001). The risk of patients with IL-6 levels above 12.5 pg/ml to develop AP (relative risk) was found to be 2.19 times higher than that in those with lower IL-6 levels (98% Cl 1.233-3.905, p < 0.0024). APGS classification suggested to determine AP severity was found to be significantly correlated with levels of inflammatory markers IL-6 and PAPP-A.
Asunto(s)
Interleucina-6 , Periodontitis Periapical , Biomarcadores , Femenino , Humanos , Periodontitis Periapical/diagnóstico por imagen , Embarazo , Proteína Plasmática A Asociada al EmbarazoRESUMEN
The present study was aimed to investigate the effects of double-dose gonadotropin-releasing hormone (GnRH) injection on the induction of oestrus and some reproductive performance parameters in Awassi ewes during the non-breeding season. In the study, 100 ewes were treated with a vaginal sponge containing 60 mg medroxyprogesterone acetate for 7 days in the anoestrus (day 0). PMSG 500 IU and 250 µg cloprostenol sodium were injected on the day of removal of the sponge (day 7). Ewes in Group 1 (n = 31) were not subjected to any hormonal treatment. Ewes in Group 2 (n = 31) were given 50 µg GnRH 48th hour after removal of the sponge. Ewes in Group 3 (n = 33) were given 50 µg GnRH 48th hour after the removal of the sponge and 50 µg GnRH 12th day after post-mating. The results obtained in the study showed that there were no statistical differences between the Groups 1, 2 and 3 in terms of oestrus rates (82.8%, 68.9%, 72.7%), conception rates (66.7%, 55.0%, 54.2%), multiple pregnancy rates (28.5%, 50.0%, 30.7%) and litter sizes (1.28, 1.50, 1.31). No significant increases in P4 concentration were observed in Group 3 treated with GnRH at the 12th day after post-mating; however, a numerically lower (p > 0.05) late embryonic-early fetal mortality rate was observed in Group 3 (0%), when compared with the values obtained in Group 1 (12.5%) and Group 2 (9.1%). In conclusion, after short-term progestagen administration during the non-breeding season, double-dose GnRH injections did not increase P4 concentration and had no significant differences on reproductive performance parameters among groups.